CN106755179B - A kind of culture medium suitable for bacteria cellulose fermentation - Google Patents
A kind of culture medium suitable for bacteria cellulose fermentation Download PDFInfo
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- CN106755179B CN106755179B CN201611113691.1A CN201611113691A CN106755179B CN 106755179 B CN106755179 B CN 106755179B CN 201611113691 A CN201611113691 A CN 201611113691A CN 106755179 B CN106755179 B CN 106755179B
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- C12P19/00—Preparation of compounds containing saccharide radicals
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Abstract
The invention belongs to fermentation technical fields, and in particular to a kind of culture medium suitable for bacteria cellulose fermentation.The culture medium is using corn flour, analysis for soybean powder, bean cake powder as major ingredient, not only reduce cost, and raw material it is cheap and easy to get, it is full of nutrition comprehensively, processing technology it is simple, more conventional 5-7 times of fermentation medium for bacterial cellulose cost reduction, and the bacteria cellulose fermentation yield that 24g/L or more can be completed was cultivated by 136-144 hours, improve the production efficiency of cellulose.
Description
Technical field
The invention belongs to fermentation technical fields, and in particular to a kind of culture medium suitable for bacteria cellulose fermentation.
Technical background
Bacteria cellulose is that certain monoid microorganism (bacterium) economic cooperation is referred to as at the cellulose of secretion, mostly with wood in industrial production
Acetobacter makees the strain of bacteria cellulose fermentation.As a kind of novel biomaterial, bacteria cellulose with its special performance by
Extensive concern.Bacteria cellulose have high-purity, ultra-fine property, permeable breathable, good biocompatibility, degradability and
Biosynthesis Modulatory character, in medical instruments field, food additives field, field of papermaking, field of textiles and field of environment protection
Have broad application prospects.
Microbial fermentation is complicated, system a biological process, is during which influenced by many factors, such as culture medium
Component, proportioning, cultivation temperature, training method.Wherein, the composition and ratio of culture medium are one of key factors, both at home and abroad into
Numerous studies are gone.
Domestic bacteria cellulose production is initially using coconut water as primary raw material, and such as application No. is 201110223884.3 hairs
A kind of fermentation medium for bacterial cellulose is disclosed in bright application, is contained in the culture medium and is accounted for culture medium total weight 30-90%'s
The coconut dregs of rice add the slurries obtained after water mixing.This method is not easy to wide popularization and application by territory restriction.Scientific research and production people
Member successively develops juice, plant extract and all kinds of agricultural and sideline product leftover bits and pieces etc. as bacteria cellulose fermentation major ingredient.
Current media has the following disadvantages：1) feed preparation process is complicated, expends larger man power and material.2) raw material by
Territory restriction is unfavorable for promoting.3) bacteria cellulose yield is relatively low, increases the unit cost of production, high production cost limit
The popularization and application of bacteria cellulose are made, current bacteria cellulose is mainly limited to the higher product scope of added value.
China is Chan Liang big countries, possesses abundant cultivated land resource, and corn is the third-largest cereal crops in China, resourceful,
It is quality-high and inexpensive.Corn contains abundant starch, protein and trace element, is ideal culture medium raw material, such as China
A kind of preparation methods of bacteria cellulose of patent of invention ZL201110110794.3 disclose one kind and adding corn in the medium
For core enzymolysis liquid come the method for preparing bacteria cellulose, this method cellulase, hemicellulase and zytase digest corn
Corncob enzymolysis liquid is made in core, and adds it in the culture medium for preparing bacteria cellulose, can make to be rich in corncob
Xylan is digested into the xylo-oligosaccharide with good healthcare function, and is wrapped or is adsorbed in bacteria cellulose, and bacterium is improved
The healthcare function of cellulose.But not yet find the technology using corn powder production bacteria cellulose.
Soybean is one of important cereal crops in China, and containing abundant protein, good carbon can be provided for microorganism
The nutrient raw materials such as source, nitrogen source.By-product -- the dregs of beans of beans processing because containing abundant carbohydrate and protein and
Trace element is also ideal microbial fermentation raw material.
