CN107938370B

CN107938370B - Bacteria cellulose film composite material and preparation method with nanometer spider web structure

Info

Publication number: CN107938370B
Application number: CN201711084713.0A
Authority: CN
Inventors: 陈仕艳; 陈燕; 李喆; 王华平; 盛楠; 魏佳欣; 李文颖
Original assignee: Donghua University
Current assignee: Donghua University
Priority date: 2017-11-07
Filing date: 2017-11-07
Publication date: 2019-06-21
Anticipated expiration: 2037-11-07
Also published as: CN107938370A

Abstract

The present invention relates to a kind of bacteria cellulose film composite material and preparation method with nanometer spider web structure, methods are as follows: it is 10 that the bacterial strain that isoelectric point is 3.5~4.0, which is dispersed in the strain cultured solution that pH value is 4.5~5.0, and obtains strain cell density⁹~10¹²The culture bacterium solution of a/mL, then the stationary culture on the non-woven fabrics of surface static water contact angle≤70 ° is added dropwise in culture bacterium solution, composite material is made.The adjacent two layers of the composite material are respectively the non-woven fabrics of bacteria cellulose film and surface hydrophilic, bacteria cellulose film is mainly made of trunk bacteria cellulose fibre and mycobacterium cellulose fibre, be cross-linked with each other the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film between trunk bacteria cellulose fibre itself, the networking between trunk bacteria cellulose fibre of mycobacterium cellulose fibre.The method of the present invention is simple and easy, and composite material obtained has many advantages, such as good biocompatibility, good permeability and high-strength light.

Description

Bacteria cellulose film composite material and preparation method with nanometer spider web structure

Technical field

The invention belongs to field of compound material, and in particular to the bacteria cellulose film composite material with nanometer spider web structure And preparation method thereof.

Background technique

Destruction due to industrial fast development and the mankind to natural environment, air pollution are got worse, in recent years, I State saves more there is lasting fog weather, has seriously affected breathing and the lung health of people.And main " gray haze arch-criminal " It is fine particle (PM2.5) of the partial size less than 2.5 μm, also known as lung particulate matter in air.On the other hand, pass through respiratory tract The various diseases propagated also threaten the health of the mankind, such as influenza and bird flu etc..Just because of atmosphere pollution exacerbation with The burst of disease is infected, properly protecting in daily life is particularly important, therefore the exploitation of advanced composite material (ACM) is always It is a hot spot of scientific research.

For composite material, the composite material that only effective aperture is less than the size of filtrate, which is just able to achieve, effectively to be hindered Every the effect of protection.And the particle size distribution of PM2.5 is substantially that 100~700nm of diameter accounts for 31.8%, 800~2500nm Account for 58.7%, be 70~150nm by the viral diameters of droplet transmission, wherein influenza virus diameter is 80~120nm, SARS Viral diameters are about 70~80nm, and coronavirus diameter is 80~150nm, and A (H 1 N 1) virus diameter is 80~120nm.Bacterium Diameter than virus it is big, the diameter of most bacteriums is between 500~5000nm.Therefore for composite material, work as hole When diameter is less than 70nm, lung particulate matter and virus can be effectively intercepted.

The composite material that above two hazardous material can effectively be obstructed at present is mainly melt-blown micro fibre and nanofiber material Expect (such as patent CN 102115956 A and patent CN101564914 B).For the former, due to melt-spraying spinning forming process with Traditional spinning technique is entirely different, not can be carried out effective control to the drawing-off of fiber, so the thickness of fiber obtained is equal Evenness is poor, and the degree of orientation of fiber is lower, and strength is insufficient, in addition, the power consumption for preparing such fiber is also larger, cost is inclined Height, and for the latter, method of electrostatic spinning is to prepare one of most important method of nanofiber at present, but it is limited by high energy consumption With the technical bottleneck of low-yield, but also nano-fiber material cannot achieve the large-scale production of high efficiency, low cost.

Bacteria cellulose be synthesize by microorganisms such as acetobacter xylinums by metabolism sugar source, vivo biodistribution and external stratification oneself The nano-fiber material of obtained high-crystallinity and high-purity is assembled, which has hyperfine network structure. In addition, it is received significant attention with having the characteristics that raw material is natural, synthesis process is mild efficient and final product is environmental-friendly, it is One of ten big hitech materials of 21 century most prospect, and the preparation that biological synthesis process is also current most industrial prospect is received A kind of method of rice fiber, while it is as natural nanofiber has more large specific surface area, hole, good permeability and gently The advantages that matter is high-strength can be fully applicable to the protection field such as filter and adsorb.But the bacteria cellulose grown naturally, its inside Fiber alignment is unordered, while its effective aperture can not meet the size requirement of lung particulate matter and virus.

Therefore, develop a kind of lung particulate matter that can effectively obstruct in air and disinfect pathogen has industrial metaplasia again The bacteria cellulose film composite material of antenatal scape has great realistic meaning.

Summary of the invention

It is complicated the purpose of the present invention is to solve nanofiber preparation process existing in the prior art and obtained receive Rice fiber the problem of cannot effectively obstructing the lung particulate matter and disinfect pathogen in air, provide a kind of simplicity has work Industry prospect of production and the bacteria cellulose film composite material that can effectively obstruct the lung particulate matter and disinfect pathogen in air Preparation method.Composite material of the invention has effectively interception effect to lung particulate matter and disinfect pathogen, can be more The deficiencies in the prior art or defect are mended, production and living needs are met, in addition, present invention process is simple, it is easy to operate, prepare skill Art is controllable, pollution-free, low in cost, can prepare various bacteria cellulose membrane composite material, the composite material of preparation as needed It can be applied to the protection field such as adsorb and filter, there is very wide industrial applications prospect.

