Isolation, purification and biochemical characterization of alkaline α-amylase from Bacillus subtilis strain W3SFR5 isolated from kitchen Wastes
Applied Food Biotechnology,
Vol. 10 No 1 (2023),
3 janvier 2023
,
Page 9-19
https://doi.org/10.22037/afb.v10i1.39495
Résumé
Background and Objective: Amylase is a hydrolytic enzyme that breaks starch into simple sugars. This enzyme includes uses in starch production, brewery, detergent formulation, paper production and pharmaceuticals as a digest aid. The aim of the present study was to isolate, identify and characterize an alkaline amylase from bacteria of food wastes.
Material and Methods: Bacteria were isolated using serial dilution, screened on agar plates and characterized through biochemical assessments and 16S rRNA sequencing. After optimizing the bacterial growth conditions using one factor at a time method, the alkaline amylase was extracted from the culture broth and partially purified using Sephadex G-75 chromatography. Enzyme activity generated by submerged fermentation was assessed using 3,5-dinitrosalicylic and recorded as the mean of three replicates.
Results and Conclusion: The bacterial isolate W3SFR5 showed high amylolytic activity in agar culture. Biochemical analysis and sequencing of the 16S rRNA verified the bacterial isolate as Bacillus subtilis (GenBank accession number: OM258620). Bacillus subtilis W3SFR5 was propagated within 30–50 ℃ and pH 6-9. The partially purified Bacillus subtilis W3SFR5 amylase included a molecular weight of 65 kDa and demonstrated a maximum specific activity of 216.02U mg-1. The optimum temperature for the enzyme was 60 °C and the pH was 9. The W3SFR5 amylase was actively stable under temperatures of 50–70 °C and pH of 7-9. Furthermore, 5 mM Fe2+ increased W3SFR5 amylase activity. The enzyme was more resistant to organic solvents, surfactants, inhibitors and oxidizing agents than that most amylases were. Additionally, results showed that W3SFR5 amylase was compatible with most commercial detergents, indicating that it could be used as a detergent additive.
- ▪ Bacillus subtilis ▪ W3SFR5-amylase ▪ Biochemical characterization ▪ Detergent compatibility
Comment citer
Références
Ottoni JR, E Silva TR, de Oliveira VM, Zambrano Passarini MR. Characterization of amylase produced by cold-adapted bacteria from antarctic samples. Biocatal Agric Biotechnol. 2020; 23: -1-8https://doi.org/10.1016/j.bcab.2019.101452
Baweja M, Tiwari R, Singh PK, Nain L, Shukla P. An alkaline protease from Bacillus pumilus MP 27: Functional analysis of Its binding model toward Its applications as detergent additive. Front. Microbiol.2016; 7: 1-14https://doi.org/10.3389/fmicb.2016.01195.
Alves PDD, Siqueira FDF, Facchin S, Campolina C, Horta R, Victoria JMN, Kalapothakis E. Survey of microbial enzymes in soil, water and plant microenvironments. Open Microbiol J. 2014; 8: 25-31. https://doi.org/10.2174/1874285801408010025.
Ban X, Guo Y, Kaustubh B, Li C, Gu Z, Hu K, Li Z. The global amylase research trend in food science technology: A Data-driven analysis. Food Rev Int. 2021; 1-15. https://doi.org/10.1080/87559129.2021.1961267.
Menon G. Characterization of alkaliphilic, surfactant stable and raw starch digesting α-amylase from Bacillus subtilis Strain JS-16. J Microb Biochem T. 2014: 1-7https://doi.org/10.4172/1948-5948.s8-002.
Soumik Banerjee, Ghosh U. Production and characterization of glucoamylase by Aspergillus niger. Appl Food Biotechnol. 2017; 4: 19-26. https://doi.org/10.22037/afb.v4i1.13261.
Nigam PS. Microbial enzymes with special characteristics for biotechnological applications. Biomolecules. 2013; 3: 597-611.https://doi.org/10.3390/biom3030597.
Mitidieri S, Souza Martinelli AH, Schrank A, Vainstein MH. Enzymatic detergent formulation containing amylase from Aspergillus niger: A comparative study with commercial detergent formulations. Bioresour Technol. 2006: 97(10); 1217-1224.https://doi.org/10.1016/j.biortech.2005.05.022.
Singh S, Mangla J, Singh S. Evaluation of Aspergillus fumigatus NTCC1222 as a source of enzymes for detergent industry. Resourc Environ Sustain. 2021; 5l: .1-5
https://doi.org/10.1016/j.resenv.2021.100030.
Parmjit S. Panesar; Rupinder Kaur; Gisha Singla, Sangwan RS. Bio-processing of Agro-industrial Wastes for Production of Food-grade Enzymes: Progress and Prospects. Appl Food Biotechnol. 2016: 3; 208-227. https://doi.org/https://doi.org/10.22037/afb.v3i4.13458
Tiwari S, Shukla N, Mishra P, Gaur R. Enhanced production and characterization of a solvent stable amylase from solvent tolerant Bacillus tequilensis RG-01: Thermostable and surfactant resistant. Sci World J. 2014; 201: 1-11. https://doi.org/10.1155/2014/972763.
Kanimozhi M, Johny M, Gayathri N, Subashkumar R. Optimization and production of α -amylase from Halophilic Bacillus Species Isolated from Mangrove Soil Sources. Appl Environ Microbiol. 2014; 2: 70-73. https://doi.org/10.12691/jaem-2-3-2.
