CN101792727A - Bacillus coagulans and application thereof in L-sodium lactate preparation - Google Patents
Bacillus coagulans and application thereof in L-sodium lactate preparation Download PDFInfo
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Abstract
The invention discloses a bacillus coagulans, and the name of the bacillus coagulans is bacillus coagulans CASN, was preserved in a China Center for Type Culture Collection, and the preservation registration number is CCTCC M 2010035. The invention also discloses the application of the bacillus coagulans in L-sodium lactate preparation, i.e. L- sodium lactate is prepared by culturing and fermenting the bacillus coagulans. The highest concentration of the L- sodium lactate prepared from the bacillus coagulans of the invention can reach 162g/l, and the highest volume production efficiency can reach 5.2g/l/h.
Description
Technical field
The present invention relates to a kind of special bacterium of the L-of preparation Sodium.alpha.-hydroxypropionate and the application of described bacterium thereof, relate in particular to a bacillus coagulans and the application in the L-sodium lactate preparation thereof; Belong to the food fermentation production technical field.
Background technology
Sodium.alpha.-hydroxypropionate, molecular formula C
3H
5NaO
3, molecular weight is 112.06, purified Sodium.alpha.-hydroxypropionate is colourless or intimate colourless clarified liq.Sodium.alpha.-hydroxypropionate is widely used in industries such as food, medicine and makeup.In food service industry, Sodium.alpha.-hydroxypropionate can be used as food preservative, emulsifying agent, wetting Agent for Printing Inks, flavour improvers, quality improver, anti-oxidant synergistic agent and pH regulator agent, can strengthen growth, the prolongation product shelf-lives of malignant bacteria in local flavor, the inhibition food, part substitutes Sodium Benzoate and makes aseptic applications in food service industry abroad; In pharmaceutical industries, Sodium.alpha.-hydroxypropionate is mainly as injection, and its major function is for replenishing body fluid and mediator's cylinder electrolyte balance; At cosmetic industry, the L-Sodium.alpha.-hydroxypropionate is widely used as the moisturizing agent of skin care product, can make skin keep moisture, reduces wrinkle.A unsymmetrical carbon is arranged in the Sodium.alpha.-hydroxypropionate molecule, therefore have opticity.The L-Sodium.alpha.-hydroxypropionate is a levorotation, and the D-Sodium.alpha.-hydroxypropionate is a dextrorotatory, and the DL-Sodium.alpha.-hydroxypropionate is a racemism.Have only the L-Sodium.alpha.-hydroxypropionate to be utilized by human body, the production technology of therefore developing L-sodium lactate with high optical purity has great importance.
Sodium.alpha.-hydroxypropionate also can be used for synthesizing lactic acid, and then produces degradable polymer---poly(lactic acid).Poly(lactic acid) has biocompatibility, biodegradability and good mechanical property and physicals, thereby thought the new packaging material that the new millennium is the most rising by industrial community, to be expected to progressively substitute materials such as polyethylene, polypropylene, polystyrene, application prospect is very wide.
Find through retrieval prior art, Chinese patent literature CN1076725C, day for announcing 1993-09-29, put down in writing a kind of " breeding of aerobic organism or the method for aerobic fermentation ", this technology utilizes yellow soda ash or sodium hydroxide and lactic acid direct reaction to carry out the technology that Sodium.alpha.-hydroxypropionate is produced, and the prerequisite of this method is that highly purified lactic acid will be arranged, traditional DL-lactic acid low price, but the L-lactic acid of high-optical-purity costs an arm and a leg, and therefore is not suitable for the production with the L-Sodium.alpha.-hydroxypropionate; Biological process mainly is to utilize the microorganism of lactic acid producing to ferment, and adding lime carbonate or calcium hydroxide carry out neutralization reaction in the process of fermentation, obtain calcium lactate, and then generates Sodium.alpha.-hydroxypropionate with the yellow soda ash reaction.Also having a kind of method is to utilize sodium hydroxide or yellow soda ash to be neutralizing agent in the process of fermentation, and what obtain in the fermented liquid like this is Sodium.alpha.-hydroxypropionate directly just, but owing to the toxic action of Sodium.alpha.-hydroxypropionate to thalline, the output of fermentation is not high usually.In addition, the configuration of the Sodium.alpha.-hydroxypropionate of biological process production depends on employed microorganism, and the bacterial strain that can obtain L-sodium lactate with high optical purity at present is actually rare, and Bacillus coagulans is a kind of bacterial strain that produces high-optical-purity L-lactic acid.
