Fermentation broth processing method capable of improving final concentration of pullulanase

A pullulanase and treatment method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems that the enzyme concentration cannot be further increased, the viscosity of the pullulanase fermentation broth is high, etc., and achieve improvement Filtration performance, reduction of storage space, effect of increasing concentration

Active Publication Date: 2020-01-10
JIANGNAN UNIV
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Problems solved by technology

The viscosity of the pullulanase fermentation broth prepared by the recombinant bacteria Bacillus amyloliquefaciens / pUC980-BdP is relatively high. After the final enzyme liquid is concentrated to 1 / 5 of the original volume, if the ultrafiltration operation is continued, the volume of the fermentation liquid cannot be further reduced, and the enzyme concentration cannot be further increased

Method used

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  • Fermentation broth processing method capable of improving final concentration of pullulanase

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Embodiment 1

[0021] Embodiment 1: the influence of different processing methods of pullulanase fermented liquid on ultrafiltration concentration

[0022] Take the supernatant of the pullulanase fermentation broth produced by fermentation in a 5 L fermenter, and make four portions of 900 mL each, labeled A1, A2, B, and C, respectively. Add 100 mL of double distilled water to A1 and A2, add 100 mL of 10× citric acid buffer solution to B, and add 100 mL of 10× fermentation broth treatment solution C to C.

[0023] The 10X citric acid buffer added in B contained 200 mM citric acid-sodium citrate, pH 5.4-5.6. The configuration method is: add 800mL deionized water to a 1 L volumetric flask, add citric acid monohydrate ((C 6 h 8 o 7 )·H 2 (2) 12.6 g, sodium citrate monohydrate ((Na 3 C 6 h 5 o 7 )·H 2 (2) 41.2g, if the pH deviates then adjust the pH to 5.5±0.1 with NaOH or HCl, add water to 1 L.

[0024] After mixing, the four mixed solutions are processed as follows:

[0025] A1 was p...

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Abstract

The invention discloses a fermentation broth processing method capable of improving final concentration of pullulanase, and belongs to the technical field of enzyme engineering and fermentation engineering. According to the fermentation broth processing method capable of improving final concentration of pullulanase, thermal insulation is carried out after mixing of a treatment solution C with a pullulanase fermentation liquid supernatant; and then, ultra-filtration concentration is carried out so as to obtain pullulanase fermentation broth with high enzyme activity. Being applied in post-extraction stage of pullulanase prepared by performing fermentation with recombinant bacillus amylolyticus BF7658/pUC980-BdP, the fermentation broth processing method is capable of improving ultrafiltration performance of the pullulanase fermentation broth. Being subjected to ultra-filtration concentration, the enzyme activity of the concentrate prepared from the fermentation broth liquid supernatant by the fermentation broth processing method disclosed by the invention is higher than 1.75 times of the enzyme activities of concentrates prepared from fermentation broths which are not yet treated with the treatment solution C after the same ultra-filtration concentration process.

Description

technical field [0001] The invention relates to a fermentation liquid treatment method for increasing the final concentration of pullulanase, and belongs to the technical fields of enzyme engineering and fermentation engineering. Background technique [0002] Pullulanase (Pullulanase, EC3.2.1.41) has the function of hydrolyzing α-1,6 glycosidic bonds in starch, and in the production of glucose, the synergistic effect with glucoamylase can increase the yield of glucose. Recombinant Bacillus amyloliquefaciens / pUC980- BdP [Sun Juanjuan, Shen Weishi, Guiyang Wang Zhengxiang. Secretion and expression of pullulanase gene in Bacillus amyloliquefaciens BF7658 strain. Industrial Microbiology, 2011, 41(6): 18-22], which has a good ability to produce pullulanase . [0003] In the production of industrial enzyme preparations, in order to obtain products with high enzyme concentration, it is often necessary to concentrate the fermentation broth by ultrafiltration to remove part of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/44C12N1/20C12R1/07
CPCC12N9/2457C12Y302/01041C12N1/20
Inventor 祝冬君德青美朵王施岚孙娟娟沈微陈献忠杨海泉夏媛媛
Owner JIANGNAN UNIV
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