β-mannanase gene, recombinant expression vector, bacterial strain, β-mannanase and its preparation method and application
A technology of mannanase and expression vector, which is applied in the direction of microorganism-based methods, botany equipment and methods, biochemical equipment and methods, etc. It can solve the problems of low enzyme activity and poor high temperature resistance of β-mannanase , to achieve the effect of improving expression and broad application prospects
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[0025] The invention provides a β-mannanase gene, a recombinant expression vector, a bacterial strain, a β-mannanase and its preparation method and application, aiming at solving the problem of poor high temperature resistance of β-mannanase in the prior art, live low problem.
[0026] Construction of expression vector of single copy β-mannanase gene
[0027] 1.1 Synthesis of gene fragments of β-mannanase
[0028] According to the existing β-mannanase gene (derived from Thermotoga maritima), artificially synthesized β-mannanase gene fragment, its nucleotide sequence is shown in SEQ ID NO:1 . The amino acid sequence corresponding to the artificially synthesized β-mannanase is shown in SEQ ID NO:2.
[0029] The gene sequence design of the artificially synthesized β-mannanase of the present invention mainly considers the following points:
[0030] 1) By replacing the original low-frequency codons with high-frequency codons in Pichia pastoris (P.pastoris);
[0031] 2) reduced...
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