Further retrieval is found, Chinese patent literature CN101173242, open day 2008-5-7, put down in writing a kind of " producing the method and the coagulate bacillus cereus special thereof of L-lactic acid ", this technology utilizes a bacillus coagulans (Bacillus coagulans CASHCGMCC № 2184) can produce L-lactic acid efficiently, but utilize lime carbonate to be neutralizing agent in this patent, actual in the fermented liquid what obtain is calcium lactate.
U.S. Patent number 5079164 and Chinese patent literature CN1498265A have put down in writing two kinds of methods of utilizing Bacillus coagulans Bacillus coagulans DSM 5196 and Bacillus coagulans Bacillus coagulans SIM-7DSM 14043 to carry out lactic acid-producing respectively, but the product that this technology finally obtains also is a calcium lactate.
Summary of the invention
The present invention is directed to the prior art above shortcomings, the object of the invention provides a bacillus coagulans and the application in the L-sodium lactate preparation thereof.
Bacillus coagulans of the present invention laboratory called after Bacillus coagulans (Bacillus coagulans) CASN, this bacterial strain screen the soil near perfumery, Shanghai International Automobile City Tourist Festival district and mutagenesis obtains.Described Bacillus coagulans (Bacilluscoagulans) CASN has been preserved in Chinese typical culture collection center, and (be called for short CCTCC, the address is: wuchang, wuhan Luo Jiashan), preservation registration number was CCTCC M 2010035 on 01 29th, 2010.
The application of Bacillus coagulans of the present invention in the L sodium lactate preparation; Be by described Bacillus coagulans being cultivated and the back acquisition L-Sodium.alpha.-hydroxypropionate that ferments.
Wherein:
Described cultivation is meant a kind of enforcement in the following dual mode:
A) slant culture that carries out successively and seed culture;
B) directly carry out seed culture.
Described slant culture is meant: described Bacillus coagulans is inoculated on the solid slant culture base of the agar that contains 1.5～2.0g/100ml 45～60 ℃ of culture temperature, incubation time 8～12 hours;
Preferably 50～55 ℃ of described yeast culture temperature;
Preferably 9～10 hours described yeast culture time;
Described seed culture is meant: the product or the described Bacillus coagulans that obtain with slant culture are inoculated in the seed culture medium, 45～60 ℃ of culture temperature, and training method is cultivated 120 rev/mins of rotating speeds, incubation time 6～8 hours for utilizing the Scroll-tupe shaking table;
Contain in every liter of the described seed culture medium: glucose 60～80 grams, yeast powder 10～15 grams, soy peptone 5～8 grams, lime carbonate 10～20 grams, surplus is a water; The pH of described seed culture medium is 5.5～6.
Preferably 50～55 ℃ of described yeast culture temperature;
Preferably 6～7 hours described yeast culture time;
Described fermentation is meant: make seed liquor with described Bacillus coagulans, then seed liquor is inserted fermention medium, with 45～60 ℃ of temperature, and bubbling air 0.1～1.5vvm cultivated 12～24 hours, stop ventilation afterwards, continue to cultivate 18～48 hours, the control mixing speed must contain the fermentation culture of L Sodium.alpha.-hydroxypropionate at 100～500 rev/mins in the fermenting process.Further preferred mode is: make seed liquor with described Bacillus coagulans, then seed liquor is inserted fermention medium, with 50～55 ℃ of temperature, and bubbling air 0.1～0.5vvm cultivated 18～36 hours, stop ventilation afterwards, continue to cultivate 24～48 hours, the control mixing speed must contain the fermentation culture of L-Sodium.alpha.-hydroxypropionate at 200～300 rev/mins in the fermenting process.
