AU2016297402B2 - Bacillus coagulans, high cell density fermentation method thereof, and method for preparing spray-dried powder thereof - Google Patents

Bacillus coagulans, high cell density fermentation method thereof, and method for preparing spray-dried powder thereof Download PDF

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AU2016297402B2
AU2016297402B2 AU2016297402A AU2016297402A AU2016297402B2 AU 2016297402 B2 AU2016297402 B2 AU 2016297402B2 AU 2016297402 A AU2016297402 A AU 2016297402A AU 2016297402 A AU2016297402 A AU 2016297402A AU 2016297402 B2 AU2016297402 B2 AU 2016297402B2
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fermentation
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bacillus coagulans
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Liang Zhang
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Jiangnan University
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    • C12N1/205
    • C12R2001/07
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor

Abstract

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Description

Description
Title of the Invention: Bacillus Coagulans, High Cell Density Fermentation
Method thereof, and Method for Preparing Spray-dried Powder thereof
Technical Field [0001] The present invention relates to the field of microbial technology, in particular to a Bacillus coagulans, a high cell density fermentation method thereof, and the prepared spray-dried powder.
Description of the Related Art [0002] Microbial ecological agent is a microbial bacterial preparation made by special processes, and contains microorganisms beneficial to the host. The microbial ecological agent has the effects of being natural and non-toxic and promoting host health, and is widely applied to the fields of human healthcare and animal health.
[0003] Lactobacillus and bacillus microbial ecological agents are two major varieties which are most widely used in the feed industry. Lactobacillus species mainly includes lactic acid bacteria, bifidobacteria and etc., while bacillus species mainly includes Bacillus subtilis, Bacillus licheniformis and etc.
[0004] The lactobacillus microbial ecological agent plays roles by producing lactic acid, changing pH values in the intestinal tract, and conditioning intestinal microenvironment. This type of microbial ecological agent has poor stress resistance and is easy to lose activity during processing. The bacillus microbial ecological agent is capable of producing spores and tolerating unfavorable environment, such as high temperature, stomach acid and choline, and has strong stress resistance, but doesn’t possess the intestinal conditioning function of the lactobacillus.
[0005] Bacillus Coagulans, is a Gram-positive bacterium, and a lactic-acid-producing Bacillus. The characteristics of the Bacillus Coagulans are: 1. Bacillus Coagulans can produce lactic acid under an anaerobic environment of the intestinal tract to form an acidic environment and inhibit the growth of a pathogenic microorganism, which is beneficial to host health; 2. Bacillus Coagulans can exist in a spore state, tolerate high temperature, stomach acid and choline, and has strong environmental stress resistance. It can be said that Bacillus Coagulans has both the advantages of lactobacillus and bacillus microbial ecological agents.
[0006] Due to its good feeding effect and processing performance, Bacillus Coagulans has been included in a directory of feed additives, and becomes an important member of the feed microbial ecological agent industry.
[0007] More and more attentions have been paid to Bacillus Coagulans, and a great deal of researches have been conducted. ZL200710122111 .X discloses preparation of Bacillus Coagulans feed additives, and the viable count in the fermentation liquid reaches up to 5.0 x 109 cfu/mL. ZL 200810235512.0 discloses a liquid state fermentation process for Bacillus Coagulans CGMCC No.2602, in a 1T fermentor, the total bacteria count in the fermentation liquid is 6.0 X 109 cfu/mL, the spore count is 5.8 X109 cfu/mL, and the spore rate is 96.7%. ZL 201110428027.7 discloses a solid state fermentation process for Bacillus Coagulans CICC No.20138, the number of cells is 1.16X 101° cfu/g and the spore rate is 77.2%. ZL 201210464404.7 discloses a feedback feed-batch culture process for Bacillus Coagulans CGMCC No.6681, and the viable count in the fermentation liquid reaches up to 5.8 X109 cfu/mL.
