CN104805050A - Method for accelerating microbial fermentation by negative ion powder - Google Patents
Method for accelerating microbial fermentation by negative ion powder Download PDFInfo
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- CN104805050A CN104805050A CN201510085961.1A CN201510085961A CN104805050A CN 104805050 A CN104805050 A CN 104805050A CN 201510085961 A CN201510085961 A CN 201510085961A CN 104805050 A CN104805050 A CN 104805050A
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- 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
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- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
- C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
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- C12P7/649—Biodiesel, i.e. fatty acid alkyl esters
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- C12Y302/01001—Alpha-amylase (3.2.1.1)
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Abstract
The invention discloses a method for accelerating microbial fermentation by negative ion powder. The method comprises specific steps as follows: (1), a culture medium is put in a 1L triangular flask, bacillus subtilis or chlorella is inoculated and cultured for 1-3 days on a shaking bed at the temperature of 20-37 DEG C at a speed of 140-210 rpm, and a shake flask seed culture solution is obtained; (2), the culture medium and the negative ion powder are added proportionally to a fermentation tank and sterilized under the high pressure for 30 min, and the seed culture solution is inoculated to the fermentation tank; (3), the temperature of the tank is kept at 20 DEG C-37 DEG C, the tank pressure is 15-60 kap, the stirring speed is 80-130 rpm, the ventilation ratio is 1: (0.1-0.5), the fermentation culture time is 1-8 days, and bacterial strains and a fermentation liquid are filtered after being released from the tank. The negative ion powder capable of promoting rapid growth of microorganisms is added in a conventional fermentation tank, the growth speed of the microorganisms can be increased, accumulation of biomass and products is increased, the fermentation period is shortened, a buffer function is performed on pH of the solution, the quantity of demanded dissolved oxygen in the fermentation tank is guaranteed without increase of tank pressure or introduction of pure oxygen, and the negative ion powder can be recycled after fermentation ends.
Description
Technical field
The invention belongs to field of fermentation engineering, be specifically related to a kind of method that negative ion powder promotes fermentable.
Background technology
Along with the progress of human sciences, more and more deep to the research of microorganism, the effect that microorganism plays in the field such as industry, agricultural, medicine, food, the energy more and more attracts people's attention, the especially construction of recycling economy, be unable to do without the participation of microorganism and the development of utilisation technology thereof and innovation.Rumen microbial growth efficiency is high, energy consumption is low, produce biologically active substance useful in a large number, be with a wide range of applications and high economic worth, but when high density fermentation, improve the productive rate of final Fungal biodiversity and target product, shorten fermentation period, reduce water power and use the restraining factors remaining fermentable.
Most of microbe is all aerobic, comprise bacterium, actinomycetes, fungi and certain algae slightly, and microorganism can only utilize dissolved oxygen (DO), the very little gas-liquid mass transfer resistance that causes of the mass transfer coefficient of oxygen in water is large, so dissolved oxygen is in the first place of growth of aerobic microorganisms breeding.Under 25 DEG C, 1 normal atmosphere, the solubleness of oxygen in air in pure water is 0.25 mmol/L, solubleness in the fermented liquid containing a large amount of organic salt and inorganic salt only has 0.22 mmol/L, microorganism oxygen consumption rate is approximately 25 ~ 100 mmol/L/h, and when high density fermentation, oxygen consumption rate increases severely especially.Increase dissolved oxygen and need improve stirring velocity, the higher requirement to equipment of rotating speed is higher, the power of motor is enough large, tank body is enough stable, sealing replacement frequency can be higher, and a large amount of generation of bubble and the stream of defoamer add in tank, can affect the transmission of oxygen and nutraceutical mass transfer, and then suppress the growth of bacterium.At the growth period that microorganism is different, substratum carbon-nitrogen ratio is improper, dissolved oxygen is not enough, the existence of organic acid or alkaline matter all can cause the fluctuation of fermented liquid pH.The change of pH not only affects the form of microorganism, and changes membrane passage, the absorption of impeding nutritious substance and the secretion of meta-bolites.The important composition nucleic acid, protein etc. of body are more responsive to pH, along with the change of pH can suffer irreversible destruction.Suitable pH can stimulate microbial growth, improves biochemical reaction speed and the speed of growth of cell.In fermentation process, usually need to pass into oxygen and stream adds pH adjusting agent to maintain microbial growth environment, therefore, increase dissolved oxygen, research that maintenance system pH value is stabilized in high density fermentation is significant.