Wherein, utilizing sodium hydroxide or yellow soda ash control pH in the described fermenting process is 5.5～6.0.
Contain glucose 80～170 grams per liters, nitrogenous source 5～20 grams per liters in the above-mentioned fermention medium, surplus is a water; Wherein said nitrogenous source is meant: a kind of or its any part by weight combination in yeast powder 5～15 grams per liters, soy peptone 5～15 grams per liters and cottonseed protein 10～20 grams per liters.
Experiment confirm: near Bacillus coagulans (Bacillus coagulans) the more existing Bacillus coagulans of CASN CCTCC M 2010035 pairs of L-Sodium.alpha.-hydroxypropionates tolerance (Bacillus coagulans) the CASH CGMCC № 2184 that the present invention screens the soil perfumery, Shanghai International Automobile City Tourist Festival district and mutagenesis obtains is significantly improved.Concrete, Bacillus coagulans (Bacilluscoagulans) CASN CCTCC M 2010035 and Bacillus coagulans (Bacillus coagulans) CASH CGMCC № 2184 cultivated 20 hours respectively in L-Sodium.alpha.-hydroxypropionate pressure substratum after, the dry weight of bacterial strain CASH is 0.77 grams per liter, and the dry weight of bacterial strain CASN reaches 1.3 grams per liters, improved 68.8% than bacterial strain CASH, comparative result is seen Fig. 1.
The present invention utilizes Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035, realized with glucose being substrate, under 45～60 ℃ of conditions with the L-Sodium.alpha.-hydroxypropionate of the efficient fermentative production optical purity of 90～95% glucose acid invert ratio more than 99%, test result shows: L-Sodium.alpha.-hydroxypropionate concentration reaches as high as 162 grams per liters, volume production is most effective reach 5.2 grams per liters/hour, ventilate and control pH value by changing during the fermentation in addition, can effectively improve the tolerance of bacterial strain to Sodium.alpha.-hydroxypropionate, and the generation of minimizing by product, improve transformation efficiency.
Description of drawings
Fig. 1 is the comparison synoptic diagram of 2184 pairs of L-Sodium.alpha.-hydroxypropionates of bacterial strain homocoagulation genus bacillus of the present invention (Bacillus coagulans) CASH CGMCC № tolerance.
Fig. 2 is the change curve of glucose in the preparation process of the present invention and L-Sodium.alpha.-hydroxypropionate.
Embodiment
Below embodiments of the invention are elaborated, present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
Separation mutagenesis screening and the evaluation of Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035
1, separates mutagenesis screening
Obtain soil sample near perfumery, Shanghai International Automobile City Tourist Festival district, per 1 gram soil sample adds in 50 milliliters of nutrient broth mediums (glucose 50 grams per liters, yeast powder 10 grams per liters, pH are 6), 50 ℃ of enrichment culture 8～12 hours.Dilution is applied in the culture dish that contains nutrient agar (lime carbonate 20 grams per liters, agar powder 10 grams per liters, pH are 6 for glucose 50 grams per liters, yeast powder 10 grams per liters) then, cultivates 24 hours for 50 ℃.After waiting to grow single bacterium colony, 280 of the big bacterium colonies of choosing colony area and transparent circle area, be inoculated into fermention medium (glucose 150 grams per liters respectively, yeast powder 20 grams per liters, lime carbonate 80 grams per liters, pH are 6) in, 50 ℃ of shaking tables were cultivated 48 hours, measure the output of L-lactic acid, select the highest bacterial strain of output, be used for next step ultraviolet mutagenesis.