[0008] Although a great deal of researches have been conducted in the industry, the selling price of Bacillus Coagulans in the market remains at a high level, which makes the product difficult to be popularized. This is because the industrial fermentation level of Bacillus Coagulans is low, cell loss during processing is high and the cell content in the finished product is low. These unfavorable factors directly result in a high product price and influence popularity and application of Bacillus Coagulans.
Technical Problems [0009] In view of the above problems in the prior art, the applicant provides a Bacillus coagulans, a high cell density fermentation method thereof, and a method for preparing a spray-dried powder. The newly-developed Bacillus coagulans according to the present invention has the characteristics of high cell density fermentation and high cell content, therefore, it is beneficial to reducing the product cost, is convenient for market popularization of products, and has a positive significance in promoting healthy livestock and poultry cultivation.
Solution
Technical Solution [0010] The technical solution of the present invention is as follows:
[0011] A Bacillus coagulans (Bacillus coagulans) provided by the present invention has been deposited at the China Center for Type Culture Collection with a deposit number of CCTCC M 2015273. Particularly, the present invention is implemented in the following steps:
[0012] 1. Screening of Bacillus coagulans [0013] 2g of soil sample near a dairy farm in Mashan, Wuxi, Jiangsu is weighed. 98 mL of sterile physiological saline is added. After heat treatment for 10 min at the temperature of 80°C, the mixture is oscillated on a shaker at a temperature of 37°C and at a speed of 120 rpm for 20 min, 1 mL of the supernatant is pipetted into an LB culture medium for dilution and coating, so as to isolate bacterial strains. The genus of the bacterial species is effectively and accurately determined through such means for the strain as morphological observation, physiological & biochemical experiments, and 16S rDNA analysis.
[0014] Finally, a strain with a serial number of ZS-007 is obtained, and identified as a Bacillus coagulans.
[0015] ZS-007 is deposited in the China Center for Type Culture Collection on May 3, 2015 with a deposit number of CCTCC M 2015273.
[0016] 2. Testing of Bacillus coagulans lactic-acid-producing capability [0017] Bacillus coagulans ZS-007 is inoculated into an acid-producing capability test culture medium, and cultured under static conditions at a temperature of 37 °C to conduct an acid-producing capability test. Compositions of the acid-producing capability test culture medium: 20 g/L glucose, 10 g/L peptone, 5 g/L yeast powder, 2 g/L potassium dihydrogen phosphate, 1 g/L magnesium sulfate and 0.5 g/L manganese sulfate. 10% NaOH is added to the culture medium to adjust pH to 7.0, and then the culture medium is sterilized at a temperature of 121 °C for 20 min.
[0018] The lactic acid content in the fermentation liquid is determined by HPLC, and the results are shown in FIG. 1. It can be seen from FIG. 1 that the lactic acid content is 10 g/L or so.
[0019] 3. High cell density fermentation of the Bacillus coagulans [0020] 1) Strain activation [0021] The Bacillus coagulans strain is inoculated into a nutrient agar slant culture medium, and incubated at a temperature of 25-37 °C for 24-48 hr; then streaked on a nutrient agar eggplant bottle slant, and incubated at a temperature of 25-37 °C for 24-48 hr.
[0022] Compositions of the nutrient agar slant culture medium and the nutrient agar eggplant bottle are both: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, 15-20g/L agar, with a pH of 7.2-7.4.
[0023] 2) Shake-flask culture [0024] The activated strain is inoculated into a shake-flask, and incubated with shaking for 18-30 hr.
[0025] Formulation of shake-flask culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4. 20 mL of liquid volume in 250mL shake-flask, and 50 mL of liquid volume in 500mL shake-flask.
[0026] Culture conditions: the shake-flask is rotated at a speed of 200 r/min and a temperature of 25-37 °C.
[0027] 3) Seed tank enlarged culture [0028] The strain after enlarged culture is used for later fermentation. Compositions of seed tank culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4; the seed tank is 10-1000L in volume, with 50-80% of liquid volume.
[0029] The culture is conducted at a stirring speed of 180-250 r/min, a ventilation amount of 0.5-1.0 vvm, and a temperature of 25-37 °C for 18-30hr.
[0030] 4) Fermentor fermentation [0031] The seed liquid with an inoculum size of 0.1%-10% is inoculated into the fermentor, the fermentation culture medium is composed of 25 g/L glucose, 15 g/L peptone, 10 g/L yeast powder, 1 g/L potassium dihydrogen phosphate, and 0.5 g/L magnesium sulfate, with an initial pH of 7.0. The fermentor is 1-50m3 in volume, with 50-80% of liquid volume.
