CN103911324B - Contain probiotics preparation of Lactobacillus plantarum and bacillus licheniformis and preparation method thereof - Google Patents
Contain probiotics preparation of Lactobacillus plantarum and bacillus licheniformis and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to probiotics fermention technical field, be specifically related to a kind of taking dregs of beans, wheat bran and corn flour as matrix, with probio Lactobacillus plantarum HM-10 (Lactobacillus? plantarum? and bacillus licheniformis BL-09 (Bacilluslicheniformis HM-10)? BL-09) be the mixed solid fermentation technology of bacterial classification, this technology can be widely used in the aspects such as animal microecological. The method can be produced the fermented type feed with special sour fragrance, and the viable count of fermented product is high, and the degradation rate of dregs of beans protein is high. This preparation method's production cost is low, can increase economic efficiency and simple to operate.
Description
Technical field
The invention belongs to probiotics fermention technical field, be specifically related to a kind of taking dregs of beans, wheat bran and corn flour as matrix, with prebioticBacterium Lactobacillus plantarum HM-10 (LactobacillusplantarumHM-10) and bacillus licheniformis BL-09 (BacillusLicheniformisBL-09) be the mixed solid fermentation technology of bacterial classification, this technology can be widely used in the aspects such as animal microecological.
Background technology
At present, probiotic composition adopt more probio directly with liquid cow's milk and soya-bean milk fermentation, obtain functional yoghurt orFermented Soybean Milk, all the other adopt liquid state fermentation, directly collect probio thalline, are made into probio pulvis, tablet or capsule.Adopt liquid state fermentation meeting to produce a large amount of waste water, environmental pollution is larger, and post processing is more difficult, machinery equipment complex operation.Therefore the application adopts Lactobacillus plantarum HM-10 and bacillus licheniformis BL-09 mixed solid fermentation dregs of beans, wheat bran and corn flour,Adopt solid state fermentation, its production cost is low, and the fund of investment is less, convenient in downstream, pollutes littlely, device structure is simple,Easy to operate. Adopt Lactobacillus plantarum HM-10 and bacillus licheniformis BL-09 to carry out mixed culture solid state fermentation, can produce one and haveThe fermented product of acid fragrance taste, improves the palatability of animal feed. Its protein is broken down into little peptide and free amino acid, favourableAbsorb in animal intestine peptic digest.
Patent " a kind of effective prevention and improve the compound lactobacillus probiotics of garget ", CN103202391A, openThe compound micro-ecological preparation that contains Lactobacillus casei HM-09, Lactobacillus plantarum HM-10. Described Lactobacillus casei and plant breast barBacterium has been great Tun road, Beijing in address on October 29th, 2012, China Committee for Culture Collection of Microorganisms's common micro-organismsCenter preservation, Lactobacillus casei HM-09 (L.caseiHM-09) preserving number: CGMCCNo.6736; Lactobacillus plantarum HM-10(L.plantarumHM-10) preserving number: CGMCCNo.6741.
Summary of the invention
The object of the invention is to adopt Lactobacillus plantarum HM-10 (LactobacillusplantarumHM-10) and lichens gemma barBacterium BL-09(BacilluslicheniformisBL-09) carry out mixed solid fermentation dregs of beans, wheat bran and corn flour, provide onePlant the probio solid-fermented technique that can be widely used in micro-ecosphere taking dregs of beans, wheat bran and corn flour as matrix, the method canTo produce the fermented type feed with special sour fragrance, and the viable count of fermented product is high, and the degradation rate of dregs of beans protein is high.This preparation method's production cost is low, can increase economic efficiency and simple to operate.
The present invention seeks to be achieved through the following technical solutions.
Can be widely used in a probio solid-fermented technique for micro-ecosphere taking dregs of beans, wheat bran and corn flour as matrix, bagDraw together cultivation and the mixed solid fermentation thereof of Lactobacillus plantarum HM-10 and bacillus licheniformis BL-09, obtain the solid state fermentation of probioProduct.
Described mixed solid fermentation condition is: first with 1-5% inoculum concentration (v/w), Lactobacillus plantarum HM-10 is inoculated in to solid state fermentation trainingSupport base, temperature control 28-32 DEG C, relative humidity 50%-60%; After fermentation 0-12, access bacillus licheniformis BL-09 with 2-8% inoculum concentrationCarry out mixed solid fermentation, temperature control 34-37 DEG C, relative humidity 50-60%, continues fermentation 24-36h.
