Optimization of α-amylase Production from a Novel Bacterial Strain, Geobacillus kaustophilus Strain AN11, Employing Response Surface Methodology

Author: Asama Nadaf and Nasir R. Shaikh

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Abstract

To overcome numerous industrial barriers, it is critical to investigate thermostable α-amylase-producing bacteria in demanding conditions. This study aimed to identify and characterize α-amylase-producing bacteria isolated from hot springs. The isolates were identified as Geobacillus kaustophilus strain AN11 based on their molecular characterization. An optimization technique based on statistical experimental design is applied to increase thermostable α-amylase production by a thermotolerant Geobacillus kaustophilus strain AN11 isolate. The pH, temperature, salt (NaCl) level, and nitrogen supply (tryptophan) were found to have a substantial influence on enzyme production using the one variant at a time approach. The response surface methodology (RSM) was used to determine the optimal process conditions among the variables picked. A Box-Behnken design was utilized to develop a polynomial quadratic model that correlated the link between the four variables and α-amylase activity. The optimum culture variables for α-amylase synthesis were pH 5, temperature 40oC, tryptophan 0.6 gm, and NaCl 2.52 gm. Temperature studies demonstrated that the crude enzyme had the highest activity and stability at 60, 65, and 70°C. The temperature and α-amylase production had a linear relationship. Thus, molecular identification of this novel strain Geobacillus kaustophilus strain AN11 isolates, purification, and production of amylases are proposed for effective application in various industries. Industrial detergent's effectiveness to wash clothes might be enhanced by adding enzymes to the detergent. This study has shown that these thermostable bacteria are a great supply of amylase that may be employed commercially to sustain economic activity.

Keywords

α-amylase, Response surface methodology, Industries, Enzymes, Box-Behnken

Conclusion

Amylases account for thirty percent of the world's total requirement for enzymes and find applications in the paper, health care, and cleaning product industries, amongst many others. The newly identified bacterial strain Geobacillus kaustophilus strain AN11 can be employed to produce amylase because it is a potentially affordable source for the enzyme. Thermophilic amylase can be produced at a relatively low incubation temperature (60oC), and the enzyme was thermostable after being exposed to 70oC for 72 hours. RSM statistical software is quite helpful in scientific investigations comprising the identification of the key factors or the responses of these key factors on the multi-factor dependent variables. Based on the RSM findings, the enzyme activity was at its peak at a pH of 5 and temperature 40°C. In this study, we found that altering the nitrogen source resulted in the highest enzyme activity. Concentrations of tryptophan greater than 0.6 gm are not ideal for growth. Enzyme activity peaked at a concentration of 0.4 gm tryptophan. The optimal salt concentration for enzyme activity was 2.52 gm. As a follow-up investigation to validate the techno-feasibility of its mass commercial production, further method validation for the scaled-up production techniques is preferred.

References

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How to cite this article

Asama Nadaf and Nasir R. Shaikh (2023). Optimization of α-amylase Production from a Novel Bacterial Strain, Geobacillus kaustophilus Strain AN11, Employing Response Surface Methodology. Biological Forum – An International Journal, 15(5): 988-993.