[0032] The culture is conducted at a stirring speed of 100-200 r/min, a ventilation amount of 0.5-0.8 vvm, and a temperature of 25-37 °C for 16-48hr.
[0033] The fermentation liquid is taken for microscopic examination, when more than 90% of bacterial bodies form spores, i.e., a fermentation end-point is arrived, the final fermentation liquid is obtained.
[0034] The viable count and the spore count in the final fermentation liquid are determined with a spread plate method, and the spore rate is calculated. In particular, the spore rate is the ratio of the spore count and the viable count. The culture medium used is a nutrient agar culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, 15-20g/L agar, with a pH of 7.2-7.4. The viable count in the final fermentation liquid is determined after gradient dilution; after the final fermentation liquid is taken for heat treatment for 10 min at the temperature of 80°C and then gradient dilution, the spore count is determined. The viable count in the final fermentation liquid is 1.5*1Ο10 cfu/mL or above, the spore count is 1.4x1O10 cfu/mL or above, and the spore rate is 90.8% or above.
[0035] 4. Preparation of spray-dried powder [0036] Wet bacterial sludge is collected after the final fermentation liquid is centrifuged, and then spray-dried to obtain the dry bacterial powder. Operating conditions are: an air inlet temperature of 210 °C and an air outlet temperature of 100 °C. The viable count in the spray-dried powder is 8.0x1011 cfu/g or above.
[0037] The obtained dry bacterial powder can be mixed with starch, glucose or any other filler, such as a carrier conforming to standards to obtain a Bacillus coagulans microbial ecological agent. Beneficial Effects of the Present Invention.
Beneficial Effects [0038] The beneficial technical effects of the prevent invention lie in that:
[0039] By adopting the liquid fermentation process, the Bacillus coagulans viable count in the fermentation liquid is 1.52*101° cfu/mL or above, the spore count is 1.38*101° cfu/mL or above, and the spore rate is 90.8% or above. In the dry bacterial powder obtained by spray drying, the viable count is 8.0*1011 cfu/g or above. The spray-dried powder can be mixed with starch, glucose or any other filler to obtain a Bacillus coagulans microbial ecological agent.
[0040] By adopting the present invention, the production process is simple and stable, the viable count in the fermentation liquid is high, so that application cost can be directly reduced, and the product can be easily popularized, and the present invention has a significance in promoting a wide application of the Bacillus coagulans microbial ecological agent.
Brief Description of the Drawings Description of the Drawings [0041] FIG. 1 shows a lactic acid content of Bacillus coagulans in a fermentation liquid by an HPLC determination according to the present invention.
Embodiment
Embodiment of the Present Invention [0042] Embodiment 1: Screening of Bacillus coagulans [0043] 2g of soil sample near a dairy farm in Mashan, Wuxi, Jiangsu is weighed. 98 mL of sterile physiological saline is added. After heat treatment for 10 min at the temperature of 80°C, the mixture is oscillated on a shaker at a temperature of 37°C and at a speed of 120 rpm for 20 min, 1 mL of the supernatant is pipetted into an LB culture medium for dilution and coating, so as to isolate bacterial strains. The genus of the bacterial species is effectively and accurately determined through such means for the strain as morphological observation, physiological & biochemical experiments, and 16S rDNA analysis.
[0044] Finally, a strain with a serial number of ZS-007 is obtained, and identified as a Bacillus coagulans.
[0045] ZS-007 is deposited in the China Center for Type Culture Collection on May 3, 2015 with a deposit number of CCTCC M 2015273.
[0046] Embodiment 2: 1 m3 fermentor preparation [0047] Strain activation: the strain obtained in Embodiment 1 is inoculated into a nutrient agar slant culture medium, and incubated at a temperature of 30 °C for 24 hr; then streaked on a nutrient agar eggplant bottle slant, and incubated at a temperature of 30 °C for 24 hr. Compositions of the nutrient agar slant culture medium and the nutrient agar eggplant bottle are both: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, 15-20g/L agar, with a pH of 7.2-7.4.
[0048] Shake-flask culture: the activated strain is inoculated into a shake-flask, and incubated with shaking for 20 hr. Formulation of shake-flask culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4. 50 mL of liquid volume in 500mL conical flask. Culture conditions: the shake-flask is rotated at a speed of 200 r/min and a temperature of 30 °C.
