Bioproduction of alpha-amylase from Bacillus amyloliquefasciens using taro (Colocasia esculenta) as a medium

Joliesa Mae S. Toledo; Simon G. Alcantara; Darlon V. Lantican; Abegail Papong; Nadine Janelle Sinco; Sharmaine Daya; Susan M. Mercado

Authors

Joliesa Mae S. Toledo Institute of Biological Sciences, Central Mindanao University, Maramag, 8710, Philippines; Institute of Biological Sciences, University of the Philippines Los Banos, Laguna, 4030, Philippines
Simon G. Alcantara Institute of Biological Sciences, University of the Philippines Los Banos, Laguna, 4030, Philippines
Darlon V. Lantican Institute of Biological Sciences, University of the Philippines Los Banos, Laguna, 4030, Philippines
Abegail Papong Institute of Biological Sciences, University of the Philippines Los Banos, Laguna, 4030, Philippines
Nadine Janelle Sinco Institute of Biological Sciences, University of the Philippines Los Banos, Laguna, 4030, Philippines
Sharmaine Daya Food and Feed Laboratory, National Institute of Molecular Biology and Biotechnology, University of the Philippines-Los Banos, Laguna, 4030, Philippines
Susan M. Mercado Food and Feed Laboratory, National Institute of Molecular Biology and Biotechnology, University of the Philippines-Los Banos, Laguna, 4030, Philippines

Keywords:

amylase, Bacillus amyloliquefaciens, Colocasia esculenta, RSM

Abstract

Amylases are among the most important industrial enzymes, with diverse applications ranging from conversion of starch into sugar syrups to the production of cyclodextrins for the pharmaceutical industry. Bacillus amyloliquefaciens NBRC 15535 can be a potential source of amylase production. The use of taro (Calocasia esculenta) as primary starch submerged for α-amylase production and scale up contributes as substrate for industrial enzyme production. Report on taro as fermentation substrate are scarce despite its high starch content and availability in tropical regions like Philippines. This study aimed to optimize alpha-amylase production conditions for large-scale industrial applications. Stat-Ease Design-Expert software was employed to design the bioproduction parameters, specifically pH, temperature, and substrate concentration. For the scale-up process, a two-level factorial design was utilized to determine the optimal fermentation conditions. Based on small-scale experimental results, the optimum conditions were identified as pH 8.0, 50°C, and a 4% substrate concentration. Statistical analysis demonstrated that temperature was the only significant factor influencing α-amylase production (p < 0.0001), while pH, substrate concentration, and their interactions were not significant. Scale-up experiments revealed that conditions favoring biomass accumulation did not necessarily correspond to maximal specific enzyme activity, highlighting the importance of distinguishing cell growth from enzyme production. Time-course analyses indicated thatthe culture remained in theexponential growth phase within 24 h of fermentation, suggesting that the true optimum for α-amylase production was not yet reached under the evaluated conditions. Therefore, extending the incubation period to 48–72 h and exploring broader pH ranges are proposed to capture potential growth-associated or stability-driven increases in enzyme yield. Overall, this study establishes baseline process parameters and demonstrates the feasibility of taro-based media for scalable α-amylase production to meetindustrydemand, while providing aclear framework for future optimization. under the evaluated conditions. Therefore, extending the incubation period to 48–72 h and exploring broader pH ranges are proposed to capture potential growth-associated or stability-driven increases in enzyme yield. Overall, this study establishes baseline process parameters and demonstrates the feasibility of taro-based media for scalable α-amylase production to meet industry demand, while providing a clear framework for future optimization.

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