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- W2351725457 abstract "The manA1 gene encoding mature β-mannanase and the manA2 gene contained a signal peptide from the Bacillus subtilis JNA 3-10 were amplified.The two genes were inserted into expression vector pMA5,and the plasmid pMA5-manA1 and pMA5-manA2 were constructed and transformed into B.subtilis 168 respectively.The recombinant strains BPM1001(pMA5-manA1 / B.subtilis 168) and BPM1002(pMA5-manA2/B.subtilis 168) were therefore obtained.SDS-PAGE showed that the genes manA1 and manA2 were expressed successfully in recombinant B.subtilis 168.After incubated in shake-flask,the enzyme activity of strain BPM1002 was 9.65-fold higher than that of BPM1001.Subsequently,the β-mannanase gene manA3 was cloned with His-Tag added downstream of the gene manA2.Then the enzyme was purified successively by Ni affinity chromatography.The analysis of enzymatic properties showed that the optimum activity of the β-mannanase was at pH 6.5 and 65 ℃,and the enzyme activity was maintained 77.8% of original activity after incubated at 37 ℃ for 30 days.The activity of the β-mannanase reached 2 748.82 U/mL in a 5 L fermentor.Fig 9,Tab 3,Ref 19" @default.
- W2351725457 created "2016-06-24" @default.
- W2351725457 creator A5074999356 @default.
- W2351725457 date "2012-01-01" @default.
- W2351725457 modified "2023-09-25" @default.
- W2351725457 title "Cloning and Expression of β-mannanase Gene in Bacillus subtilis" @default.
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- W2351725457 hasPublicationYear "2012" @default.
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