CN103160453B - Preparation method of microecologics for nitrite degradation, microecologics and use thereof - Google Patents
Preparation method of microecologics for nitrite degradation, microecologics and use thereof Download PDFInfo
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- CN103160453B CN103160453B CN201310058985.9A CN201310058985A CN103160453B CN 103160453 B CN103160453 B CN 103160453B CN 201310058985 A CN201310058985 A CN 201310058985A CN 103160453 B CN103160453 B CN 103160453B
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- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 title claims abstract description 53
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- 235000018291 probiotics Nutrition 0.000 claims description 42
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- 230000000593 degrading effect Effects 0.000 claims description 25
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 20
- 230000008569 process Effects 0.000 claims description 15
- 239000002054 inoculum Substances 0.000 claims description 7
- 244000068988 Glycine max Species 0.000 claims description 6
- 235000010469 Glycine max Nutrition 0.000 claims description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
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- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 6
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims description 6
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Abstract
The invention discloses a preparation method of microecologics for nitrite degradation. The method of the microecologics for the nitrite degradation comprises the steps of preparing a seed solution of bacillus subtilis, enabling the seed solution to be inoculated into a liquid fermentation medium of a fermentation tank for fermental cultivation, adopting a two-step temperature control method during the fermental cultivation, adopting different temperatures to carry out fermentation in the earlier stage of the fermentation and in the later stage of the fermentation, after the fermentation is completed, concentrating a bacterium solution, drying thalli, diluting the thalli, detecting the thalli, packaging the thalli, and obtaining the microecologics of the degradation nitrite. Futher disclosed are the microecologics for the nitrite degradation and use of the microecologics for the nitrite degradation in aquaculture water quality improvement. The microecologics for the nitrite degradation is obtained through the method. According to the preparation method of the microecologics for the nitrite degradation, the two-step temperature control method is adopted, a viable count and an endospore rate are improved, and stability of product quality is ensured. Due to addition of zeolite powder in a liquid state fermentation media component, compared with a general immobilization method, the preparation method of the microecologics of the degradation nitrite achieves more effective immobilization, prolongs a quality guarantee period of a product, and an effect of the microecologics for the nitrite degradation is good.
Description
Technical field
The present invention relates to a kind of preparation method of probiotics of degrading nitrite, the invention still further relates to the probiotics that uses the degrading nitrite that makes of the method, the purposes of the probiotics that the present invention has also further related to this degrading nitrite in aquaculture water water quality improvement.
Background technology
Along with domestic and international culture fishery fast development, the expanding day of cultivation scale, improving constantly of intensive degree, the amount of transfiniting is put in a suitable place to breed and is concentrated bait throwing in to cause aquaculture water to produce a large amount of residual baits, ight soil and dead animals and plants corpse, these organic depositions are at the bottom of pond, putrid fermentation under the effect of heterotrophic bacterium, protein and nucleic acid wherein are slowly decomposed, produce a large amount of nitrogenous objectionable impuritiess, aquaculture water water quality is worsened rapidly, cause serious self-pollution, when serious, whole breeding ecological environment is all destroyed.Pollute while generation, ammonium nitrogen in water body, the hazardous and noxious substances such as nitrite produce in a large number, especially the nitrite of high density makes to cultivate the decline of object blood transport oxygen ability, impel the oxyphorase in blood to be converted into methemoglobin, lose the ability with oxygen combination, not only directly endanger cultivated animals, while is due to its long-term accumulate poisoning effect, cause fish, the disease resistances such as shrimp reduce, easily cause the invasion and attack of various pathogenic bacterias, therefore be considered to be fish, the pathogenic root of shrimp, the disease of bringing thus makes raiser suffer heavy losses, the pollution problem of water surrounding nitrite has become one of bottleneck of puzzlement culture fishery development, limit greatly the development of culture fishery.