Negative ion powder receives the concern of people as a kind of new inorganic material with specific function, and its resource is very abundant.High reactivity negative ion powder can be formed tens thousand of to electrode, produces electrodispersion, can control the ph stability of water solution system in the moment of contact water to water; Due to piezoelectricity and pyroelectricity, negative ion powder is releasing far infrared when extraneous environmental change, makes the macromole group of water become subset, accelerate moisture movement, make ortho-water become active water, enhance the solvency power of water, seepage force, reach aqueous solution oxygen enrichment object.Therefore, negative ion powder is added in microbial fermentation system to the effect that can reach and increase dissolved oxygen and stable potential of hydrogen.
Summary of the invention
The object of this invention is to provide a kind of method that negative ion powder promotes fermentable, it is simple to operate, economically feasible.
The technical solution used in the present invention is as follows:
1, in 1L triangular flask, put into substratum, then by subtilis or chlorella access bottle, at 20 ~ 37 DEG C, 1 ~ 3d cultivated by the shaking table of 140 ~ 210rpm, obtains shake-flask seed nutrient solution.
2, in fermentor tank, add substratum and negative ion powder in proportion, autoclaving 30 min, and seed culture fluid is accessed in fermentor tank.
3, keep tank temperature 20 ~ 37 DEG C, tank pressure 15 ~ 60 kap, stirring velocity 80 ~ 130 rpm, air flow 1:0.1 ~ 0.5, fermented incubation time 1 ~ 8 d, then puts tank, filters bacterial strain and fermented liquid.
The described subtilis culture medium prescription put in triangular flask and fermentor tank is: peptone 10 g/L, yeast extract paste 5 g/L, sodium-chlor 10 g/L, water 1L, and adjust ph is 7.2.
The described chlorella culture medium prescription put in triangular flask and fermentor tank is: glucose 20 g/L, KNO
31.50 g/L, KH
2pO
41.25 g/L, MgSO
47 H
2o 1.25 g/L, FeSO
47H
2o 20 mg/L, A5 liquid microelement 1mg/L, water 1L, regulate pH value to be 7.
Described negative ion powder is the tourmaline powder that schorl is made through pulverizing, grinding, oven dry, cleaning, and its median size is 0.1 ~ 15 μm.
In described fermentor tank, the addition of negative ion powder is 0.1 ~ 5%(quality percent by volume of cultivating base unit weight).
The inoculum size of described seed liquor is 5% ~ 20%(percent by volume of fermentation liquid measure).
The present invention has obvious promoter action compared with the microorganism high density fermentation of routine, improve microorganism growth speed, increase biomass, shorten fermentation period, acid adding or alkali adjustment pH need not be flowed, increase tank pressure or pass into the demand that pure oxygen etc. ensures dissolved oxygen in fermentor tank, production cost reduces, energy efficient.Negative ion powder is reusable after fermentation ends in addition, does not produce secondary pollution.
Embodiment
Be below several specific embodiment of the present invention, further illustrate the present invention, but the present invention be not limited only to this.
Embodiment 1
In 1L triangular flask, add substratum, then subtilis is accessed in bottle, at 32 DEG C, the shaking table of 150 rpm is cultivated 36 h, obtain shake-flask seed nutrient solution.Substratum 7 L is added in proportion in 10 L fermentor tanks, in 1%(g/mL) ratio add particle diameter be 0.2 μm of negative ion powder, autoclaving 30 min, accesses in fermentor tank with the inoculum size accounting for fermented liquid 9% by seed culture fluid, with conventional subtilis high density fermentation for contrast.Adjust pH to 7.2, keep tank temperature 37 DEG C, tank pressure 30 kap, stirring velocity 100 rpm, air flow 1:0.3, fermented incubation time 32 h, fermentation time terminates to put tank, filters thalline and fermented liquid.Compare with control group, negative ion powder significantly improves the cellular form of subtilis, and cell size is better than control group, OD
600the logarithmic phase of pH-value determination pH result display subtilis shifts to an earlier date 4 ~ 7h, and stationary phase is long.Biomass comparatively control group improves 30% ~ 40%.The α-amylase of bacillus subtilis secretion is usually stable 5.5 ~ 8.0 time, pH increases or reduces all makes enzyme activity reduce, and negative ion powder stablizes the fluctuation of pH, and α-amylase significantly improves at the suitableeest scope inner enzyme vigor, compared with the control, enzyme activity improves 30 ~ 40%.
