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Method for producing L-lactic acid by fermenting bagasse cellulose

A technology of cellulose and cellulase, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of environmental pollution, waste of renewable resources, etc., achieve stable acid production rate and good industrial application prospects , The effect of easy access to raw materials

Inactive Publication Date: 2014-03-12
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This not only wastes abundant renewable resources, but also pollutes the environment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The enzymatic hydrolysis of embodiment 1 bagasse cellulose

[0027] Pulverize bagasse with a pulverizer and pass through a 20-mesh sieve; accurately weigh 30 g of bagasse after sieving, and add 0.7% H 2 o 2 Mix solution with 4% NaOH, stir at 85°C for 4h; filter with suction, wash with distilled water until the filtrate is neutral, dry the residue in a drying oven at 105°C to constant weight, then pulverize with a pulverizer, pass through a 40-mesh sieve, Gather spares.

[0028] Accurately weigh 1 g of pretreated bagasse, add 7 mL of cellulase solution with a concentration of 10 mg / mL, substrate concentration of 40 g / L, and 18 mL of citric acid buffer solution with a pH of 5.0. Enzymolyzed in a water bath for 36 hours, then boiled to inactivate the enzyme, centrifuged, and the enzymolyzed solution was recovered for later use.

[0029] The concentration of reducing sugar in the enzymolysis solution was measured by DNS method, and the result showed that the concentratio...

Embodiment 2

[0030] Embodiment 2 free cell fermentation

[0031] Preparation of slant medium: Peel the potatoes, weigh 200g, add 1000mL of water, boil for 30min, filter, remove the potato pieces, make up the filtrate to 1000mL, add 20g of glucose, 20g of agar, heat to dissolve and divide into test tubes. Sterilize at 121°C for 20 minutes, place it at an angle, and cool it down;

[0032] Slant culture of strains: put the strain Rhizopus oryzae (Gim3.129) into a sterilized test tube PDA medium, cultivate it at 28°C for 5 days, and then store it in a refrigerator at 4°C;

[0033] Preparation of spore suspension: take a cultured slant strain, wash the spores on the slant with sterile normal saline, add sterile glass beads to a sterilized Erlenmeyer flask to fully disperse , filter with sterilized absorbent cotton, wash the filter residue with sterile water, make the volume of the filtrate 30mL, dilute until the spore suspension contains 1×10 7 spores / mL;

[0034] According to the following...

Embodiment 3

[0039] Embodiment 3 immobilized cell fermentation

[0040] Preparation of slant medium: Peel the potatoes, weigh 200g, add 1000mL of water, boil for 30min, filter, filter out the potato pieces, make up the filtrate to 1000mL, add 20g of glucose, 15g of agar, heat to dissolve and divide into test tubes. Sterilize at 121°C for 20 minutes, place it at an angle, and cool it down;

[0041] Slant culture of strains: put the strain Rhizopus oryzae (Gim3.129) into a sterilized test tube PDA medium, cultivate it at 28°C for 5 days, and then store it in a refrigerator at 4°C;

[0042] Preparation of spore suspension: take a cultured slant strain, wash the spores on the slant with sterile normal saline, add sterile glass beads to a sterilized Erlenmeyer flask to fully disperse , filter with sterilized absorbent cotton, wash the filter residue with sterile water, make the volume of the filtrate 30mL, dilute until the spore suspension contains 1×10 7 spores / mL;

[0043] Preparation ...

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Abstract

The invention discloses a method for producing L-lactic acid by fermenting bagasse cellulose. The method is characterized by using bagasse as a raw material, carrying out raw material pretreatment and cellulose enzymolysis and producing L-lactic acid by using moulds as fermentation strains and enzymatic hydrolysate of bagasse cellulose as a carbon source through fermentation with free cells or immobilized cells. The method has the beneficial effects that the raw material is accessible and is low in cost; the amount of reducing sugar obtained through enzymolysis is higher than that of reducing sugar in other reports, the fermentation period is short, the fermentation liquor is easy to separate, and the conversion rate of the product is high; not only can free fermentation be adopted but also immobilized fermentation can be carried out, and the immobilized cells for immobilized fermentation can be reused and have stable acid yields and high lactic acid conversion rates. In short, the method is suitable for industrially producing L-lactic acid by efficiently converting bagasse cellulose, greatly reduces the production cost of lactic acid, solves the problems of difficulty in control of a lactobacillus fermentation process and difficulty in separation and purification caused by the free cells, and has good industrial application prospects.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation to produce organic acids, in particular to a method for producing L-lactic acid by fermentation of bagasse cellulose. Background technique [0002] L-lactic acid (L-lactic acid) is the L-lactic acid converted from pyruvate through the catalysis of L-lactate dehydrogenase during microbial fermentation. Compared with D-lactic acid, because L-lactic acid can be directly absorbed by the human body without any side effects, it is widely used in food, medicine, chemical industry and other fields. [0003] At present, the microorganisms that ferment and produce L-lactic acid are mainly lactic acid bacteria and rhizopus. Lactic acid bacteria have high requirements for nutrients, and are facultative anaerobic microorganisms. It is difficult to control the fermentation process, and the optical purity of the produced L-lactic acid is not high. Traditional fermentation mostly adopts the method...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/56C12R1/845
Inventor 孙卫东唐艳琼戴莉唐湘毅苏芬芬
Owner GUANGXI UNIV
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