The method of degrading nitrite mainly contains physics, chemistry and biological method in the market, employing is changed water, absorption, throwing and is executed the physico-chemical processes such as redox agent and carry out degrading nitrite, having the shortcomings such as consumption is large, the feature of environmental protection is poor, degradation rate is not high, it is short to hold time, easy bounce-back, is the behave of curing the symptoms, not the disease.And adopt the biological method degrading nitrite that adds probiotics have simple to operate, have no side effect, noresidue, effect is remarkable, cost is low, do not destroy many-sided advantages such as the eubiosis, has become the recent development trend of aquaculture water purification techniques.The outstanding effects of probiotics aspect degrading nitrite such as genus bacillus, photosynthetic bacterium, milk-acid bacteria constantly studies have reported that, wherein the advantage of subtilis, resistance to storage strong with its fecundity, is subject to extensive concern.
Existing probiotics product fermentation manufacturing technique mainly contains liquid-state fermentation technology and solid-state fermentation process, and liquid state fermentation product ubiquity living bacteria count and the low problem of gemma rate cause thalline in subsequent production treating processes to lose serious; The difficult monitoring of processing parameter and control in solid fermentation products production process, miscellaneous bacteria rate is high.In addition; after probiotics drops into aquaculture water, thalline is unbound state; tolerance to poor environment in water is low; be difficult to resist competition antagonism and the hydrobiological predation of indigenous bacterium in pond; and immobilization technology can play effective protection to the function yeast in probiotics; but existing immobilization embedded technological operation is loaded down with trivial details, is difficult to be applied to scale operation.Immobilization adsorption technology is simple to operate, carrier is with low cost, and current main processing mode is to drop into fixation support absorption by concentrated fermented liquid after fermentation ends again, between thalline and carrier a little less than bonding force, enter after water thalline and be easy to analyse and take off, do not reach effective immobilized object.
Although probiotics has many advantages in the application of degraded aquaculture system nitrite, but owing to there being above-mentioned many-sided defect, be developed so far, still exist in actual applications effect unstable or use the unfruitful problem of rear milli, substantially also do not have on the market can be really the effective specifics of degrading nitrite, cause nitrite pollution to become making the problem of aquaculture headache.
Summary of the invention
The object of the invention is to overcome the defect existing in prior art, provide that a kind of production cost is lower, simple to operate, the preparation method of the probiotics of product viable count and the high degrading nitrite of gemma rate.Another object of the present invention is to disclose the probiotics and the purposes of this probiotics in aquaculture water water quality improvement that use the prepared degrading nitrite of the inventive method.
For reaching described object, technical scheme of the present invention is as follows:
A preparation method for the probiotics of degrading nitrite, comprises the steps:
(a) prepare the seed liquor of subtilis (Bacillus subtilis).
(b) seed liquor of subtilis (Bacillus subtilis) is seeded in the liquid fermentation medium of fermentor tank and carries out fermentation culture, in fermentation culture, adopt two step method of temperature-control by, earlier fermentation and later stage are used different temperature to ferment, higher at earlier fermentation leavening temperature, lower in fermentation later stage fermentation temperature;
At earlier fermentation, thalline is early stage in lag phase and logarithmic growth, higher tank temperature is conducive to accelerate the sprouting of thalline, shorten time lag phase, in the fermentation later stage, thalline enters logarithmic growth vigorous period, and self metabolic heat production is serious, easily occur that local temperature is too high, suitably reduce control temperature more favourable with the raising of fermentative production bacterium number and the formation of later stage gemma.
(c) after having fermented, concentrated broth, thalline is dry, and dilution, detects, and packs, and obtains the probiotics of degrading nitrite.
Wherein, in above-mentioned steps (b), the inoculum size of subtilis (Bacillus subtilis) seed liquor is 5～10% of liquid fermentation medium weight; Described fermentation culture conditions is: Initial pH is 7～7.5, and rotating speed is 150～200r/min, and tank pressure is 0.04～0.06MPa, and fermentation time is 24～28h, and 35～38 DEG C of 1～16h temperature controls, 16h is to 28～32 DEG C of fermentation ends temperature controls; Described liquid state fermentation substratum is composed of the following components: note by weight percentage, Semen Maydis powder 2～3%, molasses 1～2%, soybean cake powder 1～2%, ammonium sulfate 0.2～0.5%, dipotassium hydrogen phosphate 0.1～0.2%, magnesium sulfate heptahydrate 0.03%, manganese sulfate monohydrate 0.01%, zeolite powder 5～10%, surplus is water.