It is applied to the report of bacteria cellulose fermentation currently without corn powder, analysis for soybean powder and dregs of beans.
Invention content
Present invention aims at：A kind of applicable culture medium significantly improving bacteria cellulose output is provided, used in culture medium
Raw material is cheap and easy to get, it is full of nutrition comprehensively, processing technology it is simple.While reducing cost of material, bacterium fibre is significantly improved
The plain yield of dimension, is greatly lowered the production cost of bacteria cellulose, effectively facilitates the popularization and application of bacteria cellulose.The present invention uses
The major ingredient of corn flour, analysis for soybean powder, bean cake powder as bacteria cellulose fermented and cultured, not only reduces cost, and nutrition is comprehensive,
It is easily detached with product, is conducive to the volume increase and collection of bacteria cellulose.
The culture medium is specific as follows：
(1) seed culture medium：1000 parts by weight meters, 5-15 parts of corn flour, 5-12 parts of analysis for soybean powder, 5-10 parts of white granulated sugar,
1-3 parts of yeast extract, 1-3 parts of tryptone, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, surplus are water, and pH to 6.8,121 DEG C go out
Bacterium 20 minutes；
(2) fermentation medium：1000 parts by weight meters, 8-20 parts of corn flour, 5-20 parts of bean cake powder, 5-15 parts of white granulated sugar,
2 parts of potassium dihydrogen phosphate, 2 parts of magnesium sulfate, 1 part of calcium chloride, surplus are water, and pH to 6.8,121 DEG C sterilize 20 minutes；
The corn flour is crushed to 60 mesh and following by whole kernel corn；
The analysis for soybean powder is crushed to 60 mesh and following by soybean grain；
The bean cake powder is a leaching dregs of beans, is crushed to 60 mesh and following；
The white granulated sugar is food grade certified products；
The yeast extract, tryptone, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride are commercially available conventional reagent；
The present invention also provides a kind of methods preparing bacteria cellulose using above-mentioned culture medium fermentation：
(1) strain is accessed after routinely expanding numerous tube in above-mentioned seed culture fluid；
The strain can be xylose gluconic acid acetobacter (Gluconacetobacter xylinus), acetobacter aceti
(Acetobacter aceti) Pasteur's acetobacter (Acetobacterpasteutinus), Hansen acetobacter
(Acetobacterhansenii) or tunica albuginea acetobacter (Acetobacter acetosus)；
(2) seed culture condition is：25 DEG C in constant-temperature table, 120-150rpm cultivate 40-48 hour obtain seed liquor；
(3) bacteria cellulose fermenting and producing is controlled using three-stage：
First stage：Seed liquor is seeded in fermentation medium by the inoculum concentration of 5-10%, 26-28 DEG C, 120-
180rpm is cultivated 24 hours；
Second stage：Cultivation temperature is adjusted to 28-30 DEG C, 150-240rpm is cultivated 48 hours；
Phase III：28-30 DEG C, 80-160rpm continue culture 48 hours, obtain bacteria cellulose dynamic fermentation particle；
(4) the bacteria cellulose dynamic fermentation particle for obtaining fermented and cultured is rinsed through deionized water, 1%NaOH solution soaks
Boil, deionized water is rinsed to neutrality, after freeze-drying target product.
Advantageous effect：
(1) culture medium provided by the invention it is raw materials used it is cheap and easy to get, full of nutrition comprehensively, processing technology it is simple；It is dropping
While low cost of material, bacteria cellulose output is significantly improved to 24g/L or more, effectively increases bacteria cellulose
The production cost of bacteria cellulose is greatly lowered in yield, effectively facilitates the popularization and application of bacteria cellulose；
(2) present invention use corn flour, analysis for soybean powder, bean cake powder as bacteria cellulose fermented and cultured major ingredient, it is more conventional carefully
5-7 times of fungin fermentation medium cost reduction, and nutrition is comprehensive, is easily detached with product, is conducive to bacteria cellulose
Cleaning and collection；
(3) culture medium provided by the present invention is not only at low cost, yield is high, but also improves the production efficiency of cellulose,
The bacteria cellulose fermentation yield that 24g/L or more can be completed was cultivated by 136-144 hours.
