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Adaptive evolution method for gluconobacter oxydans efficiently utilizing non-glucose carbon sources and application of adaptive evolution method

A technology of Gluconobacter oxydans and adaptive evolution, applied in the fields of biotechnology and microorganisms, can solve the problems of affecting the production efficiency of sugar acid, low utilization efficiency, etc.

Active Publication Date: 2020-05-12
SHANXI SYNTHETIC BIOLOGY INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the glucose effect, the utilization efficiency of non-glucose carbon sources by Gluconobacter oxydans is low, which greatly affects the overall sugar and acid production efficiency

Method used

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  • Adaptive evolution method for gluconobacter oxydans efficiently utilizing non-glucose carbon sources and application of adaptive evolution method
  • Adaptive evolution method for gluconobacter oxydans efficiently utilizing non-glucose carbon sources and application of adaptive evolution method
  • Adaptive evolution method for gluconobacter oxydans efficiently utilizing non-glucose carbon sources and application of adaptive evolution method

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Experimental program
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Effect test

Embodiment 1

[0016] Example 1 strain screening

[0017] Under sterile conditions, the parent strain of Gluconobacter oxidans was inoculated into the detoxified corn stalk hydrolyzate, and cultured at 30°C and 220rpm for 24 hours. The fermentation pH changed from a certain amount of CaCO 3 maintenance; then insert 10% inoculum into the non-detoxified corn stalk hydrolyzate and culture for 24 hours under the same conditions, and then alternately transfer in the detoxified and non-detoxified hydrolyzate until the fermentation performance of the strain is significantly improved and maintained Stablize. The obtained strain has been named Gluconobacter oxydans RM7, and was deposited in the General Microorganism Center (CGMCC) of the China Committee for the Collection of Microbial Cultures (CGMCC) on October 11, 2017, with the preservation number CGMCC No.14801.

Embodiment 2

[0018] The utilization of embodiment 2 bacterial strain to single carbon source

[0019] Adaptive evolution of new strains to the utilization of a single carbon source was investigated in a synthetic medium. The initial sugar concentration is 40g / L, the nutrient salt component is 10.0g / L yeast extract, 1.5g / L KH 2 PO 4 ,1.5g / L(NH 4 ) 2 SO 4 and 0.5g / LMgSO 4 7H 2 O. The culture condition is 50mL system cultured at 200rpm and 30°C for 72h. The analysis results showed that the single carbon source utilization efficiency of the adaptive evolutionary strains was improved compared with the original strains.

Embodiment 3

[0020] Example 3 Bacterial Strain Utilization of Total Sugar from Hydrolyzate

[0021] The domesticated strains were also tested for fermentation in hydrolyzate. Fermentation was carried out in a 5L reactor, the fermentation conditions were 30% solid content, 4mg enzyme protein / g dry material, 500rpm, 35°C, aeration rate 1vvm and pH 4.8. The sugar-acid fermentation is directly carried out under the condition of fully saccharified and hydrolyzed mash without solid-liquid separation. The analysis results showed that similar to the single carbon source synthetic medium fermentation, the utilization of all non-glucose carbon sources in the hydrolyzate was significantly improved, especially mannose, which is difficult to utilize in the hydrolyzate, and the original The strain needs to be cultured at a high density to achieve complete conversion of mannose, while the adaptive evolution strain can convert most of the mannose within 72 hours of fermentation.

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Abstract

The invention discloses an adaptive evolution method for gluconobacter oxydans efficiently utilizing non-glucose carbon sources and application of the adaptive evolution method, and belongs to the biotechnology and microbial technology field. The method comprises the concrete steps that firstly, the gluconobacter oxydans is cultured in a hydrolysate containing inhibitor lignocellulose, then the cultured gluconobacter oxydans is transferred into a hydrolysate without the inhibitor for further culture, and the gluconobacter oxydans is alternately transferred in such a way until the fermentationperformance of a strain reaches stable. The new strain obtained by acclimatization has obviously improved utilization efficiency of the non-glucose carbon sources, and the strain is named as gluconobacter oxydans RM7, and the collection number of the strain is CGMCC No.14801; and the method is simple and efficient, and has important referential significance in breeding of strains containing an inhibitor lignocellulose system.

Description

【Technical field】 [0001] The invention relates to the fields of biotechnology and microorganism technology, and relates to a method for the adaptive evolution of Gluconobacter oxydans that efficiently utilizes non-glucose carbon sources and its application. 【Background technique】 [0002] Lignocellulosic raw materials are composed of cellulose, hemicellulose and lignin. Among them, cellulose can be hydrolyzed into glucose by cellulase, and hemicellulose can be degraded into xylose, mannose and galactose by acid pretreatment or under the action of hemicellulase. The hemicellulose-derived sugars account for about 40% of the total sugars. Common strains can only use glucose and cause a large loss of other fermentable sugars. These unused sugars also increase the COD content in the process wastewater, resulting in a more severe sewage treatment load. [0003] Gluconobacter oxydans has a unique redox ability to convert various sugars into corresponding sugar acids. Its membran...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/36C12N1/20C12R1/01
CPCC12N1/36C12N1/20
Inventor 鲍杰侯伟亮姚瑞苗张宏森金慈张建
Owner SHANXI SYNTHETIC BIOLOGY INST CO LTD
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