Embodiment 2
In 1L triangular flask, put into substratum, then subtilis is accessed in bottle, at 32 DEG C, the shaking table of 150 rpm is cultivated 36 h, obtain shake-flask seed nutrient solution.Substratum 7L is added in proportion in 10 L fermentor tanks, in 0.2%(g/mL) ratio add particle diameter be 15 μm of negative ion powders, autoclaving 30 min, accesses in fermentor tank with the inoculum size accounting for fermented liquid 9% by seed culture fluid, with conventional subtilis high density fermentation for contrast.Adjust pH to 7.2, keep tank temperature 37 DEG C, tank pressure 30 kap, stirring velocity 100 rpm, air flow 1:0.3, fermented incubation time 32 h, fermentation time terminates to put tank, filters thalline and fermented liquid.Compare with control group, negative ion powder significantly improves the cellular form of subtilis, and cell size is better than control group, OD
600the logarithmic phase of pH-value determination pH result display subtilis shifts to an earlier date 2 ~ 5h, and stationary phase is long.Biomass comparatively control group improves 20% ~ 32%.The α-amylase of bacillus subtilis secretion is usually stable 5.5 ~ 8.0 time, pH increases or reduces all makes enzyme activity reduce, and negative ion powder stablizes the fluctuation of pH, and α-amylase significantly improves at the suitableeest scope inner enzyme vigor, compared with the control, enzyme activity improves 25 ~ 30%.
Embodiment 3
In 1L triangular flask, put into substratum, then chlorella is accessed in bottle, at 25 DEG C, the shaking table of 180 rpm is cultivated 2 d, obtain shake-flask seed nutrient solution.Substratum 10L is added in proportion in 15 L fermentor tanks, in 1%(g/mL) ratio to add particle diameter be the negative ion powder of 0.1 μm, autoclaving 30 min, accesses in fermentor tank with the inoculum size accounting for fermented liquid 15% by seed culture fluid, with conventional chlorella high density fermentation for contrast.Adjust pH to 6.5, keep tank temperature 28 DEG C, tank pressure 30 kap, stirring velocity 100 rpm, air flow 1:0.4, fermented incubation time 7d, fermentation time terminates to put tank, filters frustule and fermented liquid.Get the oven dry of appropriate frustule to weigh, obtain biomass, extract biofuel in frustule by phenol chloroform method and detect its content.Result shows, in fermenting process, fermented liquid pH fluctuation range is little, and biomass and the biofuel content of chlorella significantly improve, and compared with control group, the increase rate of biomass and biofuel is respectively 25 ~ 40% and 15 ~ 25%.
Embodiment 4
In 1L triangular flask, put into substratum, then chlorella is accessed in bottle, at 25 DEG C, 2d cultivated by the shaking table of 180 rpm, obtain shake-flask seed nutrient solution.Substratum 10L is added in proportion in 15 L fermentor tanks, in 0.1%(g/mL) ratio add particle diameter be 12 μm of negative ion powders, autoclaving 30 min, accesses in fermentor tank with the inoculum size accounting for fermented liquid 15% by seed culture fluid, with conventional chlorella high density fermentation for contrast.Adjust pH to 6.5, keep tank temperature 28 DEG C, tank pressure 30 kap, stirring velocity 100 rpm, air flow 1:0.4, fermented incubation time 7 d, fermentation time terminates to put tank, filters frustule and fermented liquid.Get appropriate oven dry to weigh, obtain biomass, extract biofuel in frustule by phenol chloroform method and detect its content.Result shows, in fermenting process, fermented liquid pH fluctuation range is little, and negative ion powder improves biomass and the biofuel content of chlorella, and compared with control group, the increase rate of biomass and biofuel is respectively 15 ~ 25% and 6 ~ 15%.
Claims (5)
1. the method for a negative ion powder promotion fermentable, it is characterized in that concrete steps of the present invention are: (1) puts into substratum in 1L triangular flask, again by subtilis or chlorella access bottle, at 20 ~ 37 DEG C, 1 ~ 3d cultivated by the shaking table of 140 ~ 210rpm, obtains shake-flask seed nutrient solution;
(2) in fermentor tank, substratum and negative ion powder is added in proportion, autoclaving 30 min, and seed culture fluid is accessed in fermentor tank, (3) tank temperature 20 ~ 37 DEG C is kept, tank pressure 15 ~ 60 kap, stirring velocity 80 ~ 130 rpm, air flow 1:0.1 ~ 0.5, fermented incubation time 1 ~ 8 d, filters bacterial strain and fermented liquid after putting tank.