In order to achieve the above object, the technical solution adopted by the present invention are as follows:

Bacteria cellulose film composite material with nanometer spider web structure, with two layers or more of structure, wherein adjacent two Layer is respectively the non-woven fabrics of bacteria cellulose film and surface hydrophilic with nanometer spider web structure；

The bacteria cellulose film with nanometer spider web structure is mainly by trunk bacteria cellulose fibre and mycobacterium Cellulose fibre is constituted；

The trunk bacteria cellulose fibre is to be cross-linked with each other to form three-dimensional mesh structure and constitute bacterium fibre between itself Tie up the bacteria cellulose fibre of the skeleton of plain film；

The mycobacterium cellulose fibre is the bacteria cellulose fibre into the net between trunk bacteria cellulose fibre.

As a preferred technical scheme:

Bacteria cellulose film composite material as described above with nanometer spider web structure, the trunk bacteria cellulose are fine Dimension passes through β -1,4- glucosides key connection with mycobacterium cellulose fibre.

Existing melt-blown micro fibre average diameter is 0.5~5 μm, and fiber is thicker, and during spinning technique, made The fiber thickness uniformity obtained is poor, and the degree of orientation is lower, and strength is insufficient, in addition, the power consumption for preparing fiber is also larger, cost It is higher.Although and electrostatic network technology preparation nanofiber can also obtain this nanometer spider web structure, what is frequently resulted in is Nanometer spider web and nanofiber compound two-dimensional nano network structure are simple physics between nanometer spider web and nanofiber Accumulation, therefore the mechanical properties decrease compared with simple nanofiber, while it being limited by high energy consumption, the technical bottleneck of low-yield, So that nano-fiber material cannot achieve the large-scale production of high efficiency, low cost.And nanometer spider web structure of the invention is by trunk The three-dimensional mesh structure that bacteria cellulose fibre and mycobacterium cellulose fibre are constituted, and trunk fiber and branched fiber are Bacterial strain biochemistry synthesizes to obtain, and is the entirety being formed by connecting by chemical bond between fiber and fiber.Chemically form from the point of view of, be all by Glucopyranose monomer passes through polymer made of β -1,4- glucosides key connection.Therefore, nanometer spider web structure of the invention is thin Fungin film has excellent mechanical property, meanwhile, bacteria cellulose film of the invention is made by biological synthesis process, operation letter Single, green non-pollution, low energy consumption, has wide industrial prospect.

Bacteria cellulose film composite material as described above with nanometer spider web structure, the mesh hole of the nanometer spider web Diameter is 20~300nm；The bacteria cellulose film with nanometer spider web structure with a thickness of 0.1~9mm, density is 5~ 8mg/cm³；The average diameter of the trunk bacteria cellulose fibre is 30~80nm；The mycobacterium cellulose fibre is put down Equal diameter is 5~20nm.

Bacteria cellulose film composite material as described above with nanometer spider web structure, the non-woven fabrics of the surface hydrophilic With a thickness of 0.1~1mm, surface density is 15~40g/m², handled and be made through surface hydrophilic by non-woven fabrics；

The surface hydrophilic refers to Static water contact angles≤70 ° of nonwoven surface.

Bacteria cellulose film composite material as described above with nanometer spider web structure, the non-woven fabrics are by averagely straight The fiber that diameter is 20~50 μm is constituted, and material is polypropylene (PP), polyethylene terephthalate (PET), polyamide (PA), viscose glue, polyacrylonitrile (PAN), polyglycolide, polylactic acid (PLA), polyethylene (PE), polylactide glycolate copolymer (PLGA) or chitosan；

The surface hydrophilic processing, which refers to, is grafted on non-woven fabrics material surface for negatively charged hydrophilic radical.

The present invention also provides preparations to have the bacteria cellulose film composite material of nanometer spider web structure as described above The bacterial strain that isoelectric point is 3.5~4.0, is dispersed in the strain cultured solution that pH value is 4.5~5.0 that obtain bacterial strain thin by method Born of the same parents' density is 10⁹~10¹²The culture bacterium solution of a/mL, then be added dropwise bacterium solution is cultivated on the surface of surface static water contact angle≤70 ° The bacteria cellulose film composite material with nanometer spider web structure is made in stationary culture on hydrophilic non-woven fabrics.

In the biochemical synthesis process of BC, strain cell is urged as biosynthesis reaction device, by glucose small molecule in enzyme Change effect is lower to generate cellulose microfibril (Cellulose microfibrils), diameter 1.78nm, and by cell wall side Aperture squeezes out.Since, there are multiple such apertures, adjacent cellulose microfibril is in hydrogen bond on each strain cell wall Under the action of formed microfibril bundle (Bundle), diameter be 3~4nm.These microfibril bundles further link just formation diameter The fiber filament band (Ribbon) of 30~100nm.Strain cell carries microfibril bundle or fiber filament band is randomly transported in culture solution Dynamic, microfibril bundle or fiber filament band are interleaved with each other to form irregular netted or flocculent structure.Strain cultured solution pH in the present invention Value is 4.5~5.0, and bacterial strain isoelectric point is 3.5~4.0, therefore strain cell is negatively charged in culture bacterium solution, simultaneously as Nonwoven surface used is grafted negatively charged hydrophilic radical, and under identical charges effect, strain cell is in culture bacterium solution Movement rate accelerate, strain cell carries the quick random motion in strain culturing bacterium solution of microfibril bundle or fiber filament band, Microfibril bundle entrained by strain cell or fiber filament band intertwine with each other and combine under hydrogen bond action, while with identical charges bacterium Strain cell is mutually exclusive, is finally formed under the collective effect of hydrogen bond gravitation and charge repulsion and is similar to the netted of spider web frame Fibrous material.This reticular structure is (a small amount of by trunk bacteria cellulose fibre (fiber filament band) and mycobacterium cellulose fibre Microfibril bundle is by Hydrogenbond) it constitutes.