Mixed fermented culture can obtain product through pulverizing packing.
Solid-state fermentation culture medium composition comprises dregs of beans, wheat bran and corn flour, and the mass ratio of described dregs of beans, wheat bran and corn flour is:Dregs of beans, wheat bran, corn flour=10:8-10:0.5-1, water content is controlled at 40-75%.
After Lactobacillus plantarum HM-10 and bacillus licheniformis BL-09 mixed fungus fermentation finish, under optimum condition, free ammonical nitrogenContent can reach 800.1 μ mol/g, before fermentation, improves approximately 8 times, and bacillus licheniformis BL-09 gemma rate ferments more separately 1.5Doubly. The number of live bacteria of probiotics of mixed culture fermentation product is 1010More than cfu/g, wherein Lactobacillus plantarum HM-10 viable count reaches3.50×109Cfu/g is above, bacillus licheniformis BL-09 viable count reaches 1.50 × 1010More than cfu/g.
Above-mentioned mixed culture fermentation probiotic products can be applicable to the fields such as feed addictive.
Described Lactobacillus plantarum HM-10 and bacillus licheniformis BL-09 pass through respectively seed culture before mixed solid fermentation. SeedCulture medium is (g/L): peptone 10-20, and glucose 10-20, all the other are water, pH7.0-7.2. Bacillus licheniformis BL-09Seed culture condition: shaking speed is 150-180r/min, and temperature is 30-37 DEG C, cultivate 20-24h. Described plant breastThe seed culture condition of bacillus HM-10, is placed in constant incubator and leaves standstill cultivation, cultivation temperature 30-37 DEG C, and incubation time is 20-24h.
It is common that described bacillus licheniformis BL-09 is preserved in China Committee for Culture Collection of Microorganisms on December 31st, 2011Microorganism center, preserving number is CGMCCNo.5686, systematic name: bacillus licheniformis (Bacilluslicheniformis),Preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101.
Described bacillus licheniformis BL-09 is that a strain separates in healthy Roll road, area, Hohhot City ,Inner Mongolia Autonomous Region sampleBacillus, belong to amphimicrobian, Gram-positive bacillus. Thalline be rod shaft-like, be good at once in a while shaft-like, gemmaMiddle life, ovalize. Optimum growth temperature is 37 ± 1 DEG C, and the most suitable growth pH is 7.0-7.4, and solid state fermentation is dried product normal temperatureStore 6 months, gemma loss late is cultivated 12h left and right lower than 5%. nutrient agar panels, and bacterium colony is irregular, edge is rough, bacteriumThe central authorities that fall have obvious protruding peak; Be cultured to 24-36h, in the obvious and fold of bacterium colony fold, material is liquid, and oese is chosenGet and have wire drawing situation, microscopy gemma comes off substantially. Determine through uncle's Jie Shi bacterium handbook (the 9th edition) and bacterium, colonial morphologyFor bacillus licheniformis, called after bacillus licheniformis BL-09. Experiment in vitro confirms, this bacterial strain has good high temperature resistant, resistance toThe biochemical characteristics such as acid, bile tolerance, can also high proteinase yield, the multiple enzyme such as amylase, lipase and amino acid, vitamin,The several physiological active substances such as antibacterial material, the solid medium viscosity after fermentation is very large, and wire drawing is serious, and culture vessel bottom canSee a small amount of liquid substance, prove that the moisture-retaining capacity in its solid state fermentation is relatively strong.
Acid resistance test:
Bacillus licheniformis BL-09 is in meat soup seed liquor, and 37 DEG C, 180r/min cultivates 24h, in 4 DEG C, 4000r/minCentrifugal 10min collects thalline, with after 0.85% SPSS rinsing once, is suspended in pH and is respectively 2.0,3.0 and 4.0In cushioning liquid, cultivate 1-3h for 37 DEG C, detect viable count and change, it is good resistance to that result shows that bacillus licheniformis BL-09 hasAcid.