[0049] Fermentor fermentation: the liquid in twenty conical flasks is collected as seed for inoculation in 1 m3 fermentor. The liquid volume is 65% of the 1 m3 fermentor. Compositions of culture medium: 25 g/L glucose, 15 g/L peptone, 10 g/L yeast powder, 1 g/L potassium dihydrogen phosphate, and 0.5 g/L magnesium sulfate, with an initial pH of 7.0. The cultured seed is inoculated into the fermentor for fermentation. The culture is conducted at a stirring speed of 200 r/min, a ventilation amount of 0.8 vvm, and a temperature of 30 °C for 40-44 hr.
[0050] The viable count in the fermentation liquid is 1.52*101° cfu/g, the spore count is 1.38*101° cfu/g, and the spore rate is 90.8 %.
[0051] Embodiment 3: 3 m3 fermentor preparation [0052] Strain activation is the same as that in Embodiment 2.
[0053] Shake-flask culture: the activated strain is inoculated into a shake-flask, and incubated with shaking for 20 hr. Formulation of shake-flask culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4. 20 mL of liquid volume in 250mL conical flask. Culture conditions: the shake-flask is rotated at a speed of 200 r/min and a temperature of 30 °C.
[0054] Seed tank enlarged culture: the seed liquid in ten 250mL shake-flasks is collected for seed tank inoculation. Compositions of seed tank culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4. The seed tank is 30 L in volume, with 80% of liquid volume. The culture is conducted at a stirring speed of 200 r/min, a ventilation amount of 0.8 vvm, and a temperature of 30 °C for 20 hr.
[0055] Fermentor fermentation: 3 m3 fermentor, with 70% of liquid volume. Compositions of the culture medium: 25 g/L glucose, 15 g/L peptone, 10 g/L yeast powder, 1 g/L potassium dihydrogen phosphate, and 0.5 g/L magnesium sulfate, with an initial pH of 7.0. The cultured seed is inoculated into the fermentor for fermentation. The culture is conducted at a stirring speed of 150 r/min, a ventilation amount of 0.7 vvm, and a temperature of 35 °C for 36-42 hr.
[0056] The viable count in the fermentation liquid is 1.79x101° cfu/g, the spore count is 1.72x101° cfu/g, and the spore rate is 96.1 %.
[0057] Embodiment 4: 30 m3 fermentor and preparation of spray-dried powder [0058] Strain activation is the same as that in Embodiment 2.
[0059] Shake-flask culture: the activated strain is inoculated into a shake-flask, and incubated with shaking for 20 hr. Formulation of shake-flask culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4. 50 mL of liquid volume in 500mL conical flask. Culture conditions: the shake-flask is rotated at a speed of 200 r/min and a temperature of 30 °C.
[0060] Seed tank enlarged culture: the seed liquid in twenty 500mL shake-flasks is collected for seed tank inoculation. Compositions of seed tank culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4. The seed tank is 500 L in volume, with 75% of liquid volume. The culture is conducted at a stirring speed of 200 r/min, a ventilation amount of 0.8 vvm, and a temperature of 30 °C for 20 hr.
[0061] Fermentor fermentation: 30 m3 fermentor, with 70% of liquid volume. Compositions of the culture medium: 25 g/L glucose, 15 g/L peptone, 10 g/L yeast powder, 1 g/L potassium dihydrogen phosphate, and 0.5 g/L magnesium sulfate, with an initial pH of 7.0. The cultured seed is inoculated into the fermentor for fermentation. The culture is conducted at a stirring speed of 100 r/min, a ventilation amount of 0.6 vvm, and a temperature of 35 °C for 40-45 hr.
[0062] The viable count in the fermentation liquid is 1.99x101° cfu/g, the spore count is 1.92x101° cfu/g, and the spore rate is 96.5 %.
[0063] Wet bacterial sludge is collected by centrifugation, and then spray-dried to obtain the dry bacterial powder. Operating conditions are: an air inlet temperature of 210 °C and an air outlet temperature of 100 °C. The viable count in the spray-dried powder is 8.32x1011 cfu/g.
[0064] Embodiment 5: Preparation of spray-dried powder [0065] The fermentation liquid of the Bacillus coagulans is obtained with the fermentation method in Embodiment 4, wet bacterial sludge is collected by centrifugation, and then spray-dried to obtain the dry bacterial powder. Operating conditions are: an air inlet temperature of 210 °C and an air outlet temperature of 100 °C. The viable count in the spray-dried powder is 6.54x1011 cfu/g.
[0066] Those skilled in the art can make various modifications and changes to the above description, and all of these modifications and changes belong to the protection scope defined by the appended claims of the present invention.