In nutrient broth medium, 50 ℃ of shaking tables are cultured to mid-log phase with the inoculation tentatively selected, suspend furnishing 10 with physiological saline washing back
8Individual bacterium/milliliter, get 5 milliliters in 9 centimetres of culture dish, apart from ultraviolet lamp 30 centimeters difference mutagenesis 15 seconds, 30 seconds, 1 minute and 5 minutes, draw 100 microlitres respectively the mutagenesis bacterium liquid that under variant mutagenic condition, obtains, be applied on the nutrient agar, under the condition of dark, cultivated 14 hours for 50 ℃, with physiological saline the bacterium on the culture dish is washed, getting 100 microlitres respectively inserts in the nutrient broth medium (glucose 50 grams per liters, yeast powder 10 grams per liters, L-Sodium.alpha.-hydroxypropionate 50 grams per liters) that contains 5% (/ 100 milliliters of 5 grams) L-Sodium.alpha.-hydroxypropionate, 50 ℃ of shaking tables are cultivated, select one group of the fastest nutrient solution of growth, dilute this nutrient solution, be coated on nutritive solid substratum (glucose 50 grams per liters that contain 2% (/ 100 milliliters of 2 grams) L-Sodium.alpha.-hydroxypropionate, yeast powder 10 grams per liters, L-Sodium.alpha.-hydroxypropionate 20 grams per liters, agar 18 grams per liters) on the flat board, wait to grow single bacterium colony after, the bacterium colony that the choosing colony area is big, be inoculated in the nutrient broth medium that contains the 5%L-Sodium.alpha.-hydroxypropionate, 50 ℃ of shaking tables are cultivated, and select the fastest bacterial strain of growth.Finally obtain the strongest bacterial strain of L-Sodium.alpha.-hydroxypropionate tolerance.
2, the evaluation of bacterial strain
According to the authentication method that " Bai Jie system identification handbook (the 8th edition) " and " common bacteria system identification handbook " (Science Press) provides the strongest bacterial strain of above-mentioned L-Sodium.alpha.-hydroxypropionate tolerance is identified, qualification result shows that this bacterium is shaft-like, long 2.5～4 microns, wide 0.8～0.9 micron, the non-sphere of spore, the tooth sporangiocyst does not expand.Can under anaerobic grow, can under pH5.7, grow, can not can not grow at 5% sodium-chlor in the growth of 2% sodium-chlor 65 ℃ of growths 60 ℃ of growths.Glucose, pectinose, N.F,USP MANNITOL can be utilized, wood sugar, fructose can not be utilized, can hydrolyzed starch, polychrom, can not caseinhydrolysate, tween 80, gelatinum, the arginine hydrolase positive.The result of this bacterial strain fatty acid analysis is with typical Bacillus coagulans.
Based on above feature, the identification of strains that above-mentioned L-Sodium.alpha.-hydroxypropionate tolerance is the strongest is the lactobacillus that condenses (Bacilluscoagulans), laboratory name Bacillus coagulans (Bacillus coagulans) CASN, this bacterial strain has been preserved in Chinese typical culture collection center and (has been called for short CCTCC on 01 29th, 2010, the address is: wuchang, wuhan Luo Jiashan), preservation registration number is CCTCC M 2010035.
Embodiment 2
The comparison of 2184 pairs of L-Sodium.alpha.-hydroxypropionates of Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035 homocoagulation genus bacillus (Bacilluscoagulans) CASH CGMCC № tolerance
With Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035 and Bacillus coagulans (Bacilluscoagulans) CASH CGMCC № 2184 respectively at L-Sodium.alpha.-hydroxypropionate pressure substratum (glucose 50 grams per liters, yeast powder 10 grams per liters, L-Sodium.alpha.-hydroxypropionate 0.5 mol) cultivates in, after 20 hours, the dry weight of bacterial strain CASH is 0.77 grams per liter, and the dry weight of bacterial strain CASN reaches 1.3 grams per liters, has improved 68.8% than bacterial strain CASH, and concrete comparative result is seen Fig. 1.
