Protease of Stenotrophomonas sp. from Indonesian fermented food: gene cloning and analysis

  • Frans Kurnia Dexa Laboratory of Biomolecular Science, Dexa Medica, Cikarang, Indonesia.
  • Raymond R. Tjandrawinata Dexa Laboratory of Biomolecular Science, Dexa Medica, Cikarang, Indonesia.
  • Adi Yulandi Faculty of Biotechnology Atma Jaya Catholic University, Jakarta, Indonesia.
  • Maggy T. Suhartono | mthenawidjaja@yahoo.com Department of Food Science and Technology, Bogor Agricultural University, Bogor, Indonesia.

Abstract

Screening of proteolytic and fibrinolytic bacteria from Indonesian soy bean based fermented food Oncom revealed several potential isolates. Based on 16s rDNA gene analysis, one particular isolate with the highest proteolytic and fibrinolytic activity was identified as Stenotrophomonas sp. The protease gene was amplified to generate a 1749 bp Polymerase Chain Reaction product and BLAST analysis, revealed 90% homology with gene encoding protease enzyme from Stenotrophomonas maltophilia. The putative amino acid sequence indicated a serine protease enzyme with typical amino acid aspartate, histidine and serine in the catalytic triad. The gene was translated into a pre-pro-protein consisted of cleavage site on its N terminal and Pre-Peptidase Cterminal domain. Cloning of the protease gene in pET22b with Escherichia coli BL21 DE3 as the host showed that the gene was expressed as insoluble protein fraction. This is the first report for analysis of protease gene from food origin Stenotrophomonas sp.

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Published
2018-02-23
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Original Articles
Keywords:
Protease gene, Stenotrophomonas sp., Indonesian fermented food Oncom, serine protease
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How to Cite
Kurnia, F., Tjandrawinata, R. R., Yulandi, A., & Suhartono, M. T. (2018). Protease of Stenotrophomonas sp. from Indonesian fermented food: gene cloning and analysis. Journal of Biological Research - Bollettino Della Società Italiana Di Biologia Sperimentale, 90(2). https://doi.org/10.4081/jbr.2017.6428