(4) bacteria cellulose fermentation process of the invention uses the fermentation process of three-stage temperature-speed-changing, according to bacterium for the first time
The inoculum concentration and metabolic alterations of kind adjust temperature and rotating speed, easy to operate, effectively increase the yield of bacteria cellulose.
Description of the drawings：
Fig. 1：Bacteria cellulose flow sheet
Specific implementation mode：
Strain used is xylose gluconic acid acetobacter (Gluconacetobacter xylinus) in following embodiment
CCTCC AB2040512；
Every sampling observation in 24 hours, simultaneously analytical unit volume biomass was primary during embodiment fermentation tank dynamic cultivation,
Using being observed under Gram's stain smear microscopy.
A kind of method of the culture medium and fermented-producing bacteria cellulose suitable for bacteria cellulose fermentation of embodiment 1
Seed culture medium：In terms of 1000 parts, 15 parts of corn flour, 5 parts of analysis for soybean powder, 5 parts of white granulated sugar, 1 part of tryptone, yeast
2 parts of medicinal extract, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes；
Fermentation medium：In terms of 1000 parts, 20 parts of corn flour, 6 parts of bean cake powder, 5 parts of white granulated sugar, 2 parts of potassium dihydrogen phosphate, sulphur
2 parts of sour magnesium, 1 part of calcium chloride, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes；
The corn flour is crushed to 60 mesh and following by whole kernel corn；The analysis for soybean powder by soybean grain be crushed to 60 mesh and with
Under；The bean cake powder is a leaching dregs of beans, is crushed to 60 mesh and following；The white granulated sugar is food grade certified products；The yeast leaching
Cream, tryptone, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride are commercially available conventional reagent.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 150rpm in constant-temperature table
It cultivates 48 hours, it is spare.
The bacteria cellulose dynamic fermentation first stage：Seed is seeded to by 10% inoculum concentration in fermentation tank, 180rpm,
28 DEG C are cultivated 24 hours；Second stage：By condition of culture be adjusted to 30 DEG C, 210rpm cultivate 48 hours；Phase III：By rotating speed
It is filtered to remove bacterium solution after being adjusted to 140,28 DEG C of cultures 48 hours and obtains bacteria cellulose granular materials.
Product processing：Gained granular materials is cleaned with deionized water, 1% hydrogen-oxygen of mass concentration is used after cleaning remaining medium
Change 90 DEG C of sodium solution to handle 15 minutes, deionized water rinses to neutral and obtains BC (bacteria cellulose) moist wood, and resulting materials are freezed
Dry BC dry powder, weighs and records.
A kind of method of the culture medium and fermented-producing bacteria cellulose suitable for bacteria cellulose fermentation of embodiment 2
Seed culture medium：In terms of 1000 parts, 10 parts of corn flour, 12 parts of analysis for soybean powder, 6 parts of white granulated sugar, 1 part of yeast extract, pancreas
1 part of peptone, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes.
Fermentation medium：In terms of 1000 parts, 14 parts of corn flour, 8 parts of bean cake powder, 10 parts of white granulated sugar, 2 parts of potassium dihydrogen phosphate,
2 parts of magnesium sulfate, 1 part of calcium chloride, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 150rpm in constant-temperature table
It cultivates 48 hours, it is spare.
Bacteria cellulose dynamic fermentation：
Seed is seeded to by 6% inoculum concentration in fermentation tank, 140rpm, 27 DEG C cultivate 24 hours；Second stage：It will training
Foster condition be adjusted to 29 DEG C, adjustment of rotational speed be 240rpm cultivate 48 hours；Phase III：Temperature is adjusted to 30 DEG C, rotating speed tune
Whole is to be filtered to remove bacterium solution after 80rpm is cultivated 48 hours to obtain bacteria cellulose granular materials.
