Alkaline Protease Production from Bacillus gibsonii 6BS15-4 Using Dairy Effluent and Its Characterization as a Laundry Detergent Additive
1
조회
0
다운로드
1
인용
0
참고문헌
키워드
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°C. 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°C for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60°C and the enzyme was stable at 55°C 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.
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°C. 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°C for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60°C and the enzyme was stable at 55°C 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.
참고문헌 (36)
- "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)" (2020)[URL]
- Motyan JA, "Research applications of proteolytic enzyme in molecular biology", Biomol, 3, pp.923~942 (2013)
- Eatemadi A, "Role of protease and protease inhibitors in cancer pathogenesis and treatment", Biomed. Pharmacol, 86, pp.221~231 (2017)
- Contesini FJ, "An overview of Bacillus proteases: from production to application", Crit. Rev. Biotech, 38, pp.321~334 (2018)
- Tufvesson P, "Guidelines and cost analysis for catalyst production in biocatalytic processes", Org. Process Res. Dev, 5, pp.266~274 (2011)
- Sharma KM, "Microbial alkaline proteases: optimization of production parameters and their properties", J. Genet. Eng. Biotechnol, 15, pp.115~126 (2017)
- Shafique T, "Screening, selection and development of Bacillus subtilis apr-IBL04 for hyper production of macromolecule alkaline protease", Saudi J. Biol. Sci, 28, pp.1494~1501 (2021)
- Pundir RK, "Studies on compatibility of fungal a alkaline protease with commercially available detergents", Int. J. Modern Biochem, 1, pp.41~56 (2012)
- Chung D, "Characterization of the proteolytic activity of a halophilic Aspergillus reticulatus strain SK1-1 isolated from a solar saltern", Microorganisms, 10, pp.29~ (2022)
- Redes JL, "Aspergillus fumigatus–Secreted alkaline protease 1mediates airways hyperresponsiveness in severe asthma", Immunohorizons, 3, pp.368~377 (2019)
- Benmrad MO, "A novel organic solvent- and detergent-stable serine alkaline protease from Trametes cingulata strain CTM10101", Int. J. Biol. Macromol, 91, pp.961~972 (2016)
- Jones EW, "Three proteolytic systems in the yeast saccharomyces cerevisiae", J. Biol. Chem, 13, pp.7963~7966 (1991)
- Chi Z, "Optimization of medium and cultivation conditions for alkaline protease production by the marine yeast Aureobasidium pullulans", Bioresour. Technol, 98, pp.534~538 (2007)
- Al-Askar AA, "Characterization of alkaline protease produced by Streptomyces griseorubens E44G and its possibility for controlling Rhizoctonia root rot disease of corn", Biotechnol. Biotechnol. Equip, 29, pp.457~462 (2015)
- Al-Dhabi NA, "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, pp.1258~1264 (2020)
- Bhunia B, "A review on production of serine alkaline protease by Bacillus spp", J. Biochem. Tech, 3, pp.448~457 (2012)
- Miao C, "Construvction of a high-expression system in Bacillus through transcriptomic profiling and promoter engineering", Microorganisms, 8, pp.e1030~ (2020)
- Annamalaia N, "Extraction, purification and application of thermostable and halostable alkaline protease from Bacillus alveayuensis CAS 5 using marine wastes", Food Bioprod. Process, 9, pp.335~342 (2014)
- Lakshmi BKM, "Purification and characterization of alkaline protease with novel properties from Bacillus cereus strain S8", J. Gen. Eng. Biotechnol, 16, pp.295~304 (2018)
- 박하주, "Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents", 미생물학회지, 54(1), pp.60~68 (2018), 한국미생물학회[DOI]
- Gopal N, "The pevalence and control of Bacillus and related sporeforming bacteria in the dairy industry", Front. Microbiol, 6, pp.e1418~ (2015)
- Maurer K, "Detergent protease", Curr. Opin. Biotechnol, 15, pp.330~334 (2004)
- Horikoshi K, "Microorganisms in Alkaline Environments" (1991), Kodansha-VCH
- Tsuchida O, "An alkaline protease of an alkaline Bacillus sp", Curr. Microbiol, 14, pp.7~12 (1986)
- Lowry OH, "Protein measurement with the Folin phenol reagent", J. Biol. Chem, 193, pp.265~275 (1951)
- Kumar CG, "Microbial alkaline proteases: from a bioindustrial viewpoint", Biotechnol. Adv, 17, pp.561~594 (1999)
- Martinez R, "Increasing activity and thermal resistance of Bacillus gibsonii alkaline protease (BgAP) by directed evolution", Biotechnol. Bioeng, 110, pp.711~720 (2013)
- Aihua Deng, "Secretory Expression, Functional Characterization, and Molecular Genetic Analysis of Novel Halo-Solvent-Tolerant Protease from Bacillus gibsonii", Journal of Microbiology and Biotechnology, 24(2), pp.197~208 (2014), 한국미생물·생명공학회[DOI]
- Rao CS, "Development of a mathematical model for Bacillus circulans growth and alkaline protease production kinetics", J. Chem. Technol. Biotechol, 84, pp.302~307 (2009)
- Shafique T, "Screening, selection and development of Bacillus subtilis apr-IBL04 for hyper production of macromolecule alkaline protease", Saudi J. Bio. Sci, 28, pp.1494~1501 (2021)
- Rao K, "Alkaline protease from Bacillus firmus 7728", African J. Biotechnol, 6, pp.2493~2496 (2007)
- El-Gayar KE, "Whey Fermentation for protease production using Bacillus thuringiensi isolated from mangrove rhizosphere soil in Jazan, Saudi Arabia", Pol. J. Environ. Stud, 29, pp.1~10 (2020)
- Jeong-Hyeon Yu, "Production and Characterization of an Alkaline Protease fromBacillus licheniformis MH31", Journal of Applied Biological Chemistry, 49(4), pp.135~139 (2006), 한국응용생명화학회
- Lakshmi G, "Purification and characterization of alkaline protease from a mutant Bacillus licheniformis Bl8", Adv. Biol. Res, 9, pp.15~23 (2015)
- Cui H, "Production and characterization of alkaline protease from a high yielding and moderately halophilic strain of SD11 marine bacteria", J. Chem, pp.e798304~ (2015)
- Rekik H, "Production, purification and biochemical characterization of a novel detergent-stable serine alkaline protease from Bacillus safensis strain RH12", Int. J. Biol. Macromol, 121, pp.1227~1239 (2019)