Recombinant bacillus subtilis strain expressing and secreting beta-mannanase efficiently and method for obtaining strain

A technology of Bacillus subtilis and mannanase, applied in the field of molecular biology, can solve the problem of predicting the most effective signal peptide sequence without downstream protein sequence, affecting the correct folding of β-mannanase, and β-mannanase Degradation and other issues
CN110004166AInactive Publication Date: 2019-07-12TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
Publication Date
2019-07-12
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the technical field of molecular biology, and in particular, relates to a recombinant bacillus subtilis strain expressing and secreting beta-mannanase efficiently and a methodfor obtaining the strain. Specifically, the method comprises the steps: a codon-optimized beta-mannanase gene is cloned to the downstreams of different signal peptide sequences in a pMA5 vector to secrete to extracellular sites; then restriction factors involved in the process of secretion are overexpressed in a host bacteria genome; and finally promoters on the expression vector are replaced, and the production of beta-mannanase is increased at the level of protein synthesis by enhancing transcription and translation processes. The recombinant bacillus subtilis expressing and secreting beta-mannanase efficiently utilizes a 2*SR culture medium for fermentation production of mannanase under conditions of the temperature of 37 DEG C and the rotating speed of 220 rpm, the highest enzyme activity of beta-mannanase in 72 h fermentation broth can reach 2207 U / mL, and the recombinant bacillus subtilis strain is a strain with good application prospect.
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Description

technical field

[0001] The invention belongs to the technical field of molecular biology, and specifically relates to one or more recombinant Bacillus subtilis strains highly expressing and secreting β-mannanase and an obtaining method thereof. Background technique

[0002] Bacillus subtilis (Bacillus subtilis) is a Gram-positive bacterium and an important non-pathogenic industrial microorganism. Its whole genome sequencing work has been completed, and its genetic background and physiological characteristics are clear. The use of Bacillus subtilis to express foreign proteins has many advantages: (1) high safety, and can be used in industrial production such as food and medicine; (2) can directly release secreted proteins into the medium, which is beneficial for separation and purification; (3) has The ability to efficiently secrete the target protein has a variety of secretory pathways, and the relevant secretory components in the pathway have been analyzed; the secreted re...

Claims

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