Heat resistant test:
Bacillus licheniformis BL-09 is in meat soup seed liquor, and 37 DEG C, 180r/min cultivates 24h, in 4 DEG C, 4000r/minCentrifugal 10min collects thalline, and with after 0.85% SPSS rinsing once, the thalline of acquisition is rare with appropriate SPSSAfter releasing, process 10min to obtain gemma in 80 DEG C of heating water baths, then process respectively 5-30 at 80 DEG C, 90 DEG C and 100 DEG CMin, detects viable count and changes, and result shows that bacillus licheniformis BL-09 has good heat resistance.
Bile tolerance test:
Bacillus licheniformis BL-09 is in meat soup seed liquor, and 37 DEG C, 180r/min cultivates 24h, in 4 DEG C, 4000r/minCentrifugal 10min collects thalline, with after 0.85% SPSS rinsing once, be suspended in containing gallbladder salinity be respectively 0.03%,0.10%, in 0.30% physiological saline, cultivate 1-6h for 37 DEG C, detect viable count and change, result shows bacillus licheniformis BL-09There is good bile tolerance.
Beneficial effect:
1. in the present invention, adopt Lactobacillus plantarum HM-10 and bacillus licheniformis BL-09 to carry out mixed solid fermentation dregs of beans, utilizeBacillus licheniformis BL-09 bacterial strain advantage is also explored optimization of fermentation conditions, has that production cost is low, fund input is few, downstream partManage easy, pollute the advantages such as little, device structure is simple to operation;
2. adopt Lactobacillus plantarum HM-10 to ferment, can produce the tart flavour such as lactic acid and acetic acid small-molecule substance, these little pointSub-material is given improvement and the variation of the organoleptic features such as the distinctive sour fragrance of product, color and luster and mouthfeel, makes fermented product easierBy animal is digested and assimilated; Aspect the mechanism of action, the gemma of bacillus licheniformis can be enter can be rapid after host's enteron aisleSprout, breed, consume oxygen in intestines, suppress harmful growth that needs bacterium, and protect, promote the growth of probio, to safeguardIntestinal microecology balance. Lactobacillus plantarum can surely be grown in host's enteron aisle and can utilize lactose to produce lactic acid, effectively alleviates AsiaPeople's lactose intolerance symptom. The lactic acid energy regulating intestinal canal pH producing, kills not acid resistance pathogen, and the hydrogen peroxide producing,Bacteriocins etc., have significant inhibitory action to other cause of diseases or putrefactive microorganisms, and on probio without impact, thereby maintain intestinesRoad colony balance; Secretion that can induction of immunity globulin A, improves host's immunologic function, can pass through the adjusting to interleukinAnd the symptom of alleviation colitis. The present invention obtain bacillus licheniformis and Lactobacillus plantarum composite probiotics preparations with more thanFunction, has good application development prospect.
3. adopt bacillus licheniformis BL-09 fermented bean dregs, wheat bran and corn flour, can make the protein in dregs of beans and corn flourBe broken down into little peptide and free amino acid, be beneficial to digesting and assimilating of stomach. Bacillus licheniformis BL-09 have good high temperature resistant,Biochemical characteristics such as acidproof, bile tolerance and produce the features such as multiple enzyme such as protease, amylase, lipase, exist product of the present inventionIn feed, add while use, this series products can effectively improve the utilization ratio 15-28% of feed while being used for Cow-feeding. The present invention producesThe addition of product in feed is 20-50 gram/kg. After product of the present invention uses, the conception rate of milk cow has improved approximately 12%, as calculatedCan improve the year output of milk more than approximately 300 kilograms of milk cow.
Detailed description of the invention
Below in conjunction with example, the present invention is further described, and following example is illustrative, is not determinate, can not be with followingExample limits protection scope of the present invention.
Embodiment 1: the mixed solid fermentation of probiotics preparation
(1) seed liquor of thalline preparation
Bacillus licheniformis BL-09 is inoculated in liquid seed culture medium from the glycerine pipe of cold storage, is positioned on shaking table, and rotating speed is150-180r/min, temperature is 30-37 DEG C, cultivates 20-24h. Lactobacillus plantarum HM-10 is inoculated in liquid seed culture medium,Be placed in constant incubator and leave standstill cultivation, cultivation temperature 30-37 DEG C, incubation time is 20-24h. Wherein liquid seed culture medium is(g/L): peptone 10-20, glucose 10-20, all the other are water, pH7.0-7.2.