Kherouf M, Habbeche A, Benamia F, Saoudi B, Kerouaz B, Ladjama A. Statistical optimization of a novel extracellular alkaline and thermostable amylase production from thermophilic Actinomadura keratinilytica sp. Cpt29 and its potential application in detergent industry. Biocatal Agric Biotechnol. 2021; 35:1-11https://doi.org/10.1016/j.bcab.2021.102068.
Ritu Saini Saini HS, Dahiya A. Amylases characteristics and industrial applications. J Pharmacogn Phytochem.2017; 6: 1865-1871.
Niyomukiza S, Owino W, Maina JM, Maina N, Issifu M. Concomitant production of α-amylase and protease by Bacillus Aerius strain FPWSHA isolated from food wastes. Biointerface Res Appl Chem. 2022; 13:1-15https://doi.org/10.33263/BRIAC134.310.
Ibrahim ASS, Al-Salamah AA, Elbadawi YB, El-Tayeb MA, Ibrahim SSS. Production of extracellular alkaline protease by new halotolerant Alkaliphilic Bacillus sp. NPST-AK15 isolated from hyper saline soda lakes. Electron J Biotechnol. 2015; 18: 236-243.https://doi.org/10.1016/j.ejbt.2015.04.001.
Konkit M, Kim W. Activities of amylase, proteinase and lipase enzymes from Lactococcus chungangensis and its application in dairy products. J Dairy Sci. 2016; 99: 4999-5007. https://doi.org/10.3168/jds.2016-11002.
Al-Agamy MH, Alhuzani MR, Kelany MS, Hamed MM. Production and partial characterization of alpha-amylase enzyme from marine actinomycetes. BioMed Research International. 2021; 2021: 1-15https://doi.org/10.1155/2021/5289848.
Solanki P, Putatunda C, Kumar A, Bhatia R, Walia A. Microbial proteases: Ubiquitous enzymes with innumerable uses. 3 Biotech. 2021; 11: 1-25 https://doi.org/10.1007/s13205-021-02928-z.
Kizhakedathil MPJ, C, S.D. Acid stable alpha-amylase from Pseudomonas balearica VITPS19-Production, purification and characterization. Biotechnol Rep (Amst) 2021; 30:1-8
https://doi.org/10.1016/j.btre.2021.e00603.
Afrisham S, Badoei-Dalfard A, Namaki-Shoushtari A, Karami Z. Characterization of a thermostable, CaCl2-activated and raw-starch hydrolyzing alpha-amylase from Bacillus licheniformis AT70: Production under solid state fermentation by utilizing agricultural wastes. J Mol Catal B Enzym. 2016; 132: 98-106.https://doi.org/10.1016/j.molcatb.2016.07.002.
Thamy Livia Ribeiro Correa, Stella Karla dos Santos MOUTINHO, MARTIN, MLL, MARTINS, MA. Simultaneous α-amylase and protease production by the soil bacterium Bacillus sp. SMIA-2 under submerged culture using whey protein concentrate and corn steep liquor: Compatibility of enzymes with commercial detergents. Cienc Tecnol Aliment Campinas: 2011; 31: 843-848.
https://doi.org/10.1590/S0101-20612011000400003.
Hmidet N, Hadj Ali Nel, Haddar A, Kanoun S, Alya S-K, Nasri M. Alkaline proteases and thermostable α-amylase co-produced by Bacillus licheniformis NH1: Characterization and potential application as detergent additive. Biochem Eng J. 2009; 47: 71-79. https://doi.org/10.1016/j.bej.2009.07.005
Tamura K, Stecher G, Kumar S. MEGA11: Molecular evolutionary genetics analysis version 11. Mol Biol Evol. 2021; 38: 3022-3027. https://doi.org/10.1093/molbev/msab120.
Simair AA, Qureshi AS, Khushk I, Ali CH, Lashari S, Bhutto MA, Mangrio GS, Lu C. Production and partial characterization of alpha-amylase enzyme from Bacillus sp. BCC 01-50 and potential applications. BioMed Res Int. 2017; 2017:1-10.https://doi.org/10.1155/2017/9173040.
Bano S, Ul Qader SA, Aman A, Syed MN, Azhar A. Purification and characterization of novel alpha-amylase from Bacillus subtilis KIBGE HAS. AAPS Pharm Sci Tech. 2011; 12: 255-2261.https://doi.org/10.1208/s12249-011-9586-1.
Kishore Das, Doley R, Mukherjee AK. Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular α-amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India. Biotechnol Appl Biochem. 2004; 40: 291-298.
Promita Deb, Talukdar SA, Mohsina K, Sarke PK, Sayem A. Production and partial characterization of extracellular amylase enzyme from Bacillus amyloliquefaciens P-001.Springer Plus 2013; 2:154.https://doi.org/10.1186/2193-1801-2 154
Zhang, Jin-Wei, Zeng, Run-Ying. Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic J Mar Biotechnol. 2008. https://doi.org/10.1007/s10126-007-9035-z
Haddar A; Agrebi R, Bougatef A, Hmidet N, Sellami-Kamoun A, Nasri M. Two detergent stable alkaline serine-proteases from Bacillus mojavensis A21: purification, characterization and potential application as a laundry detergent additive. Bioresour Technol. 2009; 100: 3366-3373. https://doi.org/10.1016/j.biortech.2009.01.061
- Résumé ##plugins.themes.ojsPlusA.frontend.article.viewed##: 960 ##plugins.themes.ojsPlusA.frontend.article.times##
- pdf (English) ##plugins.themes.ojsPlusA.frontend.article.downloaded##: 662 ##plugins.themes.ojsPlusA.frontend.article.times##