Wherein, in above-mentioned steps (b), the inoculum size of subtilis (Bacillus subtilis) bacterium liquid is preferably 5% of liquid fermentation medium weight; Described fermentation culture conditions is preferably: Initial pH is 7～7.5, and rotating speed is 200r/min, and tank pressure is 0.04～0.06MPa, and fermentation time is 24h, and 37 DEG C of 1～16h temperature controls, 16h is to 32 DEG C of fermentation ends temperature controls; The each component concentration of described liquid state fermentation substratum is preferably: note by weight percentage, Semen Maydis powder 2%, molasses 1%, soybean cake powder 2%, ammonium sulfate 0.2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate heptahydrate 0.03%, manganese sulfate monohydrate 0.01%, zeolite powder 10%, surplus is water.
Wherein, in described step (c), the method for concentrated broth is centrifugal rear collection thalline; The dry method of thalline is conventional spray-drying process.
With aforesaid method obtain the probiotics of degrading nitrite and the purposes of the probiotics of described degrading nitrite in aquaculture water water quality improvement also belong to protection scope of the present invention.
The present invention compared with prior art, has the following advantages:
(1) adopt two step method of temperature-control by liquid fermentation method, fermented liquid viable count can reach 10,300,000,000/ml, and gemma rate more than 99%, has effectively improved viable count and gemma rate, has ensured the stability of quality product;
(2) in liquid state fermentation culture medium prescription, add zeolite powder, can make thalline in fermentation culture, enter in the microvoid structure of zeolite powder, in growing microorganism, realization and carrier combines closely, form firmly microbial film system, realize more firmly keying action than the conventional process for fixation that adds again zeolite powder after fermentation ends, realize effective immobilization, meanwhile, this operation has also extended the quality guaranteed period of probiotics;
(3) effect of gained probiotics degraded water nitrite is fine, and produces, uses simple to operate, with low cost, is easy to apply.
Detailed content of the present invention can obtain by explanation described later and institute's accompanying drawing.
Brief description of the drawings
Fig. 1 is the preparation method's of the probiotics of degrading nitrite of the present invention process route chart.
Fig. 2 is viable count situation of growth comparison in differing temps control mode bottom fermentation liquid.
Embodiment
Below in conjunction with drawings and Examples, the specific embodiment of the present invention is further described, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these amendments and replacement all fall within the scope of protection of the present invention.
Experiment material
The present invention subtilis used (Bacillus subtilis) is purchased from China Committee for Culture Collection of Microorganisms agricultural microorganism center, and production code member is: ACCC02973.
The preparation of preparation embodiment 1 liquid state fermentation substratum
Preparation liquid state fermentation substratum, it consists of: note by weight percentage, Semen Maydis powder 2%, molasses 1%, soybean cake powder 2%, ammonium sulfate 0.2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate heptahydrate 0.03%, manganese sulfate monohydrate 0.01%, zeolite powder 10%, surplus is water; Take by weight percentage each component and be added to the water, be stirred to evenly.
Preparation embodiment 2 does not contain the preparation of the liquid state fermentation substratum of zeolite powder
Preparation is not containing the liquid state fermentation substratum of zeolite powder, and it consists of: note by weight percentage, and Semen Maydis powder 2%, molasses 1%, soybean cake powder 2%, ammonium sulfate 0.2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate heptahydrate 0.03%, manganese sulfate monohydrate 0.01%, surplus is water; Take by weight percentage each component and be added to the water, be stirred to and be uniformly dissolved.
The preparation of the probiotics of embodiment 1 degrading nitrite
1. will buy subtilis (Bacillus subtilis) bacterial strain coming through PDA slant medium activation, choose bacterium and cultivate to being equipped with in the triangular flask of LB liquid nutrient medium, culture temperature is 37 DEG C, and incubation time is 24h; Cultured bacterium liquid in triangular flask is seeded to the seeding tank that LB liquid nutrient medium is housed again and expands fermentation culture, culture temperature is 37 DEG C, and incubation time is 24h, obtains the seed liquor of subtilis (Bacillus subtilis).
