CN105779345A - Method for promoting growth of zymomonas mobilis by using philodina culture solution - Google Patents

Method for promoting growth of zymomonas mobilis by using philodina culture solution Download PDF

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Publication number
CN105779345A
CN105779345A CN201610184834.1A CN201610184834A CN105779345A CN 105779345 A CN105779345 A CN 105779345A CN 201610184834 A CN201610184834 A CN 201610184834A CN 105779345 A CN105779345 A CN 105779345A
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flocculability
spinning roller
culture fluid
roller worm
antibacterial
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CN105779345B (en
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丁国际
林玮
胡梦楠
崔海涛
涂玉娟
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University of Shanghai for Science and Technology
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/70—Undefined extracts
    • C12N2500/80—Undefined extracts from animals
    • C12N2500/82—Undefined extracts from animals from invertebrates

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  • Life Sciences & Earth Sciences (AREA)
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  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)

Abstract

The invention discloses a method for promoting the growth of zymomonas mobilis by using a philodina culture solution. The method comprises the following steps of firstly, culturing philodina by using a culture solution with flour as bait; when the density of the philodina reaches 100 to 500ind/mL, filtering the philodina culture solution by using a 300-mesh nylon net, and removing philodina from the philodina culture solution; secondly, filtering and separating obtained filtrate by using an ultrafiltration membrane, and further removing fine particulate matters such as flour and the like in the filtrate, thus obtaining filtrate, namely the philodina culture solution. The consumption of the philodina culture solution, disclosed by the invention, serving as a zymomonas mobilis growth promoter is 0.05 to 1 percent. The method for promoting the growth of the zymomonas mobilis by using the philodina culture solution, disclosed by the invention, has the characteristics of ecology, convenience in use and remarkable effects.

