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Method for producing lactic acid and simultaneously realizing coupling preparation of GABA (gamma-aminobutyric acid)

A technology of lactic acid and sodium glutamate, applied in the field of biochemical engineering, can solve the problems of solidification, consumption of industrial raw materials, difficult to continue fermentation, etc., and achieve the effects of removing product inhibition, reducing production costs, and overcoming a large amount of calcium sulfate waste residue and waste water.

Active Publication Date: 2014-05-07
SHANDONG YANGCHENG BIOLOGY TECH CO LTD
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, when using CaCO 3 When neutralizing the lactic acid produced in lactic acid fermentation, a large amount of CaCO in the fermentor 3 It is easy to block the feeding port; in the middle and late stages of fermentation, the high concentration of calcium lactate makes the fermentation liquid viscous, generating a large amount of foam and causing liquid escape; or solidification occurs, making it difficult to continue the fermentation
In addition, using CaCO 3 As a neutralizer, there are problems such as long extraction process, high consumption of industrial raw materials and low recovery rate, and at the same time, a large amount of calcium sulfate waste residue and waste water are generated, environmental protection pressure is high, processing costs are increased, and energy consumption is high.

Method used

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  • Method for producing lactic acid and simultaneously realizing coupling preparation of GABA (gamma-aminobutyric acid)

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Preparation of immobilized glutamic acid decarboxylase engineering bacteria:

[0037] (1) Pick the engineered bacteria of glutamate decarboxylase (E.coli BL21(DE3) / pET28a-gadB, from the activated LB solid medium, (see "Cloning, sequencing and expression of a glutamate decarboxylase gene from the GABA-producing strain Lactobacillus brevis CGMCC1306[J].ANNALS OF MICROBIOLOGY.2012,62(2):689-698") monoclonal cells were injected with kanamycin (50μg·mL -1 ) in LB liquid medium at 37°C with 200r / min shaking overnight to obtain seed liquid.

[0038] (2) Inoculate the seed liquid with 1% inoculum volume fraction into the -1 ) in LB medium, cultured at 37°C, 200r / min until OD 600 Add IPTG at 0.6 o'clock to make the final concentration of IPTG 0.5 μM, induce culture at 30° C., 150 r / min for 6 h.

[0039] (3) Treat the bacterial cells expressing glutamic acid decarboxylase at 70°C for 30 minutes, then collect the bacterial cells by centrifugation to obtain permeabilized glutami...

Embodiment 2

[0042] (1) Inoculate 2% (v / v) Rhizopus oryzae As3.819 spore suspension (5×10 6 cells / mL) in the seed medium, 30°C, 220r / min shaking on the shaker for 24 hours, to obtain the seed liquid.

[0043] (2) Inoculate the seed liquid into 50mL fermentation medium with 4% inoculation amount, and cultivate it at 32°C and 200r / min. When the pH of the fermentation liquid is lower than 4.8, add immobilized corn to the fermentation liquid. Amino acid decarboxylase engineering bacteria (dry cell weight content of bacteria is 23.2mg, total activity of glutamic acid decarboxylase is 150U), pyridoxal phosphate with a final concentration of 0.1mM, and L-sodium glutamate is continuously added at the same time, so that The pH value of the fermentation broth was maintained between 4.8-5.0 throughout the whole process, and the cultivation was continued for 60 hours. At the same time, the fermentation broth without immobilized glutamic acid decarboxylase was used as blank control.

[0044] The cont...

Embodiment 3

[0046] (1) Inoculate 2% (v / v) Rhizopus oryzae spore suspension (5×10 6 cells / mL) in the seed medium, 30°C, 220r / min shaking on the shaker for 24 hours, to obtain the seed liquid.

[0047] (2) Inoculate the seed liquid into 50mL fermentation medium with 4% inoculation amount, and cultivate it at 32°C and 200r / min. When the pH of the fermentation liquid is lower than 4.8, add immobilized corn to the fermentation liquid. Amino acid decarboxylase thallus (the dry cell weight content of the bacterium is 23.2mg, the total activity of glutamic acid decarboxylase is 150U), the final concentration is 0.1mM pyridoxal phosphate, and L-sodium glutamate is continuously added at the same time, so that The pH value of the fermentation broth was maintained between 5.2-5.6 throughout the whole process, and the cultivation was continued for 60 hours. At the same time, the fermentation broth without immobilized glutamic acid decarboxylase was used as blank control.

[0048] The measured conten...

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Abstract

The invention discloses a method for producing lactic acid and simultaneously realizing coupling preparation of GABA (gamma-aminobutyric acid). The method comprises the steps of inoculating rhizopus oryzae into a fermentation culture medium to perform fermentation, adding glutamic acid decarboxylase, pyridoxal phosphate and L-sodium glutamate into a fermentation solution when the pH value of the fermentation solution is less than 4.8, then continuously adding L-sodium glutamate into the fermentation solution, and generating GABA of glutamic acid decarboxylase by utilizing the vitality of catalyzing glutamic acid to consume H+ in an environment so as to maintain the pH value of the fermentation solution at 4.8-6.0 and achieve the purposes of relieving acid inhibition against fermentation of lactic acid and realizing simultaneous production of lactic acid and GABA. The invention provides a new method for relieving the inhibition of a substrate for fermentation of lactic acid; furthermore, in the fermentation process, the coupling production of a functional compound, namely gamma-aminobutyric acid, is realized, so that the yield of lactic acid is increased, and the economy of the process is further improved.

Description

technical field [0001] The invention belongs to the field of biochemical engineering, in particular to a method for producing lactic acid and simultaneously preparing GABA. Background technique [0002] Lactic acid is an important industrial raw material, which has been widely used in food, medicine, cosmetics, agriculture and chemical industry for a long time. In addition, lactic acid can be used to prepare engineering materials such as polylactic acid. And polylactic acid can be used to make completely biodegradable plastics, produce green packaging materials and agricultural films, in order to eliminate the environmental crisis caused by "white pollution". In addition, due to its good biocompatibility, polylactic acid is also widely used in the field of pharmaceutical industry, including medical sutures, drug sustained release materials, bone fixation repair materials and other tissue engineering materials. As the market demand for polylactic acid continues to increase,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/56C12P13/00
Inventor 梅乐和赵伟睿黄俊胡升雷引林胡桂香金志华
Owner SHANDONG YANGCHENG BIOLOGY TECH CO LTD
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