IMPROVEMENT OF ALPHA-AMYLASE ACTIVITY FROM BACILLUS LICHENIFORMIS USING UV RADIATION AND MODIFIED MEDIA COMPOSITION

Authors

  • M. Mahfuza Khatun Dept. of Genetic Engineering & Biotechnology, Bangabandhu Sheikh Mujibur Rahman Maritime University, Bangladesh
  • Farahdiba Zarin Dept. of Biomedical Engineering, Military Institute of Science and Technology (MIST), Dhaka
  • Kazi Zarin Tasnim Dept. of Biomedical Engineering, Military Institute of Science and Technology (MIST), Dhaka
  • Fahria Sarmin Dept. of Biomedical Engineering, Military Institute of Science and Technology (MIST), Dhaka

DOI:

https://doi.org/10.3329/brc.v9i2.67092

Keywords:

Alpha-amylase, Enzyme, Mutation, UV irradiation, Bacillus licheniformis

Abstract

Alpha-amylases are glycosidic bond hydrolyzing enzymes produced in many microorganisms. In the modern era, the spectrum of amylase applications has expanded into various industries, including medical fields. Among other approaches, strain improvement by mutation and culture optimization are attractive methods for increasing the productivity of alpha-amylase in bacterial strains. The present study aimed at enhancing the yield of the enzyme alpha-amylase from the host strain Bacillus licheniformis. The UV irradiation has been used to induce mutations in the bacterial strain with different time durations. Afterwards, the mutant strains were cultured in different modified growth media for enzyme production. Among the four time periods,the  highest enzyme activity (705.23 U/ml) was observed after 24 hours which is more than two-fold higher compared to the enzyme activity of the host stain (345.31 U/ml). In addition, enzyme yield varied significantly in modified culture media (i.e, MP1 and MP2, MP3 and MP4) with different carbon and nitrogen compositions. The mutant strain cultured in MP3 medium had  the highest amylase activity which was 1461.2 U/ml at 24 hours. More interestingly, this medium also had the highest enzyme activity (1080.5 U/ml) at 48 hours. The trend of enzyme activity showed a decreasing tendency when the strains cultured in MP1, MP2, and MP3 media, but exceptionally, a sharp decreasing trend was seen in MP4 media from 622.1 U/ml at 24 hours to 428.4 U/ml at 48 hours. From the findings, it can be concluded that B. licheniformis is a good candidate to employ for alpha-amylase production in a modified MP3 culture condition for a  short time. However, other limiting factors need to be considered for increasing the yield.

References

Aida, M. F. & Hanan, M. A. N. (2016), Production, optimization and characterization of extracellular amylase from halophilic Bacillus lichineformis AH214. African Journal of Biotechnology, 15, 670-683.

Al-Johani, N. B., Al-Seeni, M. N. & Ahmed, Y. M. (2017), Optimization of alkaline alpha-amylase production by thermophilic Bacillus subtilis. Afr J Tradit Complement Alternative Medicine, 14, 288-301.

Arora, N. K., Mishra, J. & Mishra, V. (2020), Microbial enzymes: roles and applications in industries, Springer.

Welker, N. E. and Campbell, L. L. (1963), Effect of carbon sources on formation of a-amylase by Bacillus stearothermophilus. Journal of bacteriology, 86, 681-686.

Dash, B. K., Rahman, M. M. & Sarker, P. K. (2015), Molecular identification of a newly isolated Bacillus subtilis BI19 and optimization of production conditions for enhanced production of extracellular amylase. BioMed Research International, 859805.

Dike P. E., O. C. J., Akaranta O. and Oji A. (2022), Isolation and identification of amylase-producing microorganisms. GSC Biological and Pharmaceutical Sciences 20, 076-082.

He, H., Yu, Q., Ding, Z., Zhang, L., Shi, G. & Li, Y. (2023), Biotechnological and food synthetic biology potential of platform strain: Bacillus licheniformis. Synthetic and Systems Biotechnology, 8, 281-291.

Hiteshi, K., Didwal, G. & Gupta, R. (2016), Production optimization of alpha-amylase from Bacillus licheniformis. Journal of Advance Research in Business Management and Accounting (ISSN: 2456-3544), 2, 01-14.

Hormoznejad, R., Saberi, S., Moridnia, A., Azish, M. & Elyasi F. B. (2022), Optimization of the alpha-Amylase production from microbial source: A systematic review of experimental studies. Trends in Medical Sciences, 2.

Ikram-ul-haq, S., Saleem, A. & Javed, M. (2009), Mutagenesis of Bacillus licheniformis through ethyl methane sulfonate for alpha amylase production. Pak. Journal of Botany, 41, 1489-1498.

Khandaker M. Khalid-Bin-Ferdaus, M. F. H., Sheikh A. M. S., & Ahmed S., (2018), Commercial production of alpha amylase enzyme for potential use in the textile industries in Bangladesh. International Journal of Biosciences (IJB), 13, 149-157.

Miller, G. L. (1959), Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical chemistry, 31, 426-428.

Mohammad, B. T., Al Daghistani, H. I., Jaouani, A., Abdel-latif, S. & Kennes, C. (2017), Isolation and characterization of thermophilic bacteria from Jordanian hot springs: Bacillus licheniformis and thermomonas hydrothermalis isolates as potential producers of thermostable enzymes. International Journal of Microbiology, 6943952.

Niu, D., Zuo, Z., Shi, G. Y. & Wang, Z. X. (2009), High yield recombinant thermostable alpha-amylase production using an improved Bacillus licheniformis system. Microbial Cell Factory, 8, 58.

Periasamy A., Subash. C. B. G., Bidur P. C., & Lakshmipriya, A. T. (2017), Microbial enzymes and their applications in industries and medicine 2016. BioMed Research International, Hindawi Publishing Corporation.

Rahman, S. M. A. S. R. (2008), Production and partial characterization of extracellular α-Amylase by Trichoderma viride. Bangladesh J Microbiology 25, 99-103.

Sankaralingam S., Ramasubburayan, T. S., Prakash S. R. and Kumar C. (2012), Optimization of culture conditions for the production of Amylase from Bacillus licheniformis on submerged fermentation. American-Eurasian Journal of Agriculture & Environmental Science, 12 1507-1513.

Surendra S., Vinni. S., Monohar. L. S. (2011), Biotechnological applications of industrially important amylase enzyme. International Journal of Phrma and Biosciences, 2, 486-496.

Wang, J. R., LI, Y. Y., Liu, D. N., Liu, J. S., Li, P., Chen, L. Z. & Xu, S. D. (2015), Codon Optimization significantly improves the expression level of alpha -Amylase gene from Bacillus licheniformis in Pichia pastoris. Biomedical Research International, 248680.

Yao, D., Su, L., Li, N. & Wu, J. (2019), Enhanced extracellular expression of Bacillus stearothermophilus alpha-amylase in Bacillus subtilis through signal peptide optimization, chaperone overexpression and alpha-amylase mutant selection. Microial Cell Factory, 18, 69.

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Published

21-06-2023

How to Cite

Khatun, M. M., Zarin, F., Tasnim, K. Z., & Sarmin, F. (2023). IMPROVEMENT OF ALPHA-AMYLASE ACTIVITY FROM BACILLUS LICHENIFORMIS USING UV RADIATION AND MODIFIED MEDIA COMPOSITION. Bioresearch Communications - (BRC), 9(2), 1351–1356. https://doi.org/10.3329/brc.v9i2.67092

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