(2) solid state fermentation
First with 1-5% inoculum concentration (v/w), Lactobacillus plantarum HM-10 is inoculated in to solid-state fermentation culture medium, temperature control 28-32 DEG C is relatively wetDegree 50%-60%; After fermentation 0-12, carry out mixed solid fermentation, temperature control 34-37 with 2-8% inoculum concentration access bacillus licheniformis BL-09DEG C, relative humidity 50-60%, continues fermentation 24-36h.
Solid-state fermentation culture medium composition comprises dregs of beans, wheat bran and corn flour, and the mass ratio of described dregs of beans, wheat bran and corn flour is:Dregs of beans, wheat bran, corn flour=10:8-10:0.5-1, water content is controlled at 40-75%.
(3) measure viable count
The general colony counting method that adopts, operating procedure is: take the bean pulp fermentation goods 10g preparing, be placed in the aseptic of 90mlIn physiological saline, be placed on shaking table and shake 30min, shake up and add the aseptic of another 9ml with the bacterium liquid that liquid-transfering gun is got 1ml afterwardsIn physiological saline, dilute, according to national standard method, take turns doing ten times of gradient dilutions. After having diluted, get respectively with liquid-transfering gunSuitable dilution factor 1ml throws flat board into, pours culture medium into and mixes, take turns doing three parallel, be placed in incubator 37 DEG C leave standstillCultivate 48h.
(4) mensuration of amino-acid nitrogen content
Get the product after 10g fermentation, use homogenizer fragmentation, mix with the distilled water of 90ml afterwards, place 1 day at 4 DEG C of refrigerators.The centrifugal 7000r/min of sample, centrifugal 15min. Supernatant 0.8ml is placed in test tube, then adds 3.2ml distilled water, 2.0mlDeveloper, mixes, and puts into boiling water bath and heats 15min, makes blank simultaneously. Then cold water is cooling, adds 5.0ml40%Ethanolic solution mixes, and after placement 15min, uses 1cm cuvette, returns to zero and measures A value in 570nm place with blank tube.
Embodiment 2: the impact of Lactobacillus plantarum HM-10 inoculum concentration on viable count
Lactobacillus plantarum HM-10 is inoculated in liquid seed culture medium, and be placed in constant incubator and leave standstill cultivation, 37 DEG C of cultivation temperature,Incubation time is 20h. Wherein liquid seed culture medium mass percent consists of (g/L): peptone 10, and glucose 20,All the other are water, pH7.0-7.2.
Solid-state fermentation culture medium consists of dregs of beans, wheat bran and corn flour, and the mass ratio of described dregs of beans, wheat bran and corn flour is:Dregs of beans, wheat bran, corn flour=10:10:0.5. Water content is controlled at 70%, 115 DEG C of sterilizing 20min.
In solid medium, inoculate respectively 1%(v/w), 3%(v/w), 5%(v/w) Lactobacillus plantarum HM-10, single bacterium sends outAfter ferment 48h, record Lactobacillus plantarum HM-10 viable count in tunning and be respectively 3.01 × 109cfu/g、2.97×109cfu/g、2.91×109Cfu/g. So determine that the optimum inoculation amount of Lactobacillus plantarum HM-10 is 1%(v/w).
Result: the suitableeest inoculum concentration of Lactobacillus plantarum HM-10 is 1%(v/w).
Embodiment 3: the impact of bacillus licheniformis BL-09 inoculum concentration on viable count
Bacillus licheniformis BL-09 is inoculated in liquid seed culture medium from the glycerine pipe of cold storage, is positioned on shaking table, and rotating speed is180r/min, temperature is 37 DEG C, cultivates 20h. Lactobacillus plantarum HM-10 is inoculated in liquid seed culture medium, is placed in constant temperatureIncubator leaves standstill to be cultivated, 37 DEG C of cultivation temperature, and incubation time is 20h. Wherein liquid seed culture medium mass percent consists of(g/L): peptone 10, glucose 20, all the other are water, pH7.0-7.2.
Solid-state fermentation culture medium consists of dregs of beans, wheat bran and corn flour, and the mass ratio of described dregs of beans, wheat bran and corn flour is:Dregs of beans, wheat bran, corn flour=10:10:0.5. Water content is controlled at 60%, 115 DEG C of sterilizing 20min.