As a preferred technical scheme:

Method as described above, specifically includes the following steps:

1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, after sterilized processing It is laid in culture vessel；

2) fermentation culture is prepared: by mass percentage, unit wt%, each component and its content in fermentation culture It is as follows:

3) activate: by the isoelectric point of refrigeration be 3.5~4.0 bacterial strain in thaw at RT, be placed in 100~250mL fermentation Carrying out shaking table culture in culture solution, the revolving speed of shaking table culture is 50~150r/min, and temperature is 28~32 DEG C, the time is 6~ 24h；

4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:2~20, at 28~32 DEG C Lower stationary culture 5~9 days；

5) be centrifuged: strain cell cell number after overactivation and amplification increases again at geometry, under the action of the centrifugal force bacterium Bacterium solution centrifugation removal supernatant after amplification is obtained strain cell precipitating and mixed with a small amount of culture solution by strain cell forming precipitating Concentration bacterium solution；

6) prepare strain cultured solution: it is 4.5~5.0 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains bacterial strain Culture solution；

7) it cultivates: strain cultured solution is added in concentration bacterium solution, and evenly dispersed, obtaining strain cell density is 10⁹~ 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise on the non-woven fabrics of surface static water contact angle≤70 °, 28~ Stationary culture 3~9 days at 32 DEG C obtain the bacteria cellulose film composite material with nanometer spider web structure.Culture of the invention Bacterium solution is high density bacterium solution, and the high density bacterium solution refers to that strain cell is dispersed in obtained mixing in strain cultured solution Object, wherein strain cell density is 109~1012/ml, micro- entrained by strain cell with the increase of strain cell density Fiber tow or fiber filament band intertwine with each other probability raising, advantageously form the lesser spider web structure of nanometer mesh aperture.

Method as described above, the bacterial strain that the isoelectric point is 3.5~4.0 are selected from gluconacetobacter, produce acetobacter, acetify Bacillus, Pasteur's acetobacter, grape saccharobacillus, Agrobacterium, rhizobium, sarcine, Pseudomonas cepacia, Pseudomonas cocovenenans or One or more of campylobacter jejuni.Bacterial strain in the present invention includes but is not limited to this, and isoelectric point is 3.5~4 and being capable of biology The microorganism of synthesis bacterial fibers film can be used as bacterial strain of the invention, only enumerate some common microorganisms here.

Method as described above, the sterilization treatment refer to high pressure steam sterilization, ultraviolet irradiation sterilizing, ethylene oxide sterilizing Or Co 60-radiated by gamma-ray sterilizing；The centrifugal rotational speed of the centrifugation be 2000~8000r/min, centrifugation time be 3~ 10min；It is described it is evenly dispersed refer to using eddy blending machine, 10~30s of continuous shaking at 500~2000rpm.Centrifugal force mistake It is small, it is not easily formed precipitating, centrifugal force crosses senior general and causes to damage to strain cell, therefore revolving speed need to be controlled in centrifugal treating With the time in zone of reasonableness.

Invention mechanism:

Strain cultured solution pH value is 4.5~5.0 in the present invention, and bacterial strain isoelectric point is 3.5~4.0, the culture of strain cell Liquid pH value is greater than isoelectric point, thus strain cell is negatively charged in culture bacterium solution, simultaneously as nonwoven surface used connects The negatively charged hydrophilic radical of branch, under identical charges effect, movement rate of the strain cell in culture bacterium solution is accelerated, bacterial strain Cell carries the quick random motion in strain culturing bacterium solution of microfibril bundle or fiber filament band, fento entrained by strain cell Tow or fiber filament band intertwine with each other and combine under hydrogen bond action, while mutually exclusive with identical charges strain cell, finally The reticular fibre material for being similar to spider web frame is formed under the collective effect of hydrogen bond gravitation and charge repulsion.

The utility model has the advantages that

(1) preparation method of the bacteria cellulose film composite material with nanometer spider web structure of the invention, it is simple and easy, Easy to operate, green non-pollution has fabulous promotional value；

(2) the bacteria cellulose film composite material with nanometer spider web structure of the invention has good biocompatibility, ratio Surface area is big, hole is more, good permeability and the advantages that high-strength light, has huge application the protection field such as filtering and adsorbing Potentiality.

Detailed description of the invention

Fig. 1 is bacteria cellulose film scanning electron microscope schematic diagram prepared by the embodiment of the present invention 1；

Fig. 2 is bacteria cellulose film scanning electron microscope schematic diagram prepared by the embodiment of the present invention 2；

Fig. 3 is bacteria cellulose film scanning electron microscope schematic diagram prepared by the embodiment of the present invention 3.

Specific embodiment

The invention will be further elucidated with reference to specific embodiments.It should be understood that these embodiments are merely to illustrate this hair It is bright rather than limit the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, art technology Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited Fixed range.

Embodiment 1

A kind of preparation method of the bacteria cellulose film composite material with nanometer spider web structure, steps are as follows:

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through high steam It is laid in culture vessel after bacterium processing, wherein the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, table Face hydrophilic treated, which refers to, is grafted on the non-woven fabrics being made of PP material that average diameter is 30 μm for negatively charged hydrophilic radical Surface, the non-woven fabrics of surface hydrophilic with a thickness of 0.1mm, surface density 20g/m², surface static water contact angle is 70 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: glucose 2, peptone 0.05, yeast extract 0.05, citric acid 0.01, disodium hydrogen phosphate 0.02, potassium dihydrogen phosphate 0.01, surplus is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 gluconacetobacter in thaw at RT, be placed in 100mL hair Shaking table culture is carried out in ferment culture solution, the revolving speed of shaking table culture is 50/min, and temperature is 28 DEG C, time 6h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:2, being stood at 28 DEG C Culture 5 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 2000r/ Min, centrifugation time 3min；

(6) prepare strain cultured solution: it is 4.5 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 500 rpm continuous vibration Dynamic 10s, obtaining strain cell density is 10⁹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 70 °, stationary culture 3 days at 28 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure of preparation is double-layer structure, respectively has nanometer The bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic have the micro- of the bacteria cellulose film of nanometer spider web structure Tissue as shown in Figure 1, the trunk bacteria cellulose fibre and average diameter for being mainly 30nm by average diameter be 5nm branch it is thin Fungin fiber is constituted, and trunk bacteria cellulose fibre is to be cross-linked with each other to form three-dimensional mesh structure and constitute thin between itself The bacteria cellulose fibre of the skeleton of fungin film；Mycobacterium cellulose fibre is between trunk bacteria cellulose fibre Bacteria cellulose fibre into the net.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre are connected by β -1,4- glycosidic bond It connects.Nanometer spider web mesh aperture be 160~300nm, with nanometer spider web structure bacteria cellulose film with a thickness of 0.1mm, density 5mg/cm³。