Description

Utilize the method that spinning roller worm culture fluid promotes flocculability bacterial growth
Technical field
The present invention relates to a kind of dirty water and wastewater treatment bioremediation, particularly relate to a kind of method improving activated sludge flocculability, be applied to microorganism culturing field and sewage and biological wastewater treatment field.
Background technology
Activated sludge process is most widely used method in dirty water and wastewater treatment.The flocculability of activated sludge is one of key factor affecting the operation stability of active sludge processing system, treatment effeciency and effluent quality.The flocculability of activated sludge is more good, and the performance of active sludge processing system is more good.The flocculability of activated sludge depends on quantity and the activity of flocculability antibacterial, and the quantity increasing flocculability antibacterial or the activity improving flocculability antibacterial are all conducive to the raising of activated sludge flocculability.
The method improving activated sludge flocculability is segmented into two big classes: chemical method and biological methods.Chemical method is by adding inorganic flocculating agent or organic flocculant, increases the volume of activated sludge or increases the density of activated sludge, making the flocculability of activated sludge be improved.The side effect of this method is to make salinity in water increase, be easily formed secondary pollution, cause degradation problem under flocculability bacterial activity.Biological methods is by promoting that the flocculation activity of the growth of flocculability antibacterial or raising flocculability antibacterial improves the flocculability of activated sludge.The Chinese invention patent that patent documentation publication number is CN102816797 discloses the method by using Cys to strengthen light zymogenous bacteria excrement rhodopseudomonas flocculating property, but the method also exists the problems such as use specific and Cys the price of object is higher.
Summary of the invention
In order to solve prior art problem, it is an object of the invention to the deficiency overcoming prior art to exist, a kind of method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth is provided, utilize spinning roller worm culture fluid to improve the growth of flocculability antibacterial, simple to operate, ecological, easy to use, it is possible to increase the density of flocculability antibacterial, improve the flocculability of antibacterial, be conducive to extensive use, there are good economic benefits.
Create purpose for reaching foregoing invention, adopt following technical proposals:
A kind of method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth, by adding spinning roller worm culture fluid in flocculability inoculum or bioreactor for disposing polluted water, form flocculability antibacterial culturing system, described spinning roller worm culture fluid is as make consumption be flocculability antibacterial culturing system quality the 0.05～1% of flocculability bacterial growth promoter, by flocculability antibacterial culturing system is cultivated, namely obtain the culture fluid containing bacteria floccule, make flocculability antibacterial grow in flocculability antibacterial culturing system.Spinning roller worm is a kind of micro-metazoa common in activated sludge, belongs to Bdelloid rotifers section on taxonomy.In activated sludge process when the Bdelloid rotifers quantity such as spinning roller worm are more, there will be the quantity of the flocculability antibacterial phenomenon that substantially flocculability many, activated sludge is substantially good in activated sludge, the active substance that its reason can be secreted with spinning roller worm containing promoting flocculability bacterial growth is relevant.Therefore, the present invention promotes the growth of flocculability antibacterial by adding the spinning roller worm culture fluid of the active substance containing the secretion of spinning roller worm in activated sludge or flocculability bacteria culture media.
As currently preferred technical scheme, spinning roller worm culture fluid obtains through following steps:
A. spinning roller worm is inoculated into flour be bait distilled water or deionized water in, shading, shaking speed be 120rpm, 30 DEG C when cultivate spinning roller worm；
B., when the density after the spinning roller worm breeding cultivated in described step a reaches 100～500ind/mL, with 300 order nylon net filter spinning roller worm culture fluid, from spinning roller worm culture fluid, remove spinning roller worm, obtain first filtrate；
C. the ultrafilter membrane adopting aperture to be 0.2～0.5 μm, the first filtrate that described step b is obtained is filtered again, removes the fine particle thing in filtrate and other solid contents further, and collects filtrate, and the filtrate thus obtained is spinning roller worm culture fluid.The spinning roller worm culture fluid obtained in step c is added in bacteria culture media with the dosage that mass percent is 0.05 ~ 1%, is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 48 hours, namely obtain the culture fluid containing bacteria floccule.
As such scheme it is preferred that technical scheme, by adding spinning roller worm culture fluid in flocculability inoculum or bioreactor for disposing polluted water, form flocculability antibacterial culturing system, then flocculability antibacterial culturing system is placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 48 hours, namely obtain the culture fluid containing bacteria floccule.
Above-mentioned flocculability antibacterial is preferably the antibacterial of single kind, or is preferably the mixed cell of the antibacterial of the multiple types lived on activated sludge.
Above-mentioned flocculability antibacterial is preferably bioflocculant-producing bacteria.