Claims (3)

[Claim 1] [Claim 2] [Claim 3] Claims
1) strain activation the Bacillus coagulans strain is inoculated into a nutrient agar slant culture medium, and incubated at a temperature of 25-37 °C for 24-48 hr; then streaked on a nutrient agar eggplant bottle slant, and incubated at a temperature of 25-37 °C for 24-48 hr; compositions of the nutrient agar slant culture medium and the nutrient agar eggplant bottle are both: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, 15-20g/L agar, with a pH of 7.2-7.4;
1. A Bacillus coagulans strain ZS-007, deposited at the China Center for Type Culture Collection with a deposit number of CCTCC M 2015273.
2) shake-flask culture the strain obtained in step 1) is inoculated into a shake-flask, and incubated with shaking for 18-30 hr; formulation of shake-flask culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4; 20 mL of liquid volume in 250mL shake-flask, and 50 mL of liquid volume in 500mL shake-flask; culture conditions: the shake-flask is rotated at a speed of 200 r/min and a temperature of 25-37 °C;
3) seed tank enlarged culture compositions of seed tank culture medium: 10 g/L peptone, 3g/L beef extract, 5 g/L sodium chloride, with a pH of 7.2-7.4; the seed tank is 10-1000L in volume, with 50-80% of liquid volume; the culture is conducted at a stirring speed of 180-250 r/min, a ventilation amount of 0.5-1.0 vvm, and a temperature of 25-37 °C for 18-30hr;
4) fermentor fermentation the seed liquid with an inoculum size of 0.1 %-10% obtained in step 3) is inoculated into the fermentor, the fermentation culture medium is composed of 25 g/L glucose, 15 g/L peptone, 10 g/L yeast powder, 1 g/L potassium dihydrogen phosphate, and 0.5 g/L magnesium sulfate, with an initial pH of 7.0; the fermentor is 1-50m3 in volume, with 50-80% of liquid volume; the culture is conducted at a stirring speed of 100-200 r/min, a ventilation amount of 0.5-0.8 vvm, and a temperature of 25-37 °C for 16-48hr; the fermentation liquid is taken for microscopic examination, when more than 90% of cells form spores, i.e., a fermentation end-point is arrived, the fermentation liquid is obtained.
2. A high cell density fermentation method of the Bacillus coagulans according to claim 1, characterized in that, the method particularly comprises the following steps:
3. A method for preparing a spray-dried powder based on the high cell density fermentation method of the Bacillus coagulans according to claim 2, characterized in that, wet bacterial sludge is collected after the fermentation liquid obtained in step 4) is centrifuged, and then spray-dried to obtain the dry bacterial powder; conditions of spray-drying are: an air inlet temperature of 210 °C and an air outlet temperature of 100 °C; the viable count in the spray-dried powder is 8.0x1011 cfu/g or above.
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CN104974966B (en) * 2015-07-22 2017-12-12 江南大学 A kind of bacillus coagulans and its fermentation process in high density and dry bacterium powder preparation method
CN105441354B (en) * 2015-10-28 2019-07-02 南京工业大学 A kind of preparation method of high activity bacillus coagulans active bacteria formulation
CN106801013A (en) * 2015-11-26 2017-06-06 瑞普(天津)生物药业有限公司 One kind mixing probiotics dried object and preparation method
CN108179117B (en) * 2016-12-09 2020-12-29 天津农学院 Method for producing brown mushroom by high-density fermentation
CN109207406A (en) * 2018-10-24 2019-01-15 广东绿百多生物开发有限公司 A kind of method and culture medium for improving bacillus coagulans cell and forming gemma
CN109593680A (en) * 2018-12-26 2019-04-09 武汉科诺生物科技股份有限公司 A kind of Dipel liquid fermentation medium and its bacterium powder and oil suspending agent
CN110117561B (en) * 2019-05-05 2020-07-28 中铁十八局集团有限公司 Compound biological agent for increasing survival rate of new trees and preparation method
CN110129230B (en) * 2019-05-20 2020-11-17 江南大学 Microbial preparation and application thereof in domestic sewage treatment of expressway service area