2. the seed liquor of step 1 gained is seeded in the liquid state fermentation substratum of fermentor tank and carries out fermentation culture, the inoculum size of seed liquor is 5% of liquid fermentation medium weight, fermentation culture conditions is: Initial pH is 7～7.5, rotating speed is 200r/min, tank pressure is 0.04～0.06MPa, fermentation time is 24h, and 37 DEG C of 1～16h temperature controls, 16h is to 32 DEG C of fermentation ends temperature controls.
3. after having fermented, by centrifugal fermented liquid liquid rear collection thalline, the thalline obtaining uses conventional spray-drying process to be dried, and being diluted to viable count is 21,000,000,000/g, detects, and packs, and obtains the probiotics of degrading nitrite.
The preparation of the probiotics of the rear immobilization degrading nitrite of reference examples 1
1. will buy subtilis (Bacillus subtilis) bacterial strain coming through PDA slant medium activation, choose bacterium and cultivate to being equipped with in the triangular flask of LB liquid nutrient medium, culture temperature is 37 DEG C, and incubation time is 24h; Cultured bacterium liquid in triangular flask is seeded to the seeding tank that LB liquid nutrient medium is housed again and expands fermentation culture, culture temperature is 37 DEG C, and incubation time is 24h, obtains the seed liquor of subtilis (Bacillus subtilis).
2. the seed liquor of step 1 gained is seeded to and in fermentor tank, carries out fermentation culture, fermentation culture is used the not liquid state fermentation substratum containing zeolite powder, the inoculum size of seed liquor is 5% of liquid fermentation medium weight, fermentation culture conditions is: Initial pH is 7～7.5, rotating speed is 200r/min, and tank pressure is 0.04～0.06MPa, and fermentation time is 24h, 37 DEG C of 1～16h temperature controls, 16h is to 32 DEG C of fermentation ends temperature controls.
3., after having fermented, in fermented liquid, add 10% zeolite powder absorption, by centrifugal fermented liquid rear collection thalline, the thalline obtaining uses conventional spray-drying process to be dried, and being diluted to viable count is 21,000,000,000/g, detects, pack, obtain the probiotics of rear immobilization degrading nitrite.
The preparation of the probiotics of reference examples 2 not immobilization degrading nitrites
1. will buy subtilis (Bacillus subtilis) bacterial strain coming through PDA slant medium activation, choose bacterium and cultivate to being equipped with in the triangular flask of LB liquid nutrient medium, culture temperature is 37 DEG C, and incubation time is 24h; Cultured bacterium liquid in triangular flask is seeded to the seeding tank that LB liquid nutrient medium is housed again and expands fermentation culture, culture temperature is 37 DEG C, and incubation time is 24h, obtains the seed liquor of subtilis (Bacillus subtilis).
2. the seed liquor of step 1 gained is seeded to and in fermentor tank, carries out fermentation culture, fermentation culture is used the not liquid state fermentation substratum containing zeolite powder, the inoculum size of seed liquor is 5% of liquid fermentation medium weight, fermentation culture conditions is: Initial pH is 7～7.5, rotating speed is 200r/min, and tank pressure is 0.04～0.06MPa, and fermentation time is 24h, 37 DEG C of 1～16h temperature controls, 16h is to 32 DEG C of fermentation ends temperature controls.
3. after having fermented, by centrifugal fermented liquid rear collection thalline, the thalline obtaining uses conventional spray-drying process to be dried, and being diluted to viable count is 21,000,000,000/g, detects, and packs, and obtains the probiotics of not immobilization degrading nitrite.
Test example 1 two step method of temperature-control by and step method of temperature-control by ferment effect contrast
1. test method
Prepare the seed liquor of subtilis according to step 1 described in embodiment 1, in step 2, three groups of tests are set and carry out ferment effect contrast, wherein:
Process A: 32 DEG C of omnidistance temperature controls ferment;
Treatments B: 37 DEG C of omnidistance temperature controls ferment;
Process C: adopt two step method of temperature-control by, temperature is carried out to segmentation control, 37 DEG C of front 16h temperature controls, after 16h, 32 DEG C of temperature controls.