Embodiment 2

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through ultraviolet irradiation It is laid in culture vessel after bacterium processing, wherein the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, table Face hydrophilic treated, which refers to, is grafted on the non-woven fabrics being made of PET material that average diameter is 50 μm for negatively charged hydrophilic radical Surface, the non-woven fabrics of surface hydrophilic with a thickness of 0.2mm, surface density 30g/m², surface static water contact angle is 69 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: sucrose 3, peptone 0.09, yeast extract 0.08, citric acid 0.02, disodium hydrogen phosphate 0.08, potassium dihydrogen phosphate 0.06, Surplus is water；

(3) it activates: being 3.5~4.0 to acetify bacillus in thaw at RT by the isoelectric point of refrigeration, be placed in 200mL fermentation Shaking table culture is carried out in culture solution, the revolving speed of shaking table culture is 80r/min, and temperature is 29 DEG C, time 10h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:10, being stood at 29 DEG C Culture 6 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 3000r/ Min, centrifugation time 4min；

(6) prepare strain cultured solution: it is 4.8 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 800 rpm continuous vibration Dynamic 15s, obtaining strain cell density is 10¹¹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 69 °, stationary culture 8 days at 30 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure of preparation is double-layer structure, respectively has nanometer The bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic have the micro- of the bacteria cellulose film of nanometer spider web structure Tissue is as shown in Fig. 2, the branch that the trunk bacteria cellulose fibre and average diameter for being mainly 72nm by average diameter are 18nm Bacteria cellulose fibre is constituted, and trunk bacteria cellulose fibre is to be cross-linked with each other to form three-dimensional mesh structure and constitute between itself The bacteria cellulose fibre of the skeleton of bacteria cellulose film；Mycobacterium cellulose fibre be trunk bacteria cellulose fibre it Between bacteria cellulose fibre into the net.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glycosidic bond Connection.The mesh aperture of nanometer spider web is 50~70nm, the bacteria cellulose film with nanometer spider web structure with a thickness of 8mm, Density is 7.5mg/cm³。

Embodiment 3

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, ethane via epoxyethane goes out It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the nonwoven surface being made of PA material that average diameter is 40 μm for negatively charged hydrophilic radical, The non-woven fabrics of surface hydrophilic with a thickness of 0.3mm, surface density 18g/m², surface static water contact angle is 62 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: glucose 2, peptone 0.5, yeast extract 0.08, citric acid 0.09, disodium hydrogen phosphate 0.1, potassium dihydrogen phosphate 0.03, Surplus is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 grape saccharobacillus in thaw at RT, be placed in 200mL hair Shaking table culture is carried out in ferment culture solution, the revolving speed of shaking table culture is 120r/min, and temperature is 31 DEG C, time 12h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:15, being stood at 30 DEG C Culture 5~9 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 2000r/ Min, centrifugation time 10min；

(6) prepare strain cultured solution: it is 4.6 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, continuously shakes at 1000rpm Dynamic 10s, obtaining strain cell density is 10¹¹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 62 °, stationary culture 4 days at 29 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure of preparation is double-layer structure, respectively has nanometer The bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic have the micro- of the bacteria cellulose film of nanometer spider web structure Tissue is as shown in figure 3, the branch that the trunk bacteria cellulose fibre and average diameter for being mainly 50nm by average diameter are 12nm Bacteria cellulose fibre is constituted, and trunk bacteria cellulose fibre is to be cross-linked with each other to form three-dimensional mesh structure and constitute between itself The bacteria cellulose fibre of the skeleton of bacteria cellulose film；Mycobacterium cellulose fibre be trunk bacteria cellulose fibre it Between bacteria cellulose fibre into the net.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glycosidic bond Connection.Nanometer spider web mesh aperture be 150~300nm, with nanometer spider web structure bacteria cellulose film with a thickness of 3mm, density 6mg/cm³。

Embodiment 4

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, through Co 60-gamma It is laid in culture vessel after x ray irradiation x sterilization treatment, the non-woven fabrics of surface hydrophilic is to be handled to make through surface hydrophilic by non-woven fabrics , surface hydrophilic processing, which refers to negatively charged hydrophilic radical is grafted on average diameter to be 20 μm, to be made of viscose glue material Nonwoven surface, the non-woven fabrics of surface hydrophilic with a thickness of 0.4mm, surface density 15g/m², surface static water contact angle is 65°；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture Measure as follows: fructose 5, peptone 0.5, yeast extract 0.5, citric acid 0.1, disodium hydrogen phosphate 0.2, potassium dihydrogen phosphate 0.1, surplus are Water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 production acetobacter in thaw at RT, be placed in 250mL fermentation Shaking table culture is carried out in culture solution, the revolving speed of shaking table culture is 150r/min, and temperature is 32 DEG C, and the time is for 24 hours；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:20, being stood at 32 DEG C Culture 9 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 8000r/ Min, centrifugation time 10min；

(6) prepare strain cultured solution: it is 5.0 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, continuously shakes at 2000rpm Dynamic 30s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 65 °, stationary culture 9 days at 32 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 80nm trunk bacteria cellulose fibre and average diameter is 20nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 20~60nm, with nanometer spider web structure bacteria cellulose film with a thickness of 9mm, density 8mg/cm³。

