Optimization and Molecular Identification of Protease-Producing Thermophilic Bacterial Isolate TUA-26

Abstract

Protease enzymes are used as biocatalysts that can accelerate a reaction. Thermostable enzymes produced by thermophilic bacteria are needed in the industrial field because of their ability to withstand high temperatures. The thermophilic bacterial isolate TUA-26 that produces protease was to analyze the optimum conditions of temperature, pH, agitation, trace elements, carbon sources, and nitrogen sources; the stability of protease enzymes; and identify the species based on analysis of 16S rRNA sequences. The research method was conducted experimentally using the Response Surface Methodology (RSM) type of Central Composite Design (CCD) in software statistic design expert 13. The optimum conditions for the thermophilic bacterial isolate TUA-26 to produce protease at 70°C, 8 pH, 150 rpm, Zn trace element, glucose carbon source, and NaNO₃ nitrogen source. Protease activity produced by the thermophilic bacterial isolate TUA-26 increased before and after optimization. The protease enzyme stability test still lasts up to 80% for 7 h from the initial activity. Molecular identification using 16S rRNA of isolate TUA-26 that has similarities with Brevibacillus borstelensis strain UTM105. The thermophilic bacterial isolate TUA-26 that produces protease can be improved through optimization by engineering the extrinsic environment and producing thermostable enzymes, which is of great interest for commercial and industrial use in both food and non-food fields.