Cold Active Amylase Production from <i>Bacillus cereus</i> RGUJS2023

http://www.doi.org/10.26538/tjnpr/v7i11.20

Authors

  • Amrita Samanta Department of Microbiology, Faculty of science, Raiganj University, Raiganj, 733134, West Bengal, India.
  • Satyasundar Pradhan Department of Microbiology, Faculty of science, Raiganj University, Raiganj, 733134, West Bengal, India.
  • Subhas C. Jana Department of Microbiology, Faculty of science, Raiganj University, Raiganj, 733134, West Bengal, India.

Keywords:

Starch hydrolysis, Growth kinetics, Cold-active alpha amylase, Characterization, Bacillus cereus

Abstract

Cold-active enzymes have significant biotechnological prospects and provide a number of ecological and economic advantages by reducing the heating cost. The present investigation describes the production and characterization of a novel cold-active enzyme from the RGUJS2023 strain of Bacillus cereus. This enzyme would be used for the benefit of the food, starch, and pharmaceutical industries. The growth kinetics of this strain showed that starch utilization with reducing sugar production was within 4 hours of the exponential growth of the strain with the initial production of enzyme. The starch was fully utilized in the media within 6 hours of culture growth; however, the produced reducing sugar remained same up to 20 hours of the cell’s growth. The highest production of the enzyme was in the late exponential to early stationary phase. This strain produced the highest enzyme after incubating for 16 hours at 37°C and at pH 6.9. This enzyme was characterized as cold-active alpha-amylase. This enzyme showed maximum activity with 2% starch solution at 28°C and at pH 6.5 after 30 min of incubation and was stable at room temperature (30°C) for a long time. Cold active enzyme activity was stimulated in the presence of Mn2+, Mg2+ and Sn2+ and inhibited in the presence of Pb2+ and Cu2+. After a careful consideration of the above finding, it may be considered that this cold-active alpha-amylase from Bacillus cereus RGUJS2023 is an outstanding, affordable, and reliable choice for industrial applications.  

References

Chakraborty S, Khopade A, Biao R, Jian W, Liu XY, Mahadik K, Chopade B, Zhang L, Kokare C. Characterization and stability studies on surfactant, detergent and oxidant stable α-amylase from marine haloalkaliphilic Saccharopolyspora sp. A9. J Mol Catal B Enzym. 2011; 68(1): 52-58.

Zaferanloo B, Bhattacharjee S, Ghorbani MM, Mahon PJ, Palombo EA. Amylase production by Preussia minima, a fungus of endophytic origin: optimization of fermentation conditions and analysis of fungal secretome by LC-MS. BMC Microbiol. 2014; 14: 1-12.

Kalsoom F, Anjum FR, Anam S, Khan S, Hasan F. Effect of temperature, pH and metal ions on amylase produced from selected indigenous extremophile bacteria in Pakistan. Int J Biosci. 2018; 13(03): 262-275.

Farooq MA, Ali S, Hassan A, Tahir HM, Mumtaz S, Mumtaz S. Biosynthesis and industrial applications of α-amylase: A review. Arch Microbiol. 2021; 203: 1281-1292.

Jujjavarapu SE, Dhagat S. Evolutionary trends in industrial production of α-amylase. Recent Pat Biotechnol. 2019; 13(1): 4-18.

Gopinath SC, Anbu P, Arshad MM, Lakshmipriya T, Voon CH, Hashim U, Chinni SV. Biotechnological processes in microbial amylase production. Biomed Res. Int. 2017; 2017.

Arabacı N, Arıkan B. Isolation and characterization of a cold-active, alkaline, detergent stable α-amylase from a novel bacterium Bacillus subtilis N8. Prep Biochem. 2018; 48(5): 419-426.

Ray RC, Kar S, Nayak S, Swain MR. Extracellular α-amylase production by Bacillus brevis MTCC 7521. Food Biotechnol. 2008; 22(3): 234-246.

Zhang JW, Zeng RY. Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic. Mar Biotechnol. 2008; 10: 75-82.

Yadav AN, Sachan SG, Verma P, Kaushik R, Saxena AK. Cold active hydrolytic enzymes production by psychrotrophic Bacilli isolated from three sub‐glacial lakes of NW Indian Himalayas. J Basic Microbiol. 2016; 56(3): 294-307.

Kuddus M, Ahmad IZ. Cold-active extracellular α-amylase production from novel bacteria Microbacterium foliorum GA2 and Bacillus cereus GA6 isolated from Gangotri glacier, Western Himalaya. J Genet Eng Biotechnol. 2012; 10(1): 151-159.

Dou S, Chi N, Zhou X, Zhang Q, Pang F, Xiu Z. Molecular cloning, expression, and biochemical characterization of a novel cold-active α-amylase from Bacillus sp. dsh19-1. Extremophiles. 2018; 22: 739-749.

Shinde VK, Vamkudoth KR. Maltooligosaccharide forming amylases and their applications in food and pharma industry. J Food Sci Technol. 2021; 27: 1-12.

Adrio JL, Demain AL. Microbial enzymes: tools for biotechnological processes. Biomolecules. 2014; 4(1): 117-139.

Hamid B, Bashir Z, Yatoo AM, Mohiddin F, Majeed N, Bansal M, Poczai P, Almalki WH, Sayyed RZ, Shati AA, Alfaifi MY. Cold-active enzymes and their potential industrial applications—A Review. Molecules. 2022;27(18): 5885.