Embodiment 5

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through high steam It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the non-woven fabrics table being made of PAN material that average diameter is 25 μm for negatively charged hydrophilic radical Face, the non-woven fabrics of surface hydrophilic with a thickness of 0.8mm, surface density 20g/m², surface static water contact angle is 61 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: mannitol 3, peptone 0.1, yeast extract 0.2, citric acid 0.06, disodium hydrogen phosphate 0.08, potassium dihydrogen phosphate 0.05, Surplus is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 Pasteur's acetobacter in thaw at RT, be placed in 150mL hair Shaking table culture is carried out in ferment culture solution, the revolving speed of shaking table culture is 120r/min, and temperature is 28 DEG C, time 12h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:15, being stood at 29 DEG C Culture 7 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 5000r/ Min, centrifugation time 6min；

(6) prepare strain cultured solution: it is 4.9 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, continuously shakes at 1200rpm Dynamic 30s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 61 °, stationary culture 6 days at 31 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 65nm trunk bacteria cellulose fibre and average diameter is 16nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 60~90nm, with nanometer spider web structure bacteria cellulose film with a thickness of 7mm, density 7.2mg/cm³。

Embodiment 6

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through high steam It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the nonwoven being made of polyglycolide material that average diameter is 50 μm for negatively charged hydrophilic radical Cloth surface, the non-woven fabrics of surface hydrophilic with a thickness of 0.2mm, surface density 40g/m², surface static water contact angle is 50 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: mannitol 3.5, peptone 0.3, yeast extract 0.2, citric acid 0.06, disodium hydrogen phosphate 0.09, potassium dihydrogen phosphate 0.01, surplus is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 Agrobacterium in thaw at RT, be placed in 250mL fermentation training Shaking table culture is carried out in nutrient solution, the revolving speed of shaking table culture is 50r/min, and temperature is 32 DEG C, time 6h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:15, being stood at 31 DEG C Culture 7 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 6000r/ Min, centrifugation time 9min；

(6) prepare strain cultured solution: it is 4.7 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 1500 rpm continuous vibration Dynamic 18s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 50 °, stationary culture 3 days at 28 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 40nm trunk bacteria cellulose fibre and average diameter is 10nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 120~185nm, with nanometer spider web structure bacteria cellulose film with a thickness of 2mm, density 5.5mg/cm³。

Embodiment 7

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through ultraviolet irradiation It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the non-woven fabrics material table being made of PLA that average diameter is 20 μm for negatively charged hydrophilic radical Face, the non-woven fabrics of surface hydrophilic with a thickness of 0.5mm, surface density 35g/m², surface static water contact angle is 58 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: fructose 5, peptone 0.05, yeast extract 0.03, citric acid 0.05, disodium hydrogen phosphate 0.07, potassium dihydrogen phosphate 0.06, Surplus is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 rhizobium in thaw at RT, be placed in 180mL fermentation training Shaking table culture is carried out in nutrient solution, the revolving speed of shaking table culture is 110r/min, and temperature is 29 DEG C, time 18h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:12, being stood at 28 DEG C Culture 8 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 6000r/ Min, centrifugation time 8min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 800 rpm continuous vibration Dynamic 18s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 58 °, stationary culture 4 days at 30 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 55nm trunk bacteria cellulose fibre and average diameter is 12nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 150~200nm, with nanometer spider web structure bacteria cellulose film with a thickness of 4mm, density 6.5mg/cm³。

Embodiment 8

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, through Co 60-gamma It is laid in culture vessel after x ray irradiation x sterilization treatment, the hydrophilic non-woven fabrics in face is to be handled to make through surface hydrophilic by non-woven fabrics , surface hydrophilic processing, which refers to, is grafted on the nothing being made of PE material that average diameter is 40 μm for negatively charged hydrophilic radical Woven fabric surface, the non-woven fabrics of surface hydrophilic with a thickness of 1mm, surface density 15g/m², surface static water contact angle is 68 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture Measure as follows: sucrose 4, peptone 0.04, yeast extract 0.5, citric acid 0.1, disodium hydrogen phosphate 0.02, potassium dihydrogen phosphate 0.02 are remaining Amount is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 sarcine in thaw at RT, be placed in 150mL fermentation Shaking table culture is carried out in culture solution, the revolving speed of shaking table culture is 140r/min, and temperature is 31 DEG C, time 18h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:18, being stood at 30 DEG C Culture 6 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 6000r/ Min, centrifugation time 6min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 600 rpm continuous vibration Dynamic 18s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 68 °, stationary culture 3 days at 32 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 40nm trunk bacteria cellulose fibre and average diameter is 8nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 150~200nm, with nanometer spider web structure bacteria cellulose film with a thickness of 1.8mm, density 5.5mg/cm³。

Embodiment 9

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, ethane via epoxyethane goes out It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the non-woven fabrics table being made of PLGA material that average diameter is 45 μm for negatively charged hydrophilic radical Face, the non-woven fabrics of surface hydrophilic with a thickness of 0.1mm, surface density 40g/m², surface static water contact angle is 45 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture Measure as follows: sucrose 3, peptone 0.2, yeast extract 0.5, citric acid 0.01, disodium hydrogen phosphate 0.09, potassium dihydrogen phosphate 0.05 are remaining Amount is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 Pseudomonas cepacia in thaw at RT, be placed in 180mL Shaking table culture is carried out in fermentation culture, the revolving speed of shaking table culture is 100r/min, and temperature is 29 DEG C, time 20h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:8, being stood at 28 DEG C Culture 9 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 5000r/ Min, centrifugation time 4min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 800 rpm continuous vibration Dynamic 25s, obtaining strain cell density is 10⁹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 45 °, stationary culture 5 days at 28 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 45nm trunk bacteria cellulose fibre and average diameter is 10nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture is the bacteria cellulose film that 190~230nm has nanometer spider web structure with a thickness of 3.5mm, density 6mg/cm³。

