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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)
  • 투고 : 2022.10.05
  • 심사 : 2022.12.20
  • 발행 : 2023.02.28

초록

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.

키워드

과제정보

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.

참고문헌

  1. Markets and Markets Research. 2020. Enzymes Market by Product Type (Industrial enzymes and Specialty enzymes), Source (Microorganism, Plant, and Animal), Type, Industrial Enzyme Application, Specialty Enzymes Application, and Region - (2019 - 2025). Available from https://www.marketsandmarkets.com/Market-Reports/enzyme-market-46202020.html/. Accessed July 29, 2022.
  2. Motyan JA, Toth F, Tozser, J. 2013. Research applications of proteolytic enzyme in molecular biology. Biomol. 3: 923-942. https://doi.org/10.3390/biom3040923
  3. Eatemadi A, Aiyelabegan HT, Negahdari B, Mazlomi MA, Daraee H, Daraee N, et al. 2017. Role of protease and protease inhibitors in cancer pathogenesis and treatment. Biomed. Pharmacol. 86: 221-231. https://doi.org/10.1016/j.biopha.2016.12.021
  4. Contesini FJ, de Melo RR, Sato HH. 2018. An overview of Bacillus proteases: from production to application. Crit. Rev. Biotech. 38: 321-334. https://doi.org/10.1080/07388551.2017.1354354
  5. Tufvesson P, Lima-Ramos J, Nordblad M, Lima-Ramos J, Nordblad M, Woodley JM. 2011. Guidelines and cost analysis for catalyst production in biocatalytic processes. Org. Process Res. Dev. 5: 266-274. https://doi.org/10.1021/op1002165
  6. Sharma KM, Kumar R, Panwar S, Kumar A. 2017. Microbial alkaline proteases: optimization of production parameters and their properties. J. Genet. Eng. Biotechnol. 15: 115-126. https://doi.org/10.1016/j.jgeb.2017.02.001
  7. Shafique T, Shafique J, Zahid S, Kazi M, Alnemer O, Ahmad A. 2021. Screening, selection and development of Bacillus subtilis aprIBL04 for hyper production of macromolecule alkaline protease. Saudi J. Biol. Sci. 28: 1494-1501. https://doi.org/10.1016/j.sjbs.2020.11.079
  8. Pundir1 RK, Rana S, Tyagi H. 2012. Studies on compatibility of fungal a alkaline protease with commercially available detergents. Int. J. Modern Biochem. 1: 41-56.
  9. Chung D, Yu W, Lim J, Kang N, Kwon Y, Choi G, et al. 2022. Characterization of the proteolytic activity of a halophilic Aspergillus reticulatus strain SK1-1 isolated from a solar saltern. Microorganisms 10: 29.
  10. Redes JL, Basu T, Ram-Mohan S, Ghosh CC, Chan EC, Sek AC, et al. 2019. Aspergillus fumigatus-Secreted alkaline protease 1 mediates airways hyperresponsiveness in severe asthma. Immunohorizons 3: 368-377 https://doi.org/10.4049/immunohorizons.1900046
  11. Benmrad MO, Moujehed E, Elhoul MB, Jaouadi NZ, Mechri S, Rekik H, et al. 2016. A novel organic solvent- and detergent-stable serine alkaline protease from Trametes cingulata strain CTM10101. Int. J. Biol. Macromol. 91: 961-972. https://doi.org/10.1016/j.ijbiomac.2016.06.025
  12. Jones EW. 1991. Three proteolytic systems in the yeast saccharomyces cerevisiae. J. Biol. Chem. 13: 7963-7966. https://doi.org/10.1016/S0021-9258(18)92922-4
  13. Chi Z, Ma C, Wang P, Li HF. 2007. Optimization of medium and cultivation conditions for alkaline protease production by the marine yeast Aureobasidium pullulans. Bioresour. Technol. 98: 534-538. https://doi.org/10.1016/j.biortech.2006.02.006
  14. Al-Askar AA, Rashad YM, Hafez EE, Abdulkhair WM, Baka ZA, Ghoneem KM. 2015. Characterization of alkaline protease produced by Streptomyces griseorubens E44G and its possibility for controlling Rhizoctonia root rot disease of corn. Biotechnol. Biotechnol. Equip. 29: 457-462. https://doi.org/10.1080/13102818.2015.1015446
  15. Al-Dhabi NA, Esmail GA, Ghilan AM, Arasu MV, Duraipandiyan V, Ponmurugan K. 2020. Characterization and fermentation optimization of novel thermo stable alkaline protease from Streptomyces sp. Al-Dhabi-82 from the Saudi Arabian environment for eco-friendly and industrial applications. J. King Saud Univer.-Sci. 32: 1258-1264. https://doi.org/10.1016/j.jksus.2019.11.011