2. a kind of negative ion powder according to right 1 promotes the method for fermentable, it is characterized in that the described negative ion powder adding a certain amount of certain particle diameter in conventional microbiological high density fermentation, promotes the expression of microbial growth and product.
3. a kind of negative ion powder according to right 1 promotes the method for fermentable, and it is characterized in that described negative ion powder is that schorl passes through pulverizing, grinding, dries, cleans the tourmaline powder made, its median size is 0.1 ~ 15 μm.
4. a kind of negative ion powder according to right 1 promotes the method for fermentable, it is characterized in that the described negative ion powder addition added in fermentor tank is 0.1 ~ 5%(quality percent by volume of cultivating base unit weight).
5. a kind of negative ion powder according to right 1 promotes the method for fermentable, it is characterized in that the inoculum size of described seed culture fluid inoculation is 5% ~ 20%(percent by volume of fermentation liquid measure).
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106889307A (en) * | 2015-12-18 | 2017-06-27 | 顾国忠 | A kind of production technology of biological feedstuff of stalk |
| CN107712844A (en) * | 2017-10-13 | 2018-02-23 | 北京陨水生物科技有限公司 | A kind of preparation method and applications of meteoritic mineral ion |
| CN108285913A (en) * | 2017-12-09 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of technique preparing extraction L-Glutamine |
| CN108285914A (en) * | 2017-12-03 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of zymotechnique of L-Trp |
| CN109762857A (en) * | 2017-11-09 | 2019-05-17 | 卢松 | A kind of technique preparing xanthan gum |
| CN109797176A (en) * | 2017-11-17 | 2019-05-24 | 卢松 | A kind of environment-protective process preparing monosodium glutamate |
| CN109797175A (en) * | 2017-11-17 | 2019-05-24 | 卢松 | The fermentation separation method of sodium glutamate |
| CN110859862A (en) * | 2019-12-04 | 2020-03-06 | 杨大伟 | Method for improving effective components of angelica sinensis through microbial fermentation and transformation |
| CN115353998A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in composting and deodorizing of breeding houses |
| CN115353984A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in agricultural cultivation |
| CN115353999A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in garbage deodorization |
| CN115353985A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in agricultural planting |
| CN115627215A (en) * | 2022-12-08 | 2023-01-20 | 北京再益生物科技有限公司 | Efficiency of degradation is improved microbial fermentation jar |
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106889307A (en) * | 2015-12-18 | 2017-06-27 | 顾国忠 | A kind of production technology of biological feedstuff of stalk |
| CN107712844A (en) * | 2017-10-13 | 2018-02-23 | 北京陨水生物科技有限公司 | A kind of preparation method and applications of meteoritic mineral ion |
| CN109762857A (en) * | 2017-11-09 | 2019-05-17 | 卢松 | A kind of technique preparing xanthan gum |
| CN109797176A (en) * | 2017-11-17 | 2019-05-24 | 卢松 | A kind of environment-protective process preparing monosodium glutamate |
| CN109797175A (en) * | 2017-11-17 | 2019-05-24 | 卢松 | The fermentation separation method of sodium glutamate |
| CN108285914A (en) * | 2017-12-03 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of zymotechnique of L-Trp |
| CN108285913A (en) * | 2017-12-09 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of technique preparing extraction L-Glutamine |
| CN108285913B (en) * | 2017-12-09 | 2020-12-29 | 新疆阜丰生物科技有限公司 | Process for preparing and extracting L-glutamine |
| CN110859862A (en) * | 2019-12-04 | 2020-03-06 | 杨大伟 | Method for improving effective components of angelica sinensis through microbial fermentation and transformation |
| CN115353998A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in composting and deodorizing of breeding houses |
| CN115353984A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in agricultural cultivation |
| CN115353999A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in garbage deodorization |
| CN115353985A (en) * | 2022-08-30 | 2022-11-18 | 北京良土良生生物科技有限公司 | EM (effective microorganisms) culture method and application of fermentation product thereof in agricultural planting |
| CN115627215A (en) * | 2022-12-08 | 2023-01-20 | 北京再益生物科技有限公司 | Efficiency of degradation is improved microbial fermentation jar |
| CN115627215B (en) * | 2022-12-08 | 2023-04-28 | 北京再益生物科技有限公司 | Microbial fermentation tank capable of improving degradation efficiency |
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