A kind of bacillus subtilis glycerol kinase mutant gene glpk and its application
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A Bacillus subtilis, mutation gene technology, applied in the field of engineering bacteria, to achieve the effects of clear genetic background, improved ability, and increased riboflavin accumulation level
Active Publication Date: 2021-04-09
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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[0004] The present invention proposes a Bacillus subtilis glycerol kinase mutant gene glP And application thereof, have solved the technical problem that Bacillus subtilis utilizes glycerol in the prior art
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[0034] Step 1: Screen the starting strains of the Bacillus subtilis system without antibiotic markers B. subtili s 168 Δupp and the construction of basic operation plasmid pSS
[0035] Bacillus subtilis starting strain used in the present invention B. subtili s 168 Δupp from B. subtilis 168. The basic operation plasmid pSS is derived from pUC18. For the detailed construction process of the two, please refer to Shi, T., Wang, G., Wang, Z., Fu, J., Chen, T., Zhao, X., 2013. Establishment of a Markerless Mutation Delivery System in Bacillus subtilis Stimulated by a Double-Strand Break in the Chromosome. PLoS one. 8, e81370.
[0036] The present invention does not limit the construction method of the basic operation plasmid pSS and the selection of the resistance gene.
[0037] Step 2: Engineering Strains B. subtilis M270I build process
[0038] (1) to the strain B. subtili s 168 Δupp Introduce genetic mutations without trace on the genome glP (G810T) The ope...
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Abstract
The invention relates to the field of engineering bacteria, in particular to a glycerol kinase mutant gene of Bacillus subtilis glP and its application. The mutant gene glP The base sequence is shown in SEQ ID NO:1. The present invention contains the glycerol kinase mutant gene of Bacillus subtilis glP (G810T) engineered bacteria are biologically safe, with clear genetic background and contained mutant genes glP (G810T) can greatly increase the glycerol utilization level of Bacillus subtilis, and can increase the specific growth rate of the strain by 11% and the maximum biomass by more than 16% under the condition of basic glycerol salt shake flask culture.
Description
technical field [0001] The invention relates to the field of engineering bacteria, in particular to a glycerol kinase mutant gene glpK of Bacillus subtilis and its application. Background technique [0002] Glycerol molecules contain a higher carbon content (52.8%), and high reducing properties (reducing power is twice that of glucose), which can produce a higher yield of reducing compounds, so it can be used as a cheap alternative to the microbial fermentation industry carbon source. Microorganisms ferment glycerol to produce different chemicals such as 1,3-propanediol, dihydroxyacetone phosphate, 1,2-propanediol, propionic acid, 2,3-butanediol, lactic acid, formic acid, acetic acid, ethanol, butyric acid, Chemicals such as butanol, succinic acid, pyruvate, phosphoenolpyruvate, etc. Genetically engineered bacteria with bacteria as host bacteria have the advantages of short fermentation cycle, simple raw material requirements, and mature genetic engineering technology. In...
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