Other processing modes such as embodiment 1.
A kind of method of the culture medium and fermented-producing bacteria cellulose suitable for bacteria cellulose fermentation of embodiment 3
Seed culture medium is prepared：In terms of 1000 parts, 5 parts of corn flour, 10 parts of analysis for soybean powder, 7 parts of white granulated sugar, 2 parts of tryptone,
2 parts of yeast extract, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes.
Fermentation medium is prepared：In terms of 1000 parts, 10 parts of corn flour, 14 parts of bean cake powder, 12 parts of white granulated sugar, potassium dihydrogen phosphate
2 parts, 2 parts of magnesium sulfate, 1 part of calcium chloride, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 130rpm in constant-temperature table
It cultivates 40 hours, it is spare.
Bacteria cellulose dynamic fermentation the first valence section：Seed is seeded to by 5% inoculum concentration in fermentation tank, setting speed
For 120rpm, cultivation temperature is set as 26 DEG C and cultivates 24 hours；Second stage：It is 180rpm, culture by fermentation tank adjustment of rotational speed
Temperature is adjusted to 28 DEG C and cultivates 48 hours；Phase III：By adjustment of rotational speed 160rpm, 29 DEG C of cultures are filtered to remove bacterium after 48 hours
Liquid obtains bacteria cellulose granular materials.
Other processing modes such as embodiment 1.
A kind of method of the culture medium and fermented-producing bacteria cellulose suitable for bacteria cellulose fermentation of embodiment 4
Seed culture medium：In terms of 1000 parts, 11 parts of corn flour, 7 parts of analysis for soybean powder, 10 parts of white granulated sugar, 3 parts of yeast extract, pancreas
3 parts of peptone, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes.
Fermentation medium：In terms of 1000 parts, 8 parts of corn flour, 20 parts of bean cake powder, 15 parts of white granulated sugar, 2 parts of potassium dihydrogen phosphate,
2 parts of magnesium sulfate, 1 part of calcium chloride, surplus is water, adjusts pH to 6.8, and 121 DEG C sterilize 20 minutes.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 120rpm in constant-temperature table
It cultivates 48 hours, it is spare.
The bacteria cellulose dynamic fermentation first stage：Seed is seeded to by 8% inoculum concentration in fermentation tank, 160rpm,
28 DEG C are cultivated 24 hours；Second stage：By condition of culture be adjusted to 30 DEG C, 150rpm cultivate 48 hours；Phase III：29℃,
110rpm is filtered to remove bacterium solution after cultivating 48 hours and obtains bacteria cellulose granular materials.
Other processing modes such as embodiment 1.
5 check experiment 1 of embodiment
Seed culture medium：Glucose 20g/L, peptone 5g/L, potassium dihydrogen phosphate 4.3g, magnesium sulfate 2.5g, sodium chloride 3g,
Surplus is water, adjusts pH to 6.8；
Fermentation medium：Spontaneous fermentation coconut water 1000ml, sucrose 40g, yeast extract 10g, potassium dihydrogen phosphate 4.3g, sulphur
Sour magnesium 2.5g, sodium chloride 3g, surplus are water, adjust pH to 6.8.
Other processing modes such as embodiment 1.
6 check experiment 2 of embodiment
Seed culture medium and fermentation medium are prepared with embodiment 1, and seed culture uses 7% with embodiment 1, fermented and cultured
Inoculum concentration be seeded in fermentation tank, 30 DEG C, 120rpm is filtered to remove bacterium solution and obtains bacteria cellulose granular material after cultivating 120 hours
Material.
