title: High-Efficiency Secretion of β-Mannanase in Bacillus subtilis through Protein Synthesis and Secretion Optimization.
abstract: The manno endo-1,4-mannosidase (β-mannanase, EC. 3.2.1.78) catalyzes the random hydrolysis of internal (1 → 4)-β-mannosidic linkages in the mannan polymers. A codon optimized β-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 β-mannanase was observed using SPlipAas a signal peptide. Then a 1.25-fold activity of β-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 Pmglv, a modified maltose-inducible promoter, significantly elevated the production of β-mannanase. After 72 h of flask fermentation, the enzyme activity of β-mannanase in the supernatant when using locust bean gum as substrate reached 2207 U/mL. This work provided a promising β-mannanase production strain in industrial application.
authors.affiliation: Tianjin Institute of Industrial Biotechnology and ‡Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences , Tianjin 300308, P. R. China.|Department of Chemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen , 9713 AV, Groningen, The Netherlands.
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Song
authors.affiliation: Tianjin Institute of Industrial Biotechnology and ‡Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences , Tianjin 300308, P. R. China.
authors.forename: Gang
authors.initials: G
authors.lastname: Fu
authors.affiliation: Tianjin Institute of Industrial Biotechnology and ‡Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences , Tianjin 300308, P. R. China.
authors.forename: Huina
authors.initials: H
authors.lastname: Dong
authors.affiliation: National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology , Beijing 100190, China.|Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences , Beijing 100190, China.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Li
authors.affiliation: National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology , Beijing 100190, China.|Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences , Beijing 100190, China.
authors.forename: Yuguang
authors.initials: Y
authors.lastname: Du
authors.affiliation: Tianjin Institute of Industrial Biotechnology and ‡Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences , Tianjin 300308, P. R. China.
authors.forename: Dawei
authors.identifier: 0000-0002-5857-394X
authors.initials: D
authors.lastname: Zhang
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D021381:Protein Transport; D044902:beta-Mannosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D044902:beta-Mannosidase
keywords: Bacillus subtilis; chaperones; promoters; signal peptidase; signal peptides; β-mannanase