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001083704A1 (en) * 2000-05-02 2001-11-08 Biofermin Pharmaceutical Co., Ltd. Spray-dried microbial cells
WO2014081395A1 (en) * 2012-11-23 2014-05-30 Agency For Science, Technology And Research Highly Efficient Production Of Lactic Acid From Hemicellulose Sugars By Newly Isolated Thermophilic Bacillus Coagulans Strains

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101412983B (en) * 2008-11-20 2011-01-26 江苏省苏微微生物研究有限公司 Bacillus coagulans, preparation of high-density cultivated spore preparation, and use thereof
CN102505003B (en) * 2011-12-19 2013-06-26 湖南省微生物研究所 High-density fermenting method of bacillus coagulans for livestock and poultry, as well as preparation prepared by method and application thereof
CN102943057B (en) * 2012-11-16 2014-06-25 南京工业大学 Bacillus coagulans and process for high-density fermentation of bacillus coagulans
CN104232525A (en) * 2014-08-29 2014-12-24 湖北省生物农药工程研究中心 Process for preparing viable bacillus coagulans preparation
CN104974966B (en) * 2015-07-22 2017-12-12 江南大学 A kind of bacillus coagulans and its fermentation process in high density and dry bacterium powder preparation method

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001083704A1 (en) * 2000-05-02 2001-11-08 Biofermin Pharmaceutical Co., Ltd. Spray-dried microbial cells
WO2014081395A1 (en) * 2012-11-23 2014-05-30 Agency For Science, Technology And Research Highly Efficient Production Of Lactic Acid From Hemicellulose Sugars By Newly Isolated Thermophilic Bacillus Coagulans Strains

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