In fermenting process, every 4h extracts the fermented liquid of every group and detects its viable count, the results are shown in Figure 2; The viable count and the gemma rate that when fermentation ends, detect every group of fermented liquid, the results are shown in Table 1.
2. test-results
Viable count situation of growth comparison in 2.1 fermenting processs
As can be seen from Figure 2, process the omnidistance temperature control of A at the lesser temps of 32 DEG C, early stage, bacterium number rose slowly, and the later stage enters that increased logarithmic phase bacterium number increases comparatively fast but be highly limited; The omnidistance temperature control of treatments B is at the comparatively high temps of 37 DEG C, and early stage, bacterium number rose soon, but fermentation later stage bacterium is counted rate of rise not as good as processing A and processing C; Process C and adopt two step method of temperature-control by segmentation control temperature to have a clear superiority in compared to processing A and treatments B, early stage, bacterium number rose soon, and the later stage can be realized the logarithmic growth of two-forty simultaneously.
Viable count and the comparison of gemma rate in 2.2 differing temps control mode bottom fermentation liquid
The comparison of viable count and gemma rate in table 1 differing temps control mode bottom fermentation liquid
| ? | A group (32 DEG C) | B group (37 DEG C) | C group (two step temperature controls) |
| Viable count (hundred million/mL) | 71 | 62 | 103 |
| Gemma rate (%) | 98.8 | 98.4 | 99.5 |
Table 1 data presentation, adopts bacterium number and gemma rate in two step temperature control method fermented liquids to reach respectively 10,300,000,000/mL and 99.5%, is all obviously better than the fermented liquid of the temperature control method gained of 32 DEG C of omnidistance fixed temperatures and 37 DEG C.The actual purifying water effect comparison of test example 2 different treatment probioticses
Tested on June 16,10 days～2011 June in 2011 and carry out in Jiangxia District, Wuhan City plant of many Foucaults skill farm.Polycultured ponds A, B and the C that 3 nitrite exceed standard, these 3 pond NO are selected in test
- 2-N concentration has all exceeded 0.15mg/L.Pond A is wherein done to throw the probiotics processing of executing the embodiment of the present invention 1; Pond B does to throw the probiotics processing of executing reference examples 1 of the present invention; Pond C does to throw the probiotics processing of executing reference examples 2 of the present invention; Each pond is thrown bacterium amount and is 200g/ mu rice.
After throwing bacterium on June 10, monitor continuously the nitrite content in 3 fishponds, the results are shown in Table 2
Table 2 nitrite exceed standard pond administer before and after nitrite content change (mg/L)
As can be seen from Table 2, the pond nitrite concentration that three kinds of probioticses have been executed in throwing has decline in various degree, wherein, the effect that embodiment 1 is executed in throwing is more remarkable compared with reference examples 1 and reference examples 2, its final degradation rate reaches 94.2%, apparently higher than rear immobilized 75.5% and not immobilized 58.2%, illustrate to adding zeolite powder in liquid state fermentation culture medium prescription and carry out front immobilization, than being fixed is not effective, realize more firmly keying action than the conventional process for fixation that adds again zeolite powder after fermentation ends, realize effective immobilization.
The quality guaranteed period comparison of test example 3 different treatment probioticses
By the 25 DEG C of preservations of constant temperature under air-proof condition of three kinds of probioticses of embodiment 1, reference examples 1 and reference examples 2 gained, regularly detect viable count, the results are shown in Table 3.
Viable count comparison in three kinds of probiotics preservation perives of table 3
Table 3 data presentation, in two years, the probiotics viable count storage rate of embodiment 1 reaches 91.6%, and the probiotics viable count storage rate of reference examples 1 is 43.8%, and the probiotics viable count storage rate of reference examples 2 is 27.3%.Result shows, the present invention carries out front immobilized mode to adding zeolite powder in liquid state fermentation culture medium prescription, and the survival of subtilis is had to provide protection, can effectively extend the preservation period of microbial inoculum.
