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Construction and characterization of a novel maltose inducible expression vector in Bacillus subtilis

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Abstract

A maltose-inducible expression vector in Bacillus subtilis has been developed and characterized. The vector permitted β-galactosidase expression at a high level (maximum activity, 8.16 U/ml) when induced and its expression was markedly repressed by glucose. Using this vector, we successfully expressed the other two genes, bioA and vgb. This thus provided a potential expression system for cloned genes in B. subtilis.

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References

  • Bolhuis A, Sorokin A, Azevedo V, Ehrlich SD, Braun PG, De Jong A, Venema G, Bron S, Van Dijl JM (1996) Bacillus subtilis can modulate its capacity and specificity for protein secretion through temporally controlled expression of sips gene for signal peptidase I. Mol Microbiol 22:605–618

    Article  PubMed  CAS  Google Scholar 

  • Bron S (1990) Plasmids. In: Harwood CR, Cutting SM (eds) Molecular biological methods for Bacillus. John Wiley and Sons, New York pp 75–174

    Google Scholar 

  • Bron S, Bolhuis A, Tjalsma H, Holsappel S, Venema G, van Dijl JM (1998) Protein secretion and possible role for multiple signal peptidases for precursor processing in Bacilli. J Biotechnol 64:3–13

    Article  PubMed  CAS  Google Scholar 

  • Dikshit KL, Yutaka O, Navani N, Patel S, Huang H, Stark BC, Webster DA (1998) Site-directed mutagenesis of bacterial hemoglobin: the role of glutamine (T7) in oxygen-binding in the distal heme pocket. Arch Biochem Biophys 349:161–166

    Article  PubMed  CAS  Google Scholar 

  • Eijnenburg CM, Bron S, Venema G (1987) Structural plasmid instability in recombination- and repair-deficient strains of Bacillus subtilis. Plasmid 17:167–170

    Article  Google Scholar 

  • Eima R, Venema G, Bron S (1996) A positive selection vector for the analysis of structural plasmid instability in Bacillus subtilis. Plasmid 35:14–30

    Article  Google Scholar 

  • Hartl B, Wehrl W, Wiegert T, Homuth G, Schumann W (2001) Development of a new integration site within the Bacillus subtilis chromosome and construction of compatible expression cassettes. J Bacteriol 183:2696–2699

    Article  PubMed  CAS  Google Scholar 

  • Hirata H, Fukazawa T, Negoro S, Okada H (1986) Structure of a β-galactosidase gene of Bacillus stearothermophilus. J. Bacteriol 166: 722–727

    PubMed  CAS  Google Scholar 

  • Kim L, Mogk A, Schumann W (1996) A xylose-inducible Bacillus subtilis integration vector and its application. Gene 87:53–61

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Schrogel O, Allmansberger R (1997) Optimisation of the BgaB reporter system: determination of transcriptional regulation of stress responsive genes in Bacillus subtilis. FEMS Microbiol Lett 153:237–243

    Article  PubMed  CAS  Google Scholar 

  • Shimotsu H, Henner DJ (1986) Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene 43:85–94

    Article  PubMed  CAS  Google Scholar 

  • Streit WR, Entcheva P (2003) Biotin in microbes, the genes involved in its biosynthesis, its biochemical role and perspectives for biotechnological production. Appl Microbiol Biotechnol 61:21–31

    PubMed  CAS  Google Scholar 

  • Thompson J, Pikis A, Ruvinov SB, Henrissat B, Yamamoto H, Sekiguchi J (1998) The gene glvA of Bacillus subtilis 168 encodes a metal-requiring NADH-dependent 6-phospho-glucosidase. J Biol Chem 273:27347–27356

    Article  PubMed  CAS  Google Scholar 

  • Tjalsma H, Bolhuis A, Jongbloed J, Bron S, Dijl J M (2000) Signal peptide-dependent protein transport in Bacillus subtilis: a genome-based survey of the secretome. Microbiol Mol Biol Rev 64:515–547

    Article  PubMed  CAS  Google Scholar 

  • Xiao-Zhou Zhang, Zhong-Li Cui, Qing Hong, Shun-Peng Li (2005) High-Level Expression and Secretion of Methyl Parathion Hydrolase in Bacillus subtilis WB800. Appl Environ Microbiol 71:4101–4103

  • Wang PZ, Doi RH (1984) Overlapping promoters transcribed by Bacillus subtilis sigma55 and sigma37 RNA polymerase holoenzymes during growth and stationary phase. J Biol Chem 259:8619–8625

    PubMed  CAS  Google Scholar 

  • Wei ML, Webster DA, Stark BC (2000) Metabolic engineering of Sarratia marcescens with the bacterial hemoglobin gene alterations in fermentation pathways. Biotechnol. Bioengin 59:640–646

    Article  Google Scholar 

  • Yamamoto H, Serizawa M, Thompson J, Sekiguchi J (2001) Regulation of glv operon in Bacillus subtilis: YfiA(GLVR) is a positive regulator of the operon that is repressed through CcpA and cre. J Bacteriol 183:5110–5121

    Article  PubMed  CAS  Google Scholar 

  • Yuan G, Wong SL (1995) Regulation of groE expression in B.subtilis: the involvement of the sA-like promoter and the roles of the inverted repeat sequence (CIRCE). J Bacteriol 177:5427–5433

    PubMed  CAS  Google Scholar 

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Acknowledgements

We gratefully acknowledge the financial support of the National New Productions Project from the Science and Technology Ministry (P. R. China), and we thank the Bacillus Genetic Stock Centre of Ohio State University for generously providing the study materials.

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Authors and Affiliations

  1. Institute of Biochemical Engineering, College of Material Science and Chemical Engineering, Zhejiang University, Hangzhou, 310027, P.R. China

    Yang Ming-Ming & Cen Pei-Lin

  2. College of Animal Sciences, Northwest A&F University, Yangling, 712100, P.R. China

    Yang Ming-Ming

  3. Yangguang-Guangji Medical R&D co. Lt, Wuhan, 430062, P.R. China

    Yang Ming-Ming, Zhang Wei-Wei & Zhang Xi-Feng

Authors
  1. Yang Ming-Ming
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  2. Zhang Wei-Wei
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  3. Zhang Xi-Feng
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  4. Cen Pei-Lin
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Correspondence to Cen Pei-Lin.

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Ming-Ming, Y., Wei-Wei, Z., Xi-Feng, Z. et al. Construction and characterization of a novel maltose inducible expression vector in Bacillus subtilis . Biotechnol Lett 28, 1713–1718 (2006). https://doi.org/10.1007/s10529-006-9146-z

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  • DOI: https://doi.org/10.1007/s10529-006-9146-z

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