High-Efficiency Secretion of beta-Mannanase in Bacillus subtilis through Protein Synthesis and Secretion Optimization

Yafeng Song, Gang Fu, Huina Dong, Jianjun Li, Yuguang Du, Dawei Zhang

Research output: Contribution to journalArticleAcademicpeer-review

41 Citations (Scopus)

Abstract

The manno endo-1,4-mannosidase (beta-mannanase, EC. 3.2.1.78) catalyzes the random hydrolysis of internal (1 -> 4)-beta-mannosidic linkages in the mannan polymers. A codon optimized beta-mannanase gene from Bacillus licheniformis DSM13 was expressed in Bacillus subtilis. When four Sec-dependent and two Tat-dependent signal peptide sequences cloned from B. subtilis were placed upstream of the target gene, the highest activity of P-mannanase was observed using SPlipA,A as a signal "peptide. Then a 1.25-fold activity beta-mannanase was obtained when another copy of groESL operon was inserted into the genome of host strain. Finally, five different promoters were separately used to enhance the synthesis of the target protein. The results showed that promoter P-mglv a modified maltose-inducible promoter, significantly elevated the production beta-mannanase. After 72 h of flask fermentation, the enzyme activity of P-mannanase in the supernatant when using locust bean gum as substrate reached 2207 U/mL. This work provided a promising beta-mannanase production strain in industrial application.

Original languageEnglish
Pages (from-to)2540-2548
Number of pages9
JournalJournal of Agricultural and Food Chemistry
Volume65
Issue number12
DOIs
Publication statusPublished - 29-Mar-2017

Keywords

  • beta-mannanase
  • Bacillus subtilis
  • signal peptides
  • promoters
  • chaperones
  • signal peptidase
  • METHYL PARATHION HYDROLASE
  • GRAM-POSITIVE BACTERIA
  • EXTRACELLULAR PRODUCTION
  • ESCHERICHIA-COLI
  • ALPHA-GALACTOSIDASE
  • EXPRESSION SYSTEMS
  • SIGNAL PEPTIDES
  • VECTORS
  • TRANSFORMATION
  • LICHENIFORMIS

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