Test example 4 product of the present invention purifying water effect in actual cultivating pool detects
This is tested on July 20,14 days～2011 July in 2011 and carries out in Jiangxia District, Wuhan City plant of many Foucaults skill farm.Grass carp cultivating pool A, B and the C that 3 nitrite exceed standard serious, these 3 pond NO are selected in test
- 2-N concentration has all exceeded 0.2mg/L, the aging blackout of Chi Shui, and ammonia nitrogen exceeds standard, and has large stretch of blue-green algae floating.Pond A is wherein done to throw the probiotics processing of executing the embodiment of the present invention 1, and throwing bacterium amount is 200g/ mu rice; Pond B does to throw and executes market like product microbial inoculum processing, and throwing bacterium amount is 200g/ mu rice; Pond C does not throw bacterium contrast.
After throwing bacterium on July 14, monitor continuously the nitrite content in 3 fishponds, the results are shown in Table 4:
Table 4 nitrite exceed standard pond administer before and after nitrite content change (mg/L)
As can be seen from Table 4, throw in the nitrite concentration of pond A after the probiotics of degrading cultivation water nitrite of the present invention and be obvious downtrending, throw bacterium after 3 days nitrite drop to 0.023mg/L by 0.281mg/L, clearance 91.8%, and do not have bounce-back at later stage pond nitrite concentration.The same period, two other pond nitrite concentration does not obviously decline, on the rise on the contrary.
Meanwhile, at duration of test, observe pond A water quality and obviously improve, water colour transfers to green refreshingly by black, and ammonia nitrogen is reduced to 0.31mg/L by 0.52mg/L after testing.
Claims (5)
1. a preparation method for the probiotics of degrading nitrite, comprises the steps:
(a) prepare the seed liquor of subtilis (Bacillus subtilis);
(b) seed liquor of subtilis (Bacillus subtilis) is seeded in the liquid fermentation medium of fermentor tank and carries out fermentation culture, described liquid fermentation medium is composed of the following components: by weight percentage, Semen Maydis powder 2～3%, molasses 1～2%, soybean cake powder 1～2%, ammonium sulfate 0.2～0.5%, dipotassium hydrogen phosphate 0.1～0.2%, magnesium sulfate heptahydrate 0.03%, manganese sulfate monohydrate 0.01%, zeolite powder 5～10%, surplus is water, in fermentation culture, adopt two step method of temperature-control by, fermentation time is 24h, to be 16h use different temperature to ferment to fermentation ends to earlier fermentation 1～16h and later stage, higher 37 DEG C at earlier fermentation leavening temperature, lower 32 DEG C in fermentation later stage fermentation temperature,
(c) after having fermented, concentrated broth, thalline is dry, and dilution, detects, and packs, and obtains the probiotics of degrading nitrite.
2. it is characterized in that in accordance with the method for claim 1:
In described step (b), the inoculum size of subtilis (Bacillus subtilis) seed liquor is 5～10% of liquid fermentation medium weight;
Described fermentation culture conditions is: Initial pH is 7～7.5, and rotating speed is 150～200r/min, and tank pressure is 0.04～0.06MPa.
3. it is characterized in that in accordance with the method for claim 1:
In described step (c), the method for concentrated broth is centrifugal rear collection thalline; The dry method of thalline is conventional spray-drying process.
4. it is characterized in that in accordance with the method for claim 2:
In described step (b), the inoculum size of subtilis (Bacillus subtilis) bacterium liquid is 5% of liquid fermentation medium weight;
Described fermentation culture conditions is: Initial pH is 7～7.5, and rotating speed is 200r/min, and tank pressure is 0.04～0.06MPa, and fermentation time is 24h, and 37 DEG C of 1～16h temperature controls, 16h is to 32 DEG C of fermentation ends temperature controls.
5. it is characterized in that in accordance with the method for claim 1:
In described step (b), liquid fermentation medium is composed of the following components: by weight percentage, Semen Maydis powder 2%, molasses 1%, soybean cake powder 2%, ammonium sulfate 0.2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate heptahydrate 0.03%, manganese sulfate monohydrate 0.01%, zeolite powder 10%, surplus is water.
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| CN105884040A (en) * | 2016-05-11 | 2016-08-24 | 长沙学院 | Preparation method of water body denitrification material |
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