Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Alkaline Protease Production from Bacillus gibsonii 6BS15-4 Using Dairy Effluent and Its Characterization as a Laundry Detergent Additive

  • Polson Mahakhan (Department of Microbiology, Faculty of Science, Khon Kaen University) ;
  • Patapee Apiso (Department of Microbiology, Faculty of Science, Khon Kaen University) ;
  • Kannika Srisunthorn (Department of Microbiology, Faculty of Science, Khon Kaen University) ;
  • Kanit Vichitphan (Department of Biotechnology, Faculty of Technology, Khon Kaen University) ;
  • Sukanda Vichitphan (Department of Biotechnology, Faculty of Technology, Khon Kaen University) ;
  • Sukrita Punyauppa-path (Department of Mathematics and Science, Faculty of Agriculture and Technology, Rajamangala University of Technology Isan Surin Campus) ;
  • Jutaporn Sawaengkaew (Department of Microbiology, Faculty of Science, Khon Kaen University)
  • Received : 2022.10.05
  • Accepted : 2022.12.20
  • Published : 2023.02.28
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Abstract

Protease is a widely used enzyme particularly in the detergent industry. In this research, we aimed to isolate alkaline protease-producing bacteria for characterization as a laundry detergent additive. The screening of alkaline protease production was investigated on basal medium agar plus 1% skim milk at pH 11, with incubation at 30℃. The highest alkaline protease-producing bacterium was 6BS15-4 strain, identified as Bacillus gibsonii by 16S rRNA gene sequencing. While the optimum pH was 12.0, the strain was stable at pH range 7.0-12.0 when incubated at 45℃ for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60℃ and the enzyme was stable at 55℃ when incubated at pH 11.0 for 60 min. Metal ions K+, Mg2+, Cu2+, Na+, and Zn2+ exhibited a slightly stimulatory effect on enzyme activity. The enzyme retained over 80% of its activity in the presence of Ca2+, Ba2+, and Mn2+. Thiol reagent and ethylenediaminetetraacetic acid did not inhibit the enzyme activity, whereas phenylmethylsulfonyl fluoride significantly inhibited the protease activity. The alkaline protease from B. gibsonii 6BS15-4 demonstrated efficiency in blood stain removal and could therefore be used as a detergent additive, with potential for various other industrial applications.

Keywords

Acknowledgement

This research was supported by the Plant Genetic Project under The Royal Initiative of Her Royal Highness Princess Maha Chakri Sirindhorn and Research and Graduate Studies, Khon Kaen University, Thailand.

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