Optimization and molecular identification of PUA-14 bacterial isolate from protease-producing mangrove waters

Abstract

Abstract. Protease enzymes are hydrolase enzymes that break down proteins into simpler molecules and are extensively utilized in the food, pharmaceutical, and other chemical industries. Selecting microorganisms as enzyme producers can address the high demand for enzymes and promote sustainable production. This research analyzed carbon sources, nitrogen sources, trace elements, carbon source concentrations, nitrogen source concentrations, and optimum inoculum concentrations for PUA-14 bacterial isolates producing protease. In addition, biomolecular analysis was conducted, including DNA isolation, amplification, and sequencing, which aimed to identify protease-producing PUA-14 bacterial isolates from mangrove waters using biomolecular-based identification. The research employed experimental techniques, specifically the One Factor at A Time (OFAT) method and Response Surface Methodology (RSM) with a Central Composite Design (CCD) using Design Expert 13 software. Results demonstrated that PUA-14 bacterial isolates had optimal conditions for producing protease. These conditions included a carbon source of lactose at 0.5%, a nitrogen source of NaNO3 at 2.5%, trace element Zn, and an inoculum concentration of 2.5%. Molecular-based analysis revealed that isolate PUA-14 showed similarities with Bacillus pseudomycoides strain LB-AsDX1-3. The research findings are anticipated to offer novel insights into the regulation of protease activity by the PUA-14 bacteria and establish the groundwork for the creation of more effective industrial applications.