In solid state fermentation, inoculate 1%(v/w) Lactobacillus plantarum HM-10,30 DEG C of temperature controls, relative humidity 60%; Fermentation 12h after againInoculate respectively 2%(v/w), 5%(v/w), 8%(v/w) bacillus licheniformis BL-09 carry out mixed solid fermentation, temperature control37 DEG C, relative humidity 60%, then continue fermentation 36h.
When the inoculum concentration of bacillus licheniformis BL-09 is 2%, record mixed culture fermentation product bacillus licheniformis BL-09 viable count and be8.11×109Cfu/g, Lactobacillus plantarum HM-10 viable count is 2.51 × 109Cfu/g, measuring free ammonical nitrogen content can reach742.6μmol/g。
When the inoculum concentration of bacillus licheniformis BL-09 is 5%, record bacillus licheniformis BL-09 viable count in mixed culture fermentation product and be1.05×1010Cfu/g, Lactobacillus plantarum HM-10 viable count is 3.21 × 109Cfu/g, measuring free ammonical nitrogen content can reach800.1μmol/g。
When the inoculum concentration of bacillus licheniformis BL-09 is 8%, record bacillus licheniformis BL-09 viable count in mixed culture fermentation product and be9.04×1010Cfu/g, Lactobacillus plantarum HM-10 viable count is 2.71 × 109Cfu/g, measuring free ammonical nitrogen content can reach773.4μmol/g。
Conclusion: the suitableeest inoculum concentration of bacillus licheniformis BL-09 is 5%(v/w).
Embodiment 4: the impact of fermentation time on mixed culture fermentation
Bacillus licheniformis BL-09 is inoculated in liquid seed culture medium from the glycerine pipe of cold storage, is positioned on shaking table, and rotating speed is180r/min, temperature is 37 DEG C, cultivates 20h. Lactobacillus casei HM-09 is inoculated into respectively in liquid seed culture medium, is placed inConstant incubator leaves standstill to be cultivated, 37 DEG C of cultivation temperature, and incubation time is 20h. Wherein liquid seed culture medium mass percent groupBecome (g/L): peptone 10, glucose 20, all the other are water, pH7.0-7.2.
Solid-state fermentation culture medium consists of dregs of beans, wheat bran and corn flour, and the mass ratio of described dregs of beans, wheat bran and corn flour is:Dregs of beans, wheat bran, corn flour=10:9:1. Water content is controlled at 55%, 115 DEG C of sterilizing 20min.
In solid-state fermentation culture medium, inoculate 1%(v/w) Lactobacillus casei HM-09,30 DEG C of temperature controls, relative humidity 55%, fermentationAfter 12h, inoculate 5%(v/w) bacillus licheniformis BL-09,35 DEG C of temperature controls, relative humidity 55%, then mixed fungus fermentation 24h.Recording bacillus licheniformis BL-09 viable count is 6.11 × 109Cfu/g, Lactobacillus casei HM-09 viable count is 2.10 × 109cfu/g，Free ammonical nitrogen content can reach 685.3 μ mol/g.
In solid-state fermentation culture medium, inoculate 1%(v/w simultaneously) Lactobacillus casei HM-09 and 5%(v/w) bacillus licheniformisBL-09,35 DEG C of temperature controls, relative humidity 55%, mixed fungus fermentation 48h. Record bacillus licheniformis BL-09 viable count and be 4.23 ×109Cfu/g, Lactobacillus casei HM-09 viable count is 3.68 × 109cfu/g, free ammonical nitrogen content can reach 731.0 μ mol/g.In solid-state fermentation culture medium, inoculate 1%(v/w) Lactobacillus casei HM-09,30 DEG C of temperature controls, relative humidity 40%, mixed fungus fermentationAfter 12h, inoculate 5%(v/w) bacillus licheniformis BL-09,35 DEG C of temperature controls, relative humidity 55%, then continue mixed bacterium extremely36h. Recording bacillus licheniformis BL-09 viable count is 9.20 × 109Cfu/g, Lactobacillus casei HM-09 viable count is 3.33 × 109Cfu/g, free ammonical nitrogen content can reach 778.5 μ mol/g.