Embodiment 10

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, through Co 60-gamma It is laid in culture vessel after x ray irradiation x sterilization treatment, the non-woven fabrics of surface hydrophilic is to be handled to make through surface hydrophilic by non-woven fabrics , surface hydrophilic processing refers to that it is 25 μm by chitosan undulation degree that negatively charged hydrophilic radical, which is grafted on average diameter, Texture at nonwoven surface, the non-woven fabrics of surface hydrophilic with a thickness of 0.9mm, surface density 25g/m², surface static water connects Feeler is 60 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: mannitol 2, peptone 0.5, yeast extract 0.08, citric acid 0.07, disodium hydrogen phosphate 0.06, potassium dihydrogen phosphate 0.03, Surplus is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 Pseudomonas cocovenenans in thaw at RT, be placed in 160mL Shaking table culture is carried out in fermentation culture, the revolving speed of shaking table culture is 80r/min, and temperature is 29 DEG C, time 9h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:12, being stood at 31 DEG C Culture 7 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 2000r/ Min, centrifugation time 6min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, continuously shakes at 1200rpm Dynamic 25s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 60 °, stationary culture 4 days at 32 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 55nm trunk bacteria cellulose fibre and average diameter is 13nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture is the bacteria cellulose film that 120~160nm has nanometer spider web structure with a thickness of 4mm, density 6.5mg/cm³。

Embodiment 11

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through high steam It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the nonwoven surface being made of PP material that average diameter is 50 μm for negatively charged hydrophilic radical, The non-woven fabrics of surface hydrophilic with a thickness of 0.7mm, surface density 40g/m², surface static water contact angle is 70 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture Measure as follows: fructose 3, peptone 0.09, yeast extract 0.3, citric acid 0.06, disodium hydrogen phosphate 0.15, potassium dihydrogen phosphate 0.06 are remaining Amount is water；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 campylobacter jejuni in thaw at RT, be placed in 230mL hair Shaking table culture is carried out in ferment culture solution, the revolving speed of shaking table culture is 120r/min, and temperature is 29 DEG C, time 21h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:18, being stood at 28 DEG C Culture 5 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 8000r/ Min, centrifugation time 3min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 500 rpm continuous vibration Dynamic 10s, obtaining strain cell density is 10¹¹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 70 °, stationary culture 7 days at 32 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 65nm trunk bacteria cellulose fibre and average diameter is 17nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 50~90nm, with nanometer spider web structure bacteria cellulose film with a thickness of 7.5mm, density 7.2mg/cm³。

Embodiment 12

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, ethane via epoxyethane goes out It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the non-woven fabrics table being made of PET material that average diameter is 40 μm for negatively charged hydrophilic radical Face, the non-woven fabrics of surface hydrophilic with a thickness of 0.8mm, surface density 30g/m², surface static water contact angle is 58 °；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 Pseudomonas cocovenenans and campylobacter jejuni (mass ratio 2: 3) it in thaw at RT, is placed in 230mL fermentation culture and carries out shaking table culture, the revolving speed of shaking table culture is 120r/min, temperature Degree is 29 DEG C, time 21h；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 500 rpm continuous vibration Dynamic 30s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 58 °, stationary culture 5 days at 32 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 60nm trunk bacteria cellulose fibre and average diameter is 15nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 90~130nm, with nanometer spider web structure bacteria cellulose film with a thickness of 5.5mm, density 7mg/cm³。

Embodiment 13

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, through Co 60-gamma It is laid in culture vessel after x ray irradiation x sterilization treatment, the non-woven fabrics of surface hydrophilic is to be handled to make through surface hydrophilic by non-woven fabrics , surface hydrophilic processing, which refers to negatively charged hydrophilic radical is grafted on average diameter to be 30 μm, to be made of PLA material Nonwoven surface, the non-woven fabrics of surface hydrophilic with a thickness of 1mm, surface density 20g/m², surface static water contact angle is 50 °；

(3) activate: by the isoelectric point of refrigeration be 3.5~4.0 gluconacetobacter, produce acetobacter and acetify bacillus (quality Than being 2:1:2) in thaw at RT, it is placed in 230mL fermentation culture and carries out shaking table culture, the revolving speed of shaking table culture is 120r/min, temperature are 29 DEG C, time 21h；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 1500 rpm continuous vibration Dynamic 18s, obtaining strain cell density is 10¹⁰The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the non-woven fabrics that angle is 50 °, stationary culture 6 days at 28 DEG C obtain having the bacteria cellulose film of nanometer spider web structure compound Material.

The bacteria cellulose film composite material with nanometer spider web structure prepared is double-layer structure, respectively has and receives The rice bacteria cellulose film of spider web structure and the non-woven fabrics of surface hydrophilic, the bacteria cellulose film with nanometer spider web structure are main The mycobacterium cellulose fibre that by average diameter be 61nm trunk bacteria cellulose fibre and average diameter is 16nm is constituted, main Dry bacteria cellulose fibre is the skeleton to form three-dimensional mesh structure and constitute bacteria cellulose film that is cross-linked with each other between itself Bacteria cellulose fibre；Mycobacterium cellulose fibre is that bacteria cellulose into the net is fine between trunk bacteria cellulose fibre Dimension.Trunk bacteria cellulose fibre and mycobacterium cellulose fibre pass through β -1,4- glucosides key connection.The mesh of nanometer spider web Aperture be 120~160nm, with nanometer spider web structure bacteria cellulose film with a thickness of 6mm, density 6.8mg/cm³。

Embodiment 14

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through ultraviolet irradiation It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the non-woven fabrics table being made of viscose glue material that average diameter is 20 μm for negatively charged hydrophilic radical Face, the non-woven fabrics of surface hydrophilic with a thickness of 0.5mm, surface density 15g/m², surface static water contact angle is 70 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: glucose 4, peptone 0.25, yeast extract 0.1, citric acid 0.06, disodium hydrogen phosphate 0.12, potassium dihydrogen phosphate 0.01, Surplus is water；

(3) it activates: being 3.5~4.0 to acetify bacillus and sarcine (mass ratio 2:1) in room by the isoelectric point of refrigeration Temperature is thawed, and is placed in 180mL fermentation culture and is carried out shaking table culture, and the revolving speed of shaking table culture is 90r/min, temperature 30 DEG C, time 16h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:3, being stood at 28 DEG C Culture 9 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 7000r/ Min, centrifugation time 8min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, continuously shakes at 2000rpm Dynamic 25s, obtaining strain cell density is 10⁹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the one of surface for the non-woven fabrics that angle is 70 °, stationary culture 6 days at 28 DEG C.