Embodiment 3
Utilize Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035 to produce the L-Sodium.alpha.-hydroxypropionate 5 liters of fermentation cylinder for fermentation
Used substratum is composed as follows among this embodiment:
Slant medium: glucose 20 grams per liters, yeast extract 5 grams per liters, peptone 10 grams per liters, extractum carnis 10 grams per liters, bitter salt 0.58 grams per liter, four anhydrous manganeses, 0.25 grams per liter, lime carbonate 15 grams per liters, agar powder 15 grams per liters.The initial pH of this substratum is 6.5.Sterilized 20 minutes for 115 ℃.
Seed culture medium: glucose 70 grams per liters, yeast powder 10 grams per liters, soy peptone 5 grams per liters, lime carbonate 15 grams per liters.The initial pH of this substratum is 6.5.Sterilization is 20 minutes under 115 ℃ of conditions.
Fermention medium: glucose (the sugared company limited of Zibo rainbow space industry) 120 grams per liters, yeast powder (Beijing extensive and profound in meaning star biotechnology responsibility company limited, cat. no 01-013) 10 grams per liters, soy peptone (Beijing extensive and profound in meaning star biotechnology responsibility company limited, cat. no 01-004) 2 grams per liters.The initial pH of this fermention medium is 6.5, and is unsterilised.
The method of this fermentation production of L-lactic acid sodium may further comprise the steps:
(1) slant culture: with Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035 bacterial classification inoculations on slant medium, under 50 ℃ of conditions, static cultivation 10 hours;
(2) seed culture: with the bacterial strain of step (1) cultivation, encircle in the 300ml triangular flask that the 100ml seed culture medium is housed with inoculation articulating 2 under aseptic condition, under 50 ℃ of conditions, shaking table was cultivated 6 hours, 120 rev/mins of rotating speeds (13mm rotation radius) make seed liquor;
(3) fermentation culture: (BIOSTAT B B.Braun) adds 4 liters of fermention mediums in 5 liters of fermentor tanks to German Bei Lang.In fermention medium, insert the 400ml seed culture fluid, 200 rev/mins of fermentor tank mixing speed (33mm rotation radius), the feeding amount of air is 0.5vvm, stops after 16 hours ventilating, utilizing the pH of the sodium hydroxide control fermented liquid of 10M is 6.5,50 ℃ of condition bottom fermentations 30 hours.10 repetitions are established in experiment altogether.Got fermented liquid in per 3 hours, 6,000 rev/mins centrifugal 5 minutes, get supernatant liquor and detect L-Sodium.alpha.-hydroxypropionate concentration, D-Sodium.alpha.-hydroxypropionate concentration, glucose concn in the fermented liquid, calculate the optical purity of glucose acid invert ratio, volume production efficient and L-Sodium.alpha.-hydroxypropionate.
Wherein, the determination of glucose method is that fermented liquid dilution back is centrifugal, adopts bio-sensing analyser SBA-40C (Shandong Scientific Research Academy) to measure.
L-Sodium.alpha.-hydroxypropionate and D-Sodium.alpha.-hydroxypropionate Determination on content method are to adopt Agilent 1100 liquid chromatographs, outfit chiral separation post (separation of optics allosome is used for Mitsubishi chemical company, MCI GEL-CRS10W (3 μ) 4.6ID * 50mm).The concrete operations condition is: 0.002mol/L copper sulfate is as moving phase, flow 0.5ml/min, and sample size 20 μ L, UV-detector detects wavelength 254nm, 25 ℃ of service temperatures.Utilize L-Sodium.alpha.-hydroxypropionate and D-Sodium.alpha.-hydroxypropionate standard substance to make typical curve, calculate the content of L-Sodium.alpha.-hydroxypropionate and D-Sodium.alpha.-hydroxypropionate in the fermented liquid again according to typical curve.
Among the present invention, be the product of German Sigma-Aldrich company as the D-Sodium.alpha.-hydroxypropionate of standard substance, its article No. is 77166-1G; As the L-Sodium.alpha.-hydroxypropionate of standard substance is the product of German Sigma-Aldrich company, and its article No. is 71718-10G.Under as above chromatographic condition, D-lactic acid retention time is 10.150 minutes, and L-lactic acid retention time is 12.293 minutes.