Above-mentioned flocculability antibacterial is preferably the mixed vaccine of any one antibacterial in wax printing fabric, vesicle shortwave Zymomonas mobilis and bacillus thuringiensis or any several antibacterial.
The present invention compared with prior art, has following apparent prominent substantive distinguishing features and remarkable advantage:
1. the present invention utilizes the spinning roller worm culture fluid method to promote the growth of flocculability antibacterial, has green, environmental protection, non-secondary pollution, feature easy to use；
2. the present invention is under the effect of spinning roller worm culture fluid, and the growth of flocculability antibacterial obtains promotion, can make the more biological flocculant of flocculability bacterial secretory；
3. the spinning roller worm culture fluid of the present invention can promote the flocculability bacteria growing of multiple types, applied widely, can be effectively applied in activated sludge process.
Accompanying drawing explanation
Fig. 1 is that the embodiment of the present invention one adds spinning roller worm culture fluid and do not add the dry cell weight size comparison diagram of bacillus cereus during spinning roller worm culture fluid.
Fig. 2 is that the embodiment of the present invention two adds spinning roller worm culture fluid and do not add the dry cell weight size comparison diagram of vesicle shortwave Zymomonas mobilis during spinning roller worm culture fluid.
Fig. 3 is that the embodiment of the present invention three adds spinning roller worm culture fluid and do not add the dry cell weight size comparison diagram of mixed cell during spinning roller worm culture fluid.
Detailed description of the invention
Details are as follows for the preferred embodiments of the present invention:
Embodiment one:
In the present embodiment, referring to Fig. 1, utilize spinning roller worm culture fluid promote bacillus cereus (Bacilluscereus) method that grows, sequentially include the following steps:
A. the collecting cells of bacillus cereus: the bacillus cereus being stored in medium slant inoculating loop is seeded in LB culture medium, it is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 24 hours, centrifugal treating culture fluid, with deionized water wash, thus repeatedly after centrifugal, deionized water wash three times, with sterilized water, bacterial concentration is regulated to OD600Value 0.6, this solution is the bacteria suspension of bacillus cereus；
B. simulated domestic wastewater preparation: weigh 0.17g glucose, 0.16g starch, 0.233g sodium acetate, 0.025g ammonium chloride, 0.158g peptone, 0.04g Carnis Bovis seu Bubali cream, 0.0284g ammonium sulfate, 0.07g potassium dihydrogen phosphate and 0.06g sodium carbonate, dissolve in 1000mL deionized water；
C. the preparation of spinning roller worm culture fluid: spinning roller worm is inoculated into flour for the deionized water of bait, shading, shaking speed be 120rpm, at 30 DEG C when cultivate spinning roller worm 24h；When the density after the spinning roller worm breeding cultivated reaches 450ind/mL, with 300 order nylon net filter spinning roller worm culture fluid, separate from spinning roller worm culture fluid and remove spinning roller worm, obtain first filtrate, and repeatedly rinse the spinning roller worm being trapped on nylon wire 5 times with aseptic water, spinning roller worm is collected standby；The ultrafilter membrane adopting aperture to be 0.22 μm again, first filtrate is filtered again, removes the fine particle thing including antibacterial in filtrate and other solid contents further, and collect filtrate, filtrate is settled to 100mL, and the filtrate thus obtained is spinning roller worm culture fluid；
D. promote bacterial growth Preparatory work of experiment: two experimental grouies are set, one of which is matched group, another group is spinning roller worm culture fluid interpolation group, two experimental grouies all use 250mL conical beaker to be experiment container, with the bacillus cereus antibacterial that obtains in step a for experimental subject, with the simulated domestic wastewater that obtains in stepb for inoculum, the conical beaker of two experimental grouies adds the bacteria suspension of the simulated domestic wastewater of 90mL and the bacillus cereus of 10mL respectively, that adds 0.5mL in the conical beaker of interpolation group again prepares spinning roller worm culture fluid in step c；
E. antibacterial culturing: be placed on the shaking table of 120rpm by the conical beaker of obtain by step d two experimental grouies, in 30 DEG C of continuous cultivations 48 hours.
Bacterial cell dry weight is tested:
Adopt centrifuging to separate and collect the antibacterial of two experimental grouies that the present embodiment is cultivated, then pass through dry, the bacterial cell dry weight contrast obtaining two experimental grouies of weighing, as shown in Figure 1.
Spinning roller worm culture fluid promotes the growth result analysis of bacillus cereus:
As shown in Figure 1, inoculum with the addition of the interpolation group of spinning roller worm culture fluid, compared with the matched group being not added with spinning roller worm culture fluid, bacillus cereus dry cell weight adds 60%, it was shown that the growth of bacillus cereus is had obvious facilitation effect by spinning roller worm culture fluid.
The present embodiment by adding spinning roller worm culture fluid in flocculability inoculum or bioreactor for disposing polluted water, form flocculability antibacterial culturing system, then flocculability antibacterial culturing system is placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 48 hours, namely obtain the culture fluid containing bacteria floccule, make flocculability antibacterial grow in flocculability antibacterial culturing system.The present embodiment adopts the method that spinning roller worm culture fluid promotes flocculability bacterial growth, has ecology, easy to use, the obvious feature of effect.
Embodiment two:
The present embodiment is essentially identical with embodiment, is particular in that:
In the present embodiment, referring to Fig. 2, utilize spinning roller worm culture fluid promote vesicle shortwave Zymomonas mobilis (Brevundimonasvesicularis) method that grows, sequentially include the following steps:
A. the collecting cells of vesicle shortwave Zymomonas mobilis: the vesicle shortwave Zymomonas mobilis inoculating loop being stored in medium slant is seeded in LB culture medium, it is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 24 hours, centrifugal treating culture fluid, with deionized water wash, thus repeatedly after centrifugal, deionized water wash three times, with sterilized water, bacterial concentration is regulated to OD600Value 0.6, this solution is the bacteria suspension of vesicle shortwave Zymomonas mobilis；
B. simulated domestic wastewater preparation: this step is identical with the corresponding steps of embodiment one；
C. the preparation of spinning roller worm culture fluid: this step is identical with the corresponding steps of embodiment one；
D. promote bacterial growth Preparatory work of experiment: two experimental grouies are set, one of which is matched group, another group is spinning roller worm culture fluid interpolation group, two experimental grouies all use 250mL conical beaker to be experiment container, with the vesicle shortwave aeruginosa bacteria that obtains in step a for experimental subject, with the simulated domestic wastewater that obtains in stepb for inoculum, the conical beaker of two experimental grouies adds the simulated domestic wastewater of 90mL and the bacteria suspension of the vesicle shortwave Zymomonas mobilis of 10mL respectively, that adds 0.5mL in the conical beaker of interpolation group again prepares spinning roller worm culture fluid in step c；
E. antibacterial culturing: this step is identical with the corresponding steps of embodiment one.
Bacterial cell dry weight is tested:
Adopt centrifuging to separate and collect the antibacterial of two experimental grouies that the present embodiment is cultivated, then pass through dry, the bacterial cell dry weight contrast obtaining two experimental grouies of weighing, as shown in Figure 2.
Spinning roller worm culture fluid promotes the growth result analysis of vesicle shortwave Zymomonas mobilis:
As shown in Figure 2, inoculum with the addition of the interpolation group of spinning roller worm culture fluid, compared with the matched group being not added with spinning roller worm culture fluid, vesicle shortwave Pseudomonad cell dry weight adds 29.7%, it was shown that the growth of vesicle shortwave Zymomonas mobilis is had good facilitation effect by spinning roller worm culture fluid.
Embodiment three:
The present embodiment is essentially identical with embodiment, is particular in that:
In the present embodiment, referring to Fig. 3, utilize spinning roller worm culture fluid promote by bacillus cereus (Bacilluscereus), vesicle shortwave Zymomonas mobilis (Brevundimonasvesicularis), bacillus thuringiensis (Bacillusthuringiensis) these three antibacterial mix mixed vaccine growth method, sequentially include the following steps:
A. the collecting cells of mixed vaccine: bacillus cereus, vesicle shortwave Zymomonas mobilis, bacillus thuringiensis these three antibacterial are prepared into according to the method for the step a in embodiment one respectively the bacteria suspension of single bacterium, then the bacteria suspension of three kinds of bacterium is mixed into the ratio of 1:1:1 the bacteria suspension of mixed vaccine；
B. simulated domestic wastewater preparation: this step is identical with the corresponding steps of embodiment one；
C. the preparation of spinning roller worm culture fluid: this step is identical with the corresponding steps of embodiment one；
D. promote bacterial growth Preparatory work of experiment: two experimental grouies are set, one of which is matched group, another group is spinning roller worm culture fluid interpolation group, two experimental grouies all use 250mL conical beaker to be experiment container, with the mixed vaccine antibacterial that obtains in step a for experimental subject, with the simulated domestic wastewater that obtains in stepb for inoculum, adding the bacteria suspension of the simulated domestic wastewater of 90mL and the mixed vaccine of 10mL in the conical beaker of two experimental grouies respectively, that adds 0.5mL in the conical beaker of interpolation group again prepares spinning roller worm culture fluid in step c；
E. antibacterial culturing: this step is identical with the corresponding steps of embodiment one.
Bacterial cell dry weight is tested:
Adopt centrifuging to separate and collect the antibacterial of two experimental grouies that the present embodiment is cultivated, then pass through dry, the bacterial cell dry weight contrast obtaining two experimental grouies of weighing, as shown in Figure 3.
Spinning roller worm culture fluid promotes the growth result analysis of mixed vaccine:
From the figure 3, it may be seen that with the addition of the interpolation group of spinning roller worm culture fluid in inoculum, compared with the matched group being not added with spinning roller worm culture fluid, mixed vaccine dry cell weight adds 52.9%, it was shown that the growth of mixed vaccine is had good facilitation effect by spinning roller worm culture fluid.
Above in conjunction with accompanying drawing, the embodiment of the present invention is illustrated; but the invention is not restricted to above-described embodiment; multiple change can also be made according to the purpose of the innovation and creation of the present invention; change, modification, replacement, combination or the simplification made under all spirit according to technical solution of the present invention and principle; all should be the substitute mode of equivalence; as long as meeting the goal of the invention of the present invention; utilize know-why and the inventive concept of the method for spinning roller worm culture fluid promotion flocculability bacterial growth without departing from the present invention, broadly fall into protection scope of the present invention.