(8) another surface of non-woven fabrics continues to cultivate: culture bacterium solution obtained in step (7) being added dropwise and is connect in surface static water On another surface for the non-woven fabrics that feeler is 70 °, stationary culture 6 days at 28 DEG C.

The bacteria cellulose film composite material with nanometer spider web structure prepared is three-decker, from top to bottom successively For the non-woven fabrics of bacteria cellulose film, surface hydrophilic with nanometer spider web structure and the bacterial fibers with nanometer spider web structure Plain film, the bacteria cellulose film with nanometer spider web structure are mainly 40nm trunk bacteria cellulose fibre peace by average diameter The mycobacterium cellulose fibre that equal diameter is 20nm is constituted, and trunk bacteria cellulose fibre is to be cross-linked with each other to be formed between itself Three-dimensional mesh structure and constitute bacteria cellulose film skeleton bacteria cellulose fibre；Mycobacterium cellulose fibre is in master Bacteria cellulose fibre into the net between dry bacteria cellulose fibre.Trunk bacteria cellulose fibre and mycobacterium cellulose fiber Dimension passes through β -1,4- glucosides key connection.The mesh aperture of nanometer spider web is 250~300nm, the bacterium with nanometer spider web structure Cellulose membrane with a thickness of 9mm, density 6mg/cm³。

Embodiment 15

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is gone out through ultraviolet irradiation It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the nonwoven being made of polyglycolide material that average diameter is 25 μm for negatively charged hydrophilic radical Cloth surface, the non-woven fabrics of surface hydrophilic with a thickness of 0.4mm, surface density 15g/m², surface static water contact angle is 69 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture Measure as follows: sucrose 2, peptone 0.45, yeast extract 0.05, citric acid 0.1, disodium hydrogen phosphate 0.18, potassium dihydrogen phosphate 0.03 are remaining Amount is water；

(3) it activates: being 3.5~4.0 grape saccharobacillus and Agrobacterium (mass ratio 1:1) in room by the isoelectric point of refrigeration Temperature is thawed, and is placed in 150mL fermentation culture and is carried out shaking table culture, and the revolving speed of shaking table culture is 90r/min, temperature 32 DEG C, time 6h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:7, being stood at 28 DEG C Culture 9 days；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, at 1500 rpm continuous vibration Dynamic 15s, obtaining strain cell density is 10¹²The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the one of surface for the non-woven fabrics that angle is 69 °, stationary culture 4 days at 32 DEG C.

(8) another surface of non-woven fabrics continues to cultivate: culture bacterium solution obtained in step (7) being added dropwise and is connect in surface static water On another surface for the non-woven fabrics that feeler is 69 °, stationary culture 4 days at 32 DEG C.

The bacteria cellulose film composite material with nanometer spider web structure prepared is three-decker, from top to bottom successively For the non-woven fabrics of bacteria cellulose film, surface hydrophilic with nanometer spider web structure and the bacterial fibers with nanometer spider web structure Plain film, the bacteria cellulose film with nanometer spider web structure are mainly 40nm trunk bacteria cellulose fibre peace by average diameter The mycobacterium cellulose fibre that equal diameter is 5nm is constituted, and trunk bacteria cellulose fibre is to be cross-linked with each other to be formed between itself Three-dimensional mesh structure and constitute bacteria cellulose film skeleton bacteria cellulose fibre；Mycobacterium cellulose fibre is in master Bacteria cellulose fibre into the net between dry bacteria cellulose fibre.Trunk bacteria cellulose fibre and mycobacterium cellulose fiber Dimension passes through β -1,4- glucosides key connection.The mesh aperture of nanometer spider web is 20~70nm, and the bacterium with nanometer spider web structure is fine Tie up plain film with a thickness of 5mm, density 5mg/cm³。

Embodiment 16

(1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, ethane via epoxyethane goes out It is laid in culture vessel after bacterium processing, the non-woven fabrics of surface hydrophilic is to be handled to be made through surface hydrophilic by non-woven fabrics, surface parent Water process, which refers to, is grafted on the non-woven fabrics table being made of PET material that average diameter is 35 μm for negatively charged hydrophilic radical Face, the non-woven fabrics of surface hydrophilic with a thickness of 0.6mm, surface density 40g/m², surface static water contact angle is 66 °；

(2) prepare fermentation culture: by mass percentage, unit wt% each component and its contains in fermentation culture It measures as follows: mannitol 5, peptone 0.45, yeast extract 0.05, citric acid 0.04, disodium hydrogen phosphate 0.1, potassium dihydrogen phosphate 0.03, Surplus is water；

(3) it activates: gluconacetobacter, rhizobium and the Pseudomonas cocovenenans (matter for being 3.5~4.0 by the isoelectric point of refrigeration Amount is than being 2:3:3) in thaw at RT, it is placed in 100mL fermentation culture and carries out shaking table culture, the revolving speed of shaking table culture is 50r/min, temperature are 28 DEG C, time 16h；

(4) it expands: after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:2, being stood at 29 DEG C Culture 8 days；

(5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution, centrifugal rotational speed 3000r/ Min, centrifugation time 10min；

(7) it cultivates: strain cultured solution being added in concentration bacterium solution, and uses eddy blending machine, continuously shakes at 900rpm Dynamic 10s, obtaining strain cell density is 10⁹The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise and is contacted in surface static water On the one of surface of non-woven fabrics that angle is 66 °, stationary culture 5 days at 28 DEG C obtain the bacterium with nanometer spider web structure Cellulose membrane composite material.

(8) another surface of non-woven fabrics continues to cultivate: culture bacterium solution obtained in step (7) being added dropwise and is connect in surface static water On another surface for the non-woven fabrics that feeler is 66 °, stationary culture 5 days at 28 DEG C.