Optical purity (optical purity) be weigh in the opticity sample enantiomorph surpass another enantiomorph amount measure its available enantiomeric excess value (enantiomeric excess) expression.The optical purity of L-lactic acid is calculated as follows among the present invention: (L-lactic acid content-D-lactic acid content) ÷ (L-lactic acid content+D-lactic acid content) * 100%.
Glucose acid invert ratio is defined as: the consumption (grams per liter) * 100% of L-lactic acid production (grams per liter) ÷ glucose.
The volume production definitions of efficiency is: L-Sodium.alpha.-hydroxypropionate output (grams per liter) ÷ fermentation time (hour).
After the fermentation ends, the concentration of glucose is 1.4 grams per liters ± 0.6 grams per liter, L-Sodium.alpha.-hydroxypropionate concentration 134 grams per liters ± 1.8 grams per liters, glucose acid invert ratio 91.0% ± 0.5%, volume production efficient 4.5 grams per liters/hour ± 0.4 grams per liter/hour, L-Sodium.alpha.-hydroxypropionate optical purity 99.5% ± 0.1%.
Embodiment 4
Utilize Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035 to produce the L-Sodium.alpha.-hydroxypropionate 5 liters of fermentation cylinder for fermentation
The composition of the substratum of used inclined-plane and seed and training method are with embodiment 3 among this embodiment.
Fermentation initial medium: glucose (the sugared company limited of Zibo rainbow space industry) 108 grams per liters, soy peptone (Beijing extensive and profound in meaning star biotechnology responsibility company limited, cat. no 01-004) 10 grams per liters, cottonseed protein (Beijing Kang Mingwei substratum technology limited liability company) 8 grams per liters.The initial pH of this fermention medium is 6.5, and is unsterilised.
Fermentation culture: (BIOSTAT B B.Braun) adds 4 liters of fermentation initial mediums in 5 liters of fermentor tanks to German Bei Lang.In fermention medium, insert the 400ml seed culture fluid, 300 rev/mins of fermentor tank mixing speed (33mm rotation radius), the feeding amount of air is 0.2vvm, stops after 12 hours ventilating, utilize the pH of the yellow soda ash control fermented liquid of 10M to ferment altogether 24 hours under 6.0,55 ℃ of conditions.Centrifugal 5 minutes of 6,000 rev/mins of fermented liquids are got supernatant liquor and are detected L-Sodium.alpha.-hydroxypropionate concentration, D-Sodium.alpha.-hydroxypropionate concentration, glucose concn in the fermented liquid, calculate glucose acid invert ratio, volume production efficient and L-Sodium.alpha.-hydroxypropionate optical purity.
After the result shows fermentation ends, the concentration of glucose is 1.1 grams per liters ± 0.2 grams per liter, L-Sodium.alpha.-hydroxypropionate concentration 125 grams per liters ± 2.2 grams per liters, glucose acid invert ratio 93.5% ± 0.4%, volume production efficient 5.2 grams per liters/hour ± 0.3 grams per liter/hour, L-Sodium.alpha.-hydroxypropionate optical purity 99.5% ± 0.2%.
Embodiment 5
Utilize Bacillus coagulans (Bacillus coagulans) CASN CCTCC M 2010035 to produce L-lactic acid 30 liters of fermentation cylinder for fermentation
The composition of the substratum of used inclined-plane and seed and training method are with embodiment 3 among this embodiment.
Fermentation initial medium: glucose (the sugared company limited of Zibo rainbow space industry) 160 grams per liters, yeast powder (Beijing extensive and profound in meaning star biotechnology responsibility company limited, cat. no 01-013) 5 grams per liters, cottonseed protein (Beijing Kang Mingwei substratum technology limited liability company) 20 grams per liters.The initial pH of this fermention medium is 6.5, and is unsterilised.
