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All Journal Jurnal Pembelajaran dan Biologi Nukleus Bioscientist : Jurnal Ilmiah Biologi
Mellanie Alia Putri
Universitas Andalas
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Immunology & microbiology Languange, Linguistic, Communication & Media Social Sciences

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Screening of Metalloprotease-Producing Thermophilic Bacteria and Optimization of Trace Elements for Enzyme Activity Mellanie Alia Putri; Anthoni Agustien; Yetty Marlida; Ayumi Rizci Puspita
JURNAL PEMBELAJARAN DAN BIOLOGI NUKLEUS Vol 12, No 2: Jurnal Pembelajaran Dan Biologi Nukleus June 2026
Publisher : Universitas Labuhanbatu

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.36987/jpbn.v12i2.9385

Abstract

Background: Thermophilic bacteria are potential sources of thermostable enzymes with important applications in biotechnology and industry. Among these enzymes, metalloproteases are of particular interest because their catalytic activity depends on metal ions. This study aimed to screen thermophilic bacterial isolates from hot springs in Solok Regency, West Sumatra, Indonesia, for metalloprotease production and to evaluate the effects of trace elements on metalloprotease activity. Methodology: Four thermophilic bacterial isolates (TUA-104,                   TUA-106, TUA-109, and TUA-113) were cultivated in protease production medium (pH 8.0) at 50oC for 24 hours, and crude enzymes were obtained by centrifugation. Metalloprotease screening was conducted using EDTA inhibition assays through well diffusion and specific protease activity analyses. The isolate showing the strongest metalloprotease characteristics was further evaluated using 5 ppm Ca²⁺, Mn²⁺, and Zn²⁺ supplementation. Findings: The results showed that isolates TUA-106, TUA-109, and TUA-113 exhibited characteristics of metalloprotease-producing bacteria. Among them, TUA-106 showed the most consistent inhibition pattern, with specific activity decreasing from 0.780 U/mg in the control to 0.522 U/mg after treatment with 10 mM EDTA. Zn²⁺ produced the highest specific activity (0.284 U/mg), followed by Mn²⁺ (0.263 U/mg), whereas Ca²⁺ resulted in the lowest activity (0.036 U/mg). Contribution: These findings indicate that TUA-106 is a potential Zn-dependent metalloprotease-producing thermophilic bacterium and provide new insight into the metalloprotease-producing potential of thermophilic bacteria from Solok geothermal environments for future biotechnological applications
Isolation and Characterization of Protease-Producing Thermophilic Bacteria from Padang Dama Hot Spring, Solok, Indonesia Mellanie Alia Putri; Anthoni Agustien; Yetti Marlida; Feskaharny Alamsjah
Bioscientist : Jurnal Ilmiah Biologi Vol. 14 No. 2 (2026): June
Publisher : Department of Biology Education, FSTT, Mandalika University of Education, Indonesia.

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33394/bioscientist.v14i2.20118

Abstract

This study aimed to isolate, screen, and partially characterize protease-producing thermophilic bacteria from the Padang Dama hot spring, Solok, Indonesia. The main issue addressed in this study is the limited information regarding thermophilic bacteria capable of producing protease enzymes from this geothermal environment, despite their potential use in industrial processes requiring stable enzymes. Isolation was carried out using Nutrient Agar followed by incubation at 50 °C for 24 hours. Screening of protease activity was performed using Skim Milk Agar based on clear zone formation, and the proteolytic index (PI) was calculated to evaluate enzyme activity. Selected isolates were further characterized through macroscopic, microscopic, and biochemical analyses. The results showed that three isolates exhibited proteolytic activity, with isolate TUA-109 showing the highest proteolytic index, indicating superior enzymatic potential compared to other isolates. Partial characterization revealed that isolate TUA-109 is a Gram-negative, rod-shaped bacterium with positive catalase activity and motility, which are important traits supporting its adaptability and potential for enzyme production under extreme conditions. These findings highlight the potential of thermophilic bacteria from the Padang Dama hot spring as sources of protease enzymes, with isolate TUA-109 identified as a promising candidate for further investigation, particularly for industrial enzyme production. Future studies are recommended to optimize enzyme production and evaluate its application in industrial processes.
Co-Authors Anthoni Agustien Ayumi Rizci Puspita Feskaharny Alamsjah Yetti Marlida Yetty Marlida