The bacteria cellulose film composite material with nanometer spider web structure prepared is three-decker, from top to bottom successively For the non-woven fabrics of bacteria cellulose film, surface hydrophilic with nanometer spider web structure and the bacterial fibers with nanometer spider web structure Plain film, the bacteria cellulose film with nanometer spider web structure are mainly 80nm trunk bacteria cellulose fibre peace by average diameter The mycobacterium cellulose fibre that equal diameter is 10nm is constituted, and trunk bacteria cellulose fibre is to be cross-linked with each other to be formed between itself Three-dimensional mesh structure and constitute bacteria cellulose film skeleton bacteria cellulose fibre；Mycobacterium cellulose fibre is in master Bacteria cellulose fibre into the net between dry bacteria cellulose fibre.Trunk bacteria cellulose fibre and mycobacterium cellulose fiber Dimension passes through β -1,4- glucosides key connection.The mesh aperture of nanometer spider web is 240~300nm, the bacterium with nanometer spider web structure Cellulose membrane with a thickness of 9mm, density 4mg/cm³。

Claims

1. the bacteria cellulose film composite material with nanometer spider web structure, it is characterized in that: with two layers or more of structure, In, adjacent two layers are respectively to have a nanometer non-woven fabrics for the bacteria cellulose film of spider web structure and surface hydrophilic；

The bacteria cellulose film with nanometer spider web structure is mainly by trunk bacteria cellulose fibre and mycobacterium fiber Cellulose fiber is constituted；

The trunk bacteria cellulose fibre is to be cross-linked with each other to form three-dimensional mesh structure and constitute bacteria cellulose between itself The bacteria cellulose fibre of the skeleton of film；

2. the bacteria cellulose film composite material according to claim 1 with nanometer spider web structure, which is characterized in that institute It states trunk bacteria cellulose fibre and mycobacterium cellulose fibre and passes through β -1,4- glucosides key connection.

3. the bacteria cellulose film composite material according to claim 1 with nanometer spider web structure, which is characterized in that institute The mesh aperture for stating nanometer spider web is 20~300nm；The bacteria cellulose film with nanometer spider web structure with a thickness of 0.1 ~9mm, density are 5~8mg/cm³；The average diameter of the trunk bacteria cellulose fibre is 30~80nm；The branch is thin The average diameter of fungin fiber is 5~20nm.

4. the bacteria cellulose film composite material according to claim 1 with nanometer spider web structure, which is characterized in that institute State the non-woven fabrics of surface hydrophilic with a thickness of 0.1~1mm, surface density is 15~40g/m², by non-woven fabrics through surface hydrophilic Reason is made；

5. the bacteria cellulose film composite material according to claim 4 with nanometer spider web structure, which is characterized in that institute It is that 20~50 μm of fiber is constituted that state non-woven fabrics, which be by average diameter, material be polypropylene, polyethylene terephthalate, Polyamide, viscose glue, polyacrylonitrile, polyglycolide, polylactic acid, polyethylene, polylactide glycolate copolymer or chitosan；

6. preparing the bacteria cellulose film composite material as claimed in any one of claims 1 to 5 with nanometer spider web structure Method, it is characterized in that: the bacterial strain that isoelectric point is 3.5~4.0 is dispersed in the strain cultured solution that pH value is 4.5~5.0 Obtaining strain cell density is 10⁹~10¹²The culture bacterium solution of a/mL, then culture bacterium solution is added dropwise in surface static water contact angle The bacteria cellulose film composite material with nanometer spider web structure is made in stationary culture on the non-woven fabrics of≤70 ° of surface hydrophilic.

7. according to the method described in claim 6, it is characterized in that, specifically includes the following steps:

1) sterilization treatment is carried out to non-woven fabrics: the non-woven fabrics of surface hydrophilic is cut into required size, is tiled after sterilized processing In in culture vessel；

2) prepare fermentation culture: by mass percentage, unit wt%, each component and its content be such as in fermentation culture Under:

3) activate: by the isoelectric point of refrigeration be 3.5~4.0 bacterial strain in thaw at RT, be placed in 100~250mL fermented and cultured Carrying out shaking table culture in liquid, the revolving speed of shaking table culture is 50~150r/min, and temperature is 28~32 DEG C, the time is 6~for 24 hours；

4) it expands: quiet at 28~32 DEG C after the bacterium solution that activation finishes is mixed with fermentation culture with the volume ratio of 1:2~20 Set culture 5~9 days；

5) it is centrifuged: by the bacterium solution centrifugation removal supernatant after amplification, obtaining concentration bacterium solution；

6) prepare strain cultured solution: it is 4.5~5.0 that sodium hydroxide is added in fermentation culture and adjusts pH, obtains strain culturing Liquid；

7) it cultivates: strain cultured solution is added in concentration bacterium solution, and evenly dispersed, obtaining strain cell density is 10⁹~10¹² The culture bacterium solution of a/mL；Then culture bacterium solution is added dropwise on the non-woven fabrics of surface static water contact angle≤70 °, 28~32 Stationary culture 3~9 days at DEG C obtain the bacteria cellulose film composite material with nanometer spider web structure.

8. according to the method described in claim 6, it is characterized in that, the bacterial strain that the isoelectric point is 3.5~4.0 is selected from glucose vinegar Bacillus produces acetobacter, acetifies the false list of bacillus, Pasteur's acetobacter, grape saccharobacillus, Agrobacterium, rhizobium, sarcine, onion One or more of born of the same parents bacterium, Pseudomonas cocovenenans or campylobacter jejuni.

9. the method according to the description of claim 7 is characterized in that the sterilization treatment refers to high pressure steam sterilization, ultraviolet spoke According to sterilizing, ethylene oxide sterilizing or Co 60-radiated by gamma-ray sterilizing；The centrifugal rotational speed of the centrifugation is 2000~8000r/ Min, centrifugation time are 3~10min；It is described it is evenly dispersed refer to using eddy blending machine, continuously shake at 500~2000rpm Dynamic 10~30s.