Claims (6)
1. a composite probiotics ferment product, is sent out through hybrid solid-state by Lactobacillus plantarum CGMCCNo.6741 and bacillus licheniformisFerment makes, and it is characterized in that, the deposit number of described bacillus licheniformis is CGMCCNo.5686; Described mixed solid fermentation bagDraw together: first with 1-5%v/w inoculum concentration, Lactobacillus plantarum CGMCCNo.6741 is inoculated in to solid-state fermentation culture medium, temperature control 28-32 DEG C,Relative humidity 50%-60%, carries out hybrid solid-state with 2-8% inoculum concentration access bacillus licheniformis CGMCCNo.5686 after fermentation 12h and sends outFerment, temperature control 34-37 DEG C, relative humidity 50-60%, continues fermentation 24-36h; Described solid-state fermentation culture medium composition comprise dregs of beans,Wheat bran and corn flour; Described composite probiotics ferment product free ammonia ground state nitrogen content reaches 800.1 μ mol/g, and probio livesBacterium number is 1010More than cfu/g, wherein Lactobacillus plantarum viable count reaches 3.50 × 109Cfu/g, bacillus licheniformis viable count reaches1.50×1010cfu/g。
2. composite probiotics ferment product as claimed in claim 1, is characterized in that, the matter of described dregs of beans, wheat bran and corn flourAmount ratio is: dregs of beans, wheat bran, corn flour=10:8-10:0.5-1, water content is controlled at 40-75%.
3. composite probiotics ferment product as claimed in claim 1 or 2, is characterized in that, described Lactobacillus plantarum and lichens budSpore bacillus passes through respectively seed culture before mixed solid fermentation, and seed culture medium is: peptone 10-20g/L, glucose10-20g/L, all the other are water, pH7.0-7.2; The seed culture condition of bacillus licheniformis: 150-180r/min, temperature is30-37 DEG C, cultivates 20-24h; The seed culture condition of Lactobacillus plantarum: be placed in constant incubator and leave standstill cultivation, cultivation temperature30-37 DEG C, incubation time is 20-24h.
4. the process for solid state fermentation of the arbitrary described composite probiotics ferment product of claim 1-3, is characterized in that described methodComprise the steps: that Lactobacillus plantarum CGMCCNo.6741 and bacillus licheniformis CGMCCNo.5686 pass through respectively seed culturePrepare microbial inoculum; Mixed solid fermentation: first with 1-5%v/w inoculum concentration, Lactobacillus plantarum is inoculated in to solid-state fermentation culture medium, temperature control28-32 DEG C, relative humidity 50%-60%, carries out mixed solid fermentation with 2-8% inoculum concentration access bacillus licheniformis after fermentation 12h,Temperature control 34-37 DEG C, relative humidity 50-60%, continues fermentation 24-36h; Mixed fermented culture can obtain product through pulverizing packing.
5. the application of the arbitrary described composite probiotics ferment product of claim 1-3 in feed addictive.
6. a strain has bacillus licheniformis (Bacilluslicheniformis) CGMCC high temperature resistant, acidproof, bile tolerance characteristicNo.5686。
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| CN104472947A (en) * | 2014-12-15 | 2015-04-01 | 新疆农业大学 | Method suitable for feeding mares in lactation by supplementing microbial preparations |
| CN106720342A (en) * | 2017-01-23 | 2017-05-31 | 合肥工业大学 | A kind of preparation method of flammulina velutipes yoghourt |
| CN107006677A (en) * | 2017-04-27 | 2017-08-04 | 齐齐哈尔大学 | A kind of feed and its application rich in probiotics and active peptide |
| CN108552417A (en) * | 2018-04-12 | 2018-09-21 | 宁夏犇元生物科技有限公司 | A kind of compound probiotic functional feed and preparation method thereof |
| CN109576192A (en) * | 2019-01-14 | 2019-04-05 | 湖北华扬科技发展有限公司 | A kind of probiotics preparation, preparation method and applications |
| CN110982751B (en) * | 2019-12-27 | 2021-07-27 | 天津开发区坤禾生物技术有限公司 | Microbial compound microbial agent, microencapsulated compound microbial ecological agent, meat rabbit feed, and preparation method and application thereof |
| CN112980740B (en) * | 2021-04-12 | 2021-09-28 | 江苏省农业科学院 | Bacillus licheniformis and application thereof |
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