Table 1：Each test group fermentation medium cost
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 | Embodiment 6 |
| Culture medium cost (member/L) | 0.262 | 0.275 | 0.337 | 0.317 | 1.421 | 0.262 |
| Yield (dry weight g/L) | 26.73 | 24.88 | 28.69 | 25.66 | 1.95 | 12.31 |
Claims (3)
1. a kind of method preparing bacteria cellulose, which is characterized in that be as follows：
(1) seed culture：Strain is inoculated in seed culture medium, and 25 DEG C in constant-temperature table, 120-150rpm cultivates 40-48 hours；
(2) three-stage fermented and cultured：
First stage：Seed liquor is seeded in fermentation medium by the inoculum concentration of 5-10%, 26-28 DEG C, 120-180rpm trainings
It supports 24 hours；
Second stage：Cultivation temperature is adjusted to 28-30 DEG C, 150-240rpm is cultivated 48 hours；
Phase III：28-30 DEG C, 80-160rpm continue culture 48 hours, obtain bacteria cellulose dynamic fermentation particle；
(3) by fermented and cultured obtain bacteria cellulose dynamic fermentation particle through deionized water flushing, 1%NaOH solution digestion,
Deionized water is rinsed to neutrality, and bacterial cellulose product is obtained after freeze-drying；
The culture medium is specific as follows：
(1) seed culture medium：1000 parts by weight meters, 5-15 parts of corn flour, 5-12 parts of analysis for soybean powder, 5-10 parts of white granulated sugar, yeast
1-3 parts of medicinal extract, 1-3 parts of tryptone, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, surplus are water, pH to 6.8,121 DEG C of sterilizings 20
Minute；
(2) fermentation medium：1000 parts by weight meters, 8-20 parts of corn flour, 5-20 parts of bean cake powder, 5-15 parts of white granulated sugar, phosphoric acid
2 parts of potassium dihydrogen, 2 parts of magnesium sulfate, 1 part of calcium chloride, surplus are water, and pH to 6.8,121 DEG C sterilize 20 minutes.
2. the method as described in claim 1 for preparing bacteria cellulose, which is characterized in that the strain is selected from xylose gluconic acid
Acetobacter (Gluconacetobacter xylinus), acetobacter aceti (Acetobacter aceti) Pasteur's acetobacter
(Acetobacterpasteutinus), Hansen acetobacter (Acetobacterhansenii) or tunica albuginea acetobacter
(Acetobacter acetosus)。
3. application of the culture medium described in claim 1 in fermented-producing bacteria cellulose.
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| CN109984291A (en) * | 2018-01-03 | 2019-07-09 | 钟宇光 | A kind of preparation method of five cereals various grains convenient congee |
| CN109984193A (en) * | 2018-01-03 | 2019-07-09 | 钟宇光 | A kind of preparation method of fermented type coarse cereals Yoghourt |
| CN112877384A (en) * | 2021-03-26 | 2021-06-01 | 哈尔滨理工大学 | Preparation method of bacterial cellulose, bacterial cellulose-chitosan composite gel skin-care mask and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101319242A (en) * | 2008-06-25 | 2008-12-10 | 许春元 | Method for preparing bacteria cellulose with association of activity and inertia |
| CN102994430A (en) * | 2012-12-08 | 2013-03-27 | 福建卫生职业技术学院 | Bacterial cellulose production strain and application thereof |
| CN104031956A (en) * | 2014-06-05 | 2014-09-10 | 陕西科技大学 | Bacterial cellulose fermentation medium made from apple pomace and method for producing bacterial cellulose by utilizing medium |
| CN105132331A (en) * | 2015-09-18 | 2015-12-09 | 南京荣之盛生物科技有限公司 | Komagataeibacter nataicola and application thereof |
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- 2016-12-07 CN CN201611113691.1A patent/CN106755179B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101319242A (en) * | 2008-06-25 | 2008-12-10 | 许春元 | Method for preparing bacteria cellulose with association of activity and inertia |
| CN102994430A (en) * | 2012-12-08 | 2013-03-27 | 福建卫生职业技术学院 | Bacterial cellulose production strain and application thereof |
| CN104031956A (en) * | 2014-06-05 | 2014-09-10 | 陕西科技大学 | Bacterial cellulose fermentation medium made from apple pomace and method for producing bacterial cellulose by utilizing medium |
| CN105132331A (en) * | 2015-09-18 | 2015-12-09 | 南京荣之盛生物科技有限公司 | Komagataeibacter nataicola and application thereof |
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