Fermentation culture: Shanghai is protected and is added 20 liters of fermentation initial mediums in emerging 30 liters of fermentor tanks.In fermention medium, insert 2 liters of seed culture fluids, 150 rev/mins of fermentor tank mixing speed (33mm rotation radius), the feeding amount of air is 1vvm, stops after 20 hours ventilating, utilize the pH of the yellow soda ash control fermented liquid of 10M to ferment altogether 48 hours under 6.2,52 ℃ of conditions.Wherein, ferment (after testing by 26 hours, the concentration of glucose is 19 grams per liters in the fermented liquid at this moment), stop to ventilate, add yeast powder, add cottonseed protein according to the amounts of adding 3 gram yeast powders in every liter of fermented liquid according to the amounts of adding 6 gram cottonseed proteins in every liter of fermented liquid; Be 60 gram yeast powders (Beijing extensive and profound in meaning star biotechnology responsibility company limited, cat. no 01-013) in each fermentor tank, 120 gram cottonseed proteins (Beijing Kang Mingwei substratum technology limited liability company).10 repetitions are established in experiment altogether.Centrifugal 5 minutes of 6,000 rev/mins of fermented liquids are got supernatant liquor and are detected L-Sodium.alpha.-hydroxypropionate concentration, D-Sodium.alpha.-hydroxypropionate concentration, glucose concn in the fermented liquid, calculate glucose acid invert ratio, volume production efficient and L-Sodium.alpha.-hydroxypropionate optical purity.
The change curve that the result shows glucose and L-Sodium.alpha.-hydroxypropionate in Bacillus coagulans (Bacillus coagulans) the CASN CCTCC M 2010035 fermentation production of L-lactic acid processes as shown in Figure 2.After the fermentation ends, the concentration of glucose is 15.0 grams per liters ± 2.8 grams per liters, L-Sodium.alpha.-hydroxypropionate concentration 162 grams per liters ± 5.2 grams per liters, glucose acid invert ratio 90.2% ± 1.3%, volume production efficient 3.4 grams per liters/hour ± 0.3 grams per liter/hour, L-lactic acid optical purity 99.7% ± 0.3%.
Claims (7)
1. a bacillus coagulans, this bacterium is called Bacillus coagulans (Bacillus coagulans) CASN, is preserved in Chinese typical culture collection center on 01 29th, 2010, and its preservation registration number is CCTCC M 2010035.
2. the application of the described Bacillus coagulans of claim 1 in the L-sodium lactate preparation.
3. application according to claim 2, it is characterized in that, make seed liquor with described Bacillus coagulans, then seed liquor is inserted fermention medium, with 45～60 ℃ of temperature, and bubbling air 0.1～1.5vvm cultivated 12～24 hours, stop ventilation afterwards, continue to cultivate 18～48 hours, the control mixing speed must contain the fermentation culture of L Sodium.alpha.-hydroxypropionate at 100～500 rev/mins in the fermenting process.
4. application according to claim 3, it is characterized in that, make seed liquor with described Bacillus coagulans, then seed liquor is inserted fermention medium, with 50～55 ℃ of temperature, and bubbling air 0.1～0.5vvm cultivated 18～36 hours, stop ventilation afterwards, continue to cultivate 24～48 hours, the control mixing speed must contain the fermentation culture of L-Sodium.alpha.-hydroxypropionate at 200～300 rev/mins in the fermenting process.
5. according to claim 3 or 4 described application, it is characterized in that utilizing sodium hydroxide or yellow soda ash control pH in the described fermenting process is 5.5～6.0.
6. according to claim 3 or 4 described application, it is characterized in that contain glucose 80～170 grams per liters, nitrogenous source 5～20 grams per liters in the described fermention medium, surplus is a water.
7. application according to claim 6 is characterized in that, described nitrogenous source is meant: a kind of or its any part by weight combination in yeast powder 5～15 grams per liters, soy peptone 5～15 grams per liters and cottonseed protein 10～20 grams per liters.
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