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Enhancement of α-galactosidase production using novel Actinoplanes utahensis B1 strain: sequential optimization and purification of enzyme

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Abstract

α-Galactosidase is an important exoglycosidase belonging to the hydrolase class of enzymes, which has therapeutic and industrial potential. It plays a crucial role in hydrolyzing α-1,6 linked terminal galacto-oligosaccharide residues such as melibiose, raffinose, and branched polysaccharides such as galacto-glucomannans and galactomannans. In this study, Actinoplanes utahensis B1 was explored for α-galactosidase production, yield improvement, and activity enhancement by purification. Initially, nine media components were screened using the Plackett–Burman design (PBD). Among these components, sucrose, soya bean flour, and sodium glutamate were identified as the best-supporting nutrients for the highest enzyme secretion by A. Utahensis B1. Later, the Central Composite Design (CCD) was implemented to fine-tune the optimization of these components. Based on sequential statistical optimization methodologies, a significant, 3.64-fold increase in α-galactosidase production, from 16 to 58.37 U/mL was achieved. The enzyme was purified by ultrafiltration-I followed by multimode chromatography and ultrafiltration-II. The purity of the enzyme was confirmed by Sodium Dodecyl Sulphate–Polyacrylamide Agarose Gel Electrophoresis (SDS-PAGE) which revealed a single distinctive band with a molecular weight of approximately 72 kDa. Additionally, it was determined that this process resulted in a 2.03-fold increase in purity. The purified α-galactosidase showed an activity of 2304 U/mL with a specific activity of 288 U/mg. This study demonstrates the isolation of Actinoplanes utahensis B1 and optimization of the process for the α-galactosidase production as well as single-step purification.

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Acknowledgements

The authors express their gratitude to Dr. Harish Kumar Reddy, Manufacturing Science Department at Biological E Ltd for generously providing the laboratory to conduct the study. The authors would like to thank Dr. Sathish Thadikamala, Aurovaccines, Hyderabad, TS, India for his support in conducting statistical studiesas well as support during the drafting of this manuscript.

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Authors and Affiliations

  1. Department of Biotechnology, Vignan’s Foundation for Science, Technology and Research, Vadlamudi, Guntur District, Guntur, Andhra Pradesh, India

    K. Balumahendra, T. C. Venkateswarulu & D. John Babu

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  1. K. Balumahendra
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  2. T. C. Venkateswarulu
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  3. D. John Babu
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BK and JBD: conceptualized, designed and wrote the manuscript. VTC: revised the manuscript and also major contributor in writing the manuscript. All authors have read and approved the manuscript.

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Correspondence to D. John Babu.

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Balumahendra, K., Venkateswarulu, T.C. & Babu, D.J. Enhancement of α-galactosidase production using novel Actinoplanes utahensis B1 strain: sequential optimization and purification of enzyme. World J Microbiol Biotechnol 40, 91 (2024). https://doi.org/10.1007/s11274-023-03880-1

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