Claims (6)

1. one kind utilizes the method that spinning roller worm culture fluid promotes flocculability bacterial growth, it is characterized in that: by adding spinning roller worm culture fluid in flocculability inoculum or bioreactor for disposing polluted water, form flocculability antibacterial culturing system, described spinning roller worm culture fluid is as make consumption be flocculability antibacterial culturing system quality the 0.05～1% of flocculability bacterial growth promoter, by flocculability antibacterial culturing system is cultivated, namely obtain the culture fluid containing bacteria floccule, make flocculability antibacterial grow in flocculability antibacterial culturing system.
2. utilize the method that spinning roller worm culture fluid promotes flocculability bacterial growth according to claim 1, it is characterised in that described spinning roller worm culture fluid obtains through following steps:
A. spinning roller worm is inoculated into flour be bait distilled water or deionized water in, shading, shaking speed be 120rpm, 30 DEG C when cultivate spinning roller worm；
B., when the density after the spinning roller worm breeding cultivated in described step a reaches 100～500ind/mL, with 300 order nylon net filter spinning roller worm culture fluid, from spinning roller worm culture fluid, remove spinning roller worm, obtain first filtrate；
C. the ultrafilter membrane adopting aperture to be 0.2～0.5 μm, the first filtrate that described step b is obtained is filtered again, removes the fine particle thing in filtrate and other solid contents further, and collects filtrate, and the filtrate thus obtained is spinning roller worm culture fluid.
3. the method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth according to claim 1 or claim 2, it is characterized in that: by adding spinning roller worm culture fluid in flocculability inoculum or bioreactor for disposing polluted water, form flocculability antibacterial culturing system, then flocculability antibacterial culturing system is placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 48 hours, namely obtain the culture fluid containing bacteria floccule.
4. the method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth according to claim 1 or claim 2, it is characterised in that: described flocculability antibacterial is the antibacterial of single kind, or lives in the mixed cell of the antibacterial of multiple types on activated sludge.
5. utilize the method that spinning roller worm culture fluid promotes flocculability bacterial growth according to claim 4, it is characterised in that: described flocculability antibacterial is bioflocculant-producing bacteria.
6. utilize the method that spinning roller worm culture fluid promotes flocculability bacterial growth according to claim 5, it is characterised in that: described flocculability antibacterial is the mixed vaccine of any one antibacterial in wax printing fabric, vesicle shortwave Zymomonas mobilis and bacillus thuringiensis or any several antibacterial.
CN201610184834.1A 2016-03-29 2016-03-29 Promote the method for flocculability bacterial growth using spinning roller worm culture solution Expired - Fee Related CN105779345B (en)

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CN110894482A (en) * 2019-12-17 2020-03-20 上海大学 Method for promoting growth of flocculating bacteria

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