Poddar A, Gachhui R, Jana SC. Optimization of physico-chemical condition for improved production of hyperthermostable β amylase from Bacillus subtilis DJ5. J Biochem Technol. 2012; 3(4): 370-374.

Ray RR, Jana SC, Nanda G. β-Amylase from Bacillus megaterium. Folia Microbiol. 1994; 39: 567-570.

Ray RR, Jana SC, Nanda GE. Optimization of physico-chemical conditions for improved production of beta-amylase by Bacillus megaterium B6. Acta Microbiol. Pol. 1995; 44(1).

Ranjan K, Lone MA, Sahay S. Detergent compatible cold-active alkaline amylases from Clavispora lusitaniae CB13. J. Microbiol. Biotechnol. Food Sci. 2016; 5(4): 306.

Evurani SA, Berebon PD, Eze OC, Emencheta SC, Anyim G, Okpalanwa CF. Purification and Characterization of α-Amylase from Bacillus cereus SM22 Isolated from Deteriorating Cocoyam. Trop J Nat Prod Res. 2022; 6(2): 219-226.

Sethi BK, Nanda PK, Sahoo S, Sena S. Characterization of purified α-amylase produced by Aspergillus terreus NCFT 4269.10 using pearl millet as substrate. Cogent food agric. 2016; 2(1): 1158902.

Mishra S, Behera N. Amylase activity of a starch degrading bacteria isolated from soil receiving kitchen wastes. Afr J Biotechnol. 2008; 7(18): 3326-3331..

Doss A, Anand SP. Purification and characterization of extracellular amylolytic enzyme from Aspergillus species. Afr J Biotechnol. 2012; 11(83): 14941- 14945.

Suganthi C, Mageswari A, Karthikeyan S, Gothandam KM. Insight on biochemical characteristics of thermotolerant amylase isolated from extremophile bacteria Bacillus vallismortis TD6 (HQ992818). Microbiol. 2015; 84: 210-218.

Zhang JW, Zeng RY. Psychrotrophic amylolytic bacteria from deep sea sediment of Prydz Bay, Antarctic: diversity and characterization of amylases. World J Microbiol Biotechnol. 2007; 23: 1551-1557.

Zhang JW, Zeng RY. Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic. Mar. Biotechnol. 2008; 10: 75-82.

Mihaela C, Teodor N, Gabriela B, Peter S. Cold adapted amylase and protease from new Streptomyces 4 Alga Antarctic strain. Innov Rom Food Biotechnol. 2009; 10(5): 23-30.

Abo-Kamer AM, Abd-El-salam IS, Mostafa FA, Mustafa AE, Al-Madboly LA. A promising microbial α-amylase production, and purification from Bacillus cereus and its assessment as antibiofilm agent against Pseudomonas aeruginosa pathogen. Microb Cell Factories. 2023; 22(1): 141.

Msarah MJ, Ibrahim I, Hamid AA, Aqma WS. Optimization and production of alpha amylase from thermophilic Bacillus spp. and its application in food waste biodegradation. Heliyon. 2020; 6(6).

Qin Y, Huang Z, Liu Z. A novel cold-active and salt-tolerant α-amylase from marine bacterium Zunongwangia profunda: molecular cloning, heterologous expression and biochemical characterization. Extremophiles. 2014; 18: 271-281.

Liu J, Zhang Z, Dang H, Lu J, Cui Z. Isolation and characterization of a cold-active amylase from marine Wangia sp. C52. Afr J Microbiol Res. 2011; 5: 1156-1162.

Alva S, Anupama J, Savla J, Chiu YY, Vyshali P, Shruti M, Yogeetha BS, Bhavya D, Purvi J, Ruchi K, Kumudini BS. Production and characterization of fungal amylase enzyme isolated from Aspergillus sp. JGI 12 in solid state culture. Afr J Biotechnol. 2007; 6(5): 576.

Ottoni JR, e Silva TR, de Oliveira VM, Passarini MR. Characterization of amylase produced by cold-adapted bacteria from Antarctic samples. Biocatal. Agric. Biotechnol. 2020; 23: 101452.

Halder D, Biswas E, Basu M. Amylase production by Bacillus cereus strain BRSC-S-A26MB under optimal laboratory condition. Int. J. Curr. Microbiol. App. Sci. 2014; 3(6):1035-1047.

Elechi DP, Jason OC, Onyewuchi A, Akuma O. Optimization of B. cereus PW4 alpha amylase production by OVAT technique. GSC biol. pharm. sci. 2022; 20(1): 83-90.

Abo-Kamer AM, Abd-El-salam IS, Mostafa FA, Mustafa AE, Al-Madboly LA. A promising microbial α-amylase production, and purification from Bacillus cereus and its assessment as antibiofilm agent against Pseudomonas aeruginosa pathogen. Microbial Cell Factories. 2023 Aug 1;22(1):141

Published

2023-12-01

How to Cite

Samanta, A., Pradhan, S., & Jana, S. C. (2023). Cold Active Amylase Production from <i>Bacillus cereus</i> RGUJS2023: http://www.doi.org/10.26538/tjnpr/v7i11.20. Tropical Journal of Natural Product Research (TJNPR), 7(11), 5172–5177. Retrieved from https://tjnpr.org/index.php/home/article/view/3014