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High-level expression, purification and characterization of a recombinant medium-temperature α-amylase from Bacillus subtilis

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Abstract

Alpha-amylases are important industrial enzymes with a wide range of applications. Although medium-temperature alpha amylase (AmyE) has some practical advantages, its low yield has limited its applications. When an amyE gene from Bacillus subtilis BF768 was cloned into vector pWB980 and over-expressed in B. subtilis WB600, high activities (723 U ml−1) of secreted AmyE were produced. Recombinant AmyE was purified to a specific activity of 36 U mg−1 having optimal activity at pH 6.0 and 60°C.

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References

  • Ehrlich SD (1977) Replication and expression of plasmids from Staphylococcus aureus in Bacillus subtilis. Proc Natl Acad Sci USA 70:1680–1682

    Article  Google Scholar 

  • Harwood CR (1992) Bacillus subtilis and its relatives: molecular biological and industrial workhorses. Trends Biotechnol 10:247–256

    Article  CAS  PubMed  Google Scholar 

  • Liu YH, Lu FP, Li Y, Wang JL, Gao C (2008a) Acid stabilization of Bacillus licheniformis alpha amylase through introduction of mutations. Appl Microbiol Biotechnol 80:795–803

    Article  CAS  PubMed  Google Scholar 

  • Liu YH, Lu FP, Li Y, Yin XB, Wang Y, Gao C (2008b) Characterisation of mutagenised acid-resistant alpha-amylase expressed in Bacillus subtilis WB600. Appl Microbiol Biotechnol 78:85–94

    Article  CAS  PubMed  Google Scholar 

  • NSCFFI (National Standardization Center for Food and Fermentation Industry) (2006) General methods of determination for industrial enzymes. In: The food industry standard of People’s Republic of China (Fermentation product volume). Standards Press of People’s Republic of China, Beijing. pp 117–135

  • Richardson TB, Tan XQ, Frey G, Callen W, Cabell M, Lam D, Macomber J, Short JM, Robertson DE, Miller C (2002) A novel, high performance enzyme for starch liquefaction. J Biol Chem 277:26501–26507

    Article  CAS  PubMed  Google Scholar 

  • Sivaramakrishnan S, Gangadharan D, Nampoothiri KM, Soccol CR, Pandey A (2006) Alpha-amylases from microbial sources—an overview on recent developments. Food Technol Biotechnol 44:173–184

    CAS  Google Scholar 

  • Wang PZ, Doi RH (1984) Overlapping promoters transcribed by Bacillus subtilis σ55 and σ37 RNA polymerase holoenzymes during growth and stationary phase. J Biol Chem 259:8619–8625

    CAS  PubMed  Google Scholar 

  • Wu SC, Wong SL (1999) Development of improved pUB110-based vectors for expression and secretion studies in Bacillus subtilis. J Biochem 72:185–195

    CAS  Google Scholar 

  • Wu XC, Lee W, Tran L, Wong SL (1991) Engineering a Bacillus subtilis expression–secretion system with a strain deficient in six extracellular proteases. J Bacteriol 173:4558–4952

    CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin, 300457, People’s Republic of China

    Yihan Liu & Fuping Lu

  2. College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China

    Yihan Liu, Fuping Lu, Jing Sun, Yu Li, Jianling Wang & Jing Xiao

  3. Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, T6G 2P5, Canada

    Guanqun Chen & Crystal L. Snyder

Authors
  1. Yihan Liu
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  2. Fuping Lu
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  3. Guanqun Chen
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  4. Crystal L. Snyder
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  5. Jing Sun
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  6. Yu Li
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  7. Jianling Wang
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  8. Jing Xiao
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Correspondence to Fuping Lu.

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Liu, Y., Lu, F., Chen, G. et al. High-level expression, purification and characterization of a recombinant medium-temperature α-amylase from Bacillus subtilis . Biotechnol Lett 32, 119–124 (2010). https://doi.org/10.1007/s10529-009-0112-4

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  • DOI: https://doi.org/10.1007/s10529-009-0112-4

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