  16. Bhunia B, Basak B, Dey A. 2012. A review on production of serine alkaline protease by Bacillus spp. J. Biochem. Tech. 3: 448-457. https://doi.org/10.1155/2012/905804
  17. Miao C, Han L, Lu Y, Feng H. 2020. Construvction of a high-expression system in Bacillus through transcriptomic profiling and promoter engineering. Microorganisms 8: e1030.
  18. Annamalaia N, Rajeswari MV, Balasubramanian T. 2014. Extraction, purification and application of thermostable and halostable alkaline protease from Bacillus alveayuensis CAS 5 using marine wastes. Food Bioprod. Process 9: 335-342.
  19. Lakshmi BKM, Muni Kumar D, Hemalatha KPJ. 2018. Purification and characterization of alkaline protease with novel properties from Bacillus cereus strain S8. J. Gen. Eng. Biotechnol. 16: 295-304. https://doi.org/10.1016/j.jgeb.2018.05.009
  20. Park HJ, Han SJ, Yim JH, Kim D. 2018. Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents. Korean J. Microbiol. 54: 60-68.
  21. Gopal N, Hill C, Ross PR, Beresford TP, Fenelon MA, Paul D. et al. 2015. The pevalence and control of Bacillus and related sporeforming bacteria in the dairy industry. Front. Microbiol. 6: e1418.
  22. Maurer K. 2004. Detergent protease. Curr. Opin. Biotechnol. 15: 330-334. https://doi.org/10.1016/j.copbio.2004.06.005
  23. Horikoshi K. 1991. Microorganisms in Alkaline Environments. Kodansha-VCH, Tokyo, Japan.
  24. Tsuchida O, Yamagata Y, Ishizuka T, Arai T, Yamada J, Takeuchi M, et al. 1986. An alkaline protease of an alkaline Bacillus sp. Curr. Microbiol. 14: 7-12. https://doi.org/10.1007/BF01568094
  25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265-275. https://doi.org/10.1016/S0021-9258(19)52451-6
  26. Kumar CG, Takagi H. 1999. Microbial alkaline proteases: from a bioindustrial viewpoint. Biotechnol. Adv. 17: 561-594. https://doi.org/10.1016/S0734-9750(99)00027-0
  27. Martinez R, Jakob F, Tu R, Siegert P, Maurer K, Schwaneberg U. 2013. Increasing activity and thermal resistance of Bacillus gibsonii alkaline protease (BgAP) by directed evolution. Biotechnol. Bioeng. 110: 711-720. https://doi.org/10.1002/bit.24766
  28. Deng A, Zhang G, Shi N, Wu J, Lu F, Wen T. 2014. Secretory expression, functional characterization, and molecular genetic analysis of novel halo-solvent-tolerant protease from Bacillus gibsonii. J. Microbiol. Biotechnol. 24: 197-208. https://doi.org/10.4014/jmb.1308.08094
  29. Rao CS, Sathish T, Pendyala B, Kumar TP, Prakasham RS. 2009. Development of a mathematical model for Bacillus circulans growth and alkaline protease production kinetics. J. Chem. Technol. Biotechol. 84: 302-307. https://doi.org/10.1002/jctb.2040
  30. Shafique T, Shafique J, Zahid S, Kazi M, Alnemer O, Ahmad A. 2021. Screening, selection and development of Bacillus subtilis aprIBL04 for hyper production of macromolecule alkaline protease. Saudi J. Bio. Sci. 28: 1494-1501. https://doi.org/10.1016/j.sjbs.2020.11.079
  31. Rao K, M. Narasu L. 2007. Alkaline protease from Bacillus firmus 7728. African J. Biotechnol. 6: 2493-2496. https://doi.org/10.5897/AJB2007.000-2395
  32. El-Gayar KE, Essa AM, Abada EA. 2020. Whey Fermentation for protease production using Bacillus thuringiensi isolated from mangrove rhizosphere soil in Jazan, Saudi Arabia. Pol. J. Environ. Stud. 29: 1-10.
  33. Yu JH, Jin HS, Choi WY, Yoon MH. 2006. Production and characterization of an alkaline protease from Bacillus licheniformis MH31. J. Appl. Biol. Chem. 49: 135-139.
  34. Lakshmi G, Prasad NN. 2015. Purification and characterization of alkaline protease from a mutant Bacillus licheniformis Bl8. Adv. Biol. Res. 9: 15-23.
  35. Cui H, Wang L, Yu Y. 2015. Production and characterization of alkaline protease from a high yielding and moderately halophilic strain of SD11 marine bacteria. J. Chem. e798304.
  36. Rekik H, Jaouadi NZ, Gargouri F, Bejar W, Frikha F, Jmal N, et al. 2019. Production, purification and biochemical characterization of a novel detergent-stable serine alkaline protease from Bacillus safensis strain RH12. Int. J. Biol. Macromol. 121: 1227-1239. https://doi.org/10.1016/j.ijbiomac.2018.10.139