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Method for improving fermentation property of malic acid

A malic acid and seed technology, applied in the field of microorganisms, can solve the problems of high cost and unsuitable for industrial production, and achieve the effects of improving fermentation performance, being beneficial to dissolved oxygen and mass transfer, and increasing the quantity

Active Publication Date: 2018-09-28
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct fermentation method is also called the one-step fermentation method. This process uses sugary raw materials and molds to directly ferment malic acid to produce malic acid; the two-step fermentation method uses two microorganisms with different functions to use sugary raw materials. Mold fermentation produces fumaric acid or a mixture of fumaric acid and malic acid, which is then converted into a single L-malic acid by yeast or bacteria; conversion fermentation is the conversion of fumaric acid, the first fermentation product of the two-step fermentation, into apples acid, and the enzymatic conversion method uses microbial fumarase to convert fumarate into malate, which is costly and not suitable for industrial production

Method used

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  • Method for improving fermentation property of malic acid
  • Method for improving fermentation property of malic acid
  • Method for improving fermentation property of malic acid

Examples

Experimental program
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Effect test

Embodiment 1

[0035] (1) Preliminary isolation and purification of Aspergillus oryzae

[0036]Soil samples were collected from orchards in the surrounding areas of Wuxi, an appropriate amount of soil was taken, put into a 250mL triangular flask filled with 100mL sterile water, shaken, and diluted 10 -4 ~10 -6 were respectively spread on the acid production indicator plate and incubated at 28°C for 3 days. Observe the discoloration of the plate, pick a single colony spore with a large discoloration circle and inoculate it on a solid screening medium, culture it at 28°C for 5 days, separate and purify it until there are no bacteria, and store it on a PDA slant for use.

[0037] (2) Screening of L-malate-producing Aspergillus oryzae

[0038] Take 10 spore-filled bacterial strains obtained from the above isolation and purification, add 5 mL of sterile water to each, scrape off all the spores with an inoculation loop in an ultra-clean bench, shake them up with glass beads, and filter through r...

Embodiment 2

[0042] (1) Slant culture: Inoculate a ring of strains on the slant medium and culture at 28°C for 5 days;

[0043] (2) Seed culture: insert the spore suspension into the seed culture medium with a filling volume of 100mL / 250mL, so that the final concentration of spores is 10 6 cells / mL; cultured for 14 hours at a temperature of 30°C and a shaker speed of 200r / min;

[0044] (3) Fermentation culture: the seed solution was inoculated into the fermentation medium at 10% (v / v), and fermented for 120 h at a temperature of 30° C. and a shaker speed of 200 r / min.

[0045] Fermentation medium (g / L): glucose 100, peptone 6, K 2 HPO 4 0.15, KH 2 PO 4 0.15, MgSO 4 ·7H 2 O0.1, CaCl 2 2H 2 O 0.1, CaCO 3 100.

[0046] The contents of L-malic acid and heteroacids in the fermentation broth were detected by HPLC, and the results are shown in Table 1.

[0047] Table 1 L-malic acid and impurity content in the fermentation broth

[0048]

[0049] Adopt the method of present embodime...

Embodiment 3

[0051] Fermentation medium (g / L): glucose 100, peptone 6, K 2 HPO 4 0.15, KH 2 PO 4 0.15, MgSO 4 7H 2 O 0.1, CaCl 2 2H 2 O 0.1, CaCO 3 90.

[0052] Incline culture, seed culture and fermentation culture conditions are the same as in Example 2.

[0053] Add alumina 0.3g / L to the fermentation medium.

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Abstract

The invention discloses a method for improving the fermentation property of malic acid, belonging to the field of microorganism. An aspergillus oryzae strain FMME 338 is assigned with the accession number of CCTCC NO:M 2016401. The invention also discloses a method for fermentation production of L-malic acid, the temperature is 30 DEG C, the rotation speed of a shaking table is 200r / min, the fermentation period is 120h, finally, the yield of the L-malic acid through fermentation can achieve 129.2g / L, and the strain belongs to an excellent wild strain with high yield of L-malic acid. The methodfor fermentation production of L-malic acid is simple and feasible in technological operation, the cost of the culture medium is low, meanwhile, the purity of L-malic acid in the fermentation liquidis high, the amount of heteroacid is little, the fermentation period is short, and the method is suitable for industrial production.

Description

technical field [0001] The invention relates to a method for improving malic acid fermentation performance, belonging to the field of microorganisms. Background technique [0002] Malic acid, also known as dihydroxysuccinic acid, has the molecular formula HOOCH 2 -CH(OH)COOH, the relative molecular mass is 134.09. Malic acid is one of the very important organic acids, widely used in chemical, food and pharmaceutical industries and other fields. Because its molecular structure contains an asymmetric carbon atom, it has optical activity, and there are two chiral enantiomers of L-malic acid and D-malic acid. Among them, L-malic acid widely exists in organisms and participates in cell metabolism as a member of the tricarboxylic acid cycle. [0003] Malic acid is an important part of natural fruit acids. Malic acid has obvious taste effect, long-lasting and soft sour taste, and stable property. It is an excellent sour agent and acid regulator. L-malic acid can improve the st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/46C12R1/69
CPCC12P7/46C12N1/145C12R2001/69
Inventor 刘立明丁强陈修来周洁罗秋玲刘佳
Owner JIANGNAN UNIV
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