Method for synchronous enzymolysis and fermentation production of D-lactic acid by using cottonseed protein
The technology of cottonseed protein and neutral protease is applied in the field of fermentation and production of D-lactic acid to achieve the effect of simplifying operation steps and reducing the cost of raw materials
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Embodiment 1
[0047] Embodiment 1, the determination of the optimum neutral protease concentration of efficient production D-lactic acid
[0048] The components of the fermentation medium used in this example are as follows: 55g / L glucose, 40g / L cottonseed protein, 33g / L calcium carbonate, 0g / L, 0.1g / L, 0.2g / L, 0.3g / L, 0.4g / L Or 0.5g / L neutral protease, the solvent of the culture medium is water; pH5.5-6.5. Each concentration is the final concentration of the corresponding component in the fermentation medium. The fermentation medium needs to be sterilized by autoclaving, specifically at 115° C. for 20 minutes.
[0049] The specific steps to determine the optimum concentration of neutral protease for the production of D-lactic acid are as follows:
[0050] 1) Seed fungus culture: Sporolactobacillus laelacticus DSM442 stored on MRS solid medium was put into a test tube containing 6mL seed medium with an inoculation loop, cultured at 30°C for 48h, and then measured by volume fraction 10% o...
Embodiment 2
[0055] Example 2, Determination of the optimum cottonseed protein concentration for efficient production of D-lactic acid
[0056] The components of the fermentation medium used in this embodiment are as follows: 68g / L glucose, 40g / L calcium carbonate, 0.2g / L (equivalent to 1×10 4 U / L) neutral protease, 10g / L, 20g / L, 30g / L, 40g / L, 50g / L or 60g / L cottonseed protein, the medium solvent is water; pH5.5-6.5. Each concentration is the final concentration of the corresponding component in the fermentation medium. The fermentation medium needs to be sterilized by autoclaving, specifically at 115° C. for 20 minutes.
[0057] The specific steps to determine the optimum cottonseed protein concentration for the production of D-lactic acid are as follows:
[0058] 1) Seed fungus cultivation: the method is the same as step 1) in Example 1.
[0059] 2) The seed solution in step 1) was added to the above six fermentation media (50 mL) containing different concentrations of cottonseed prot...
Embodiment 3
[0063] Example 3, Determination of the optimum temperature for efficient production of D-lactic acid
[0064] The components of the fermentation medium used in this example are as follows: 75g / L glucose, 40g / L cottonseed protein, 45g / L calcium carbonate, 0.2g / L (equivalent to 1×10 4 U / L) neutral protease, the solvent is water; pH5.5-6.5. Each concentration is the final concentration of the corresponding component in the fermentation medium. The fermentation medium needs to be sterilized by autoclaving, specifically at 115° C. for 20 minutes.
[0065] The temperature settings for the fermentation culture were as follows: 30°C, 37°C and 42°C.
[0066]The specific steps to determine the optimum temperature for producing D-lactic acid are as follows:
[0067] 1) Seed fungus cultivation: the method is the same as step 1) in Example 1.
[0068] 2) Add the seed liquid from step 1) to the above-mentioned fermentation medium (50mL) at a volume ratio of 1:10. Place them in Erlenmey...
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