Assay of Proteolytic Activity of Bacillus Subtilis for Collagen Isolation from Snakehead Fish (Channa striata) Scales

Year 2023, Volume: 40 Issue: 3, 515 - 521, 30.09.2023

Siti Nurkasanah

Abstract

The collagen source could be derived from fish scales like snakehead fish (Channa striata). One of the possible methods can be used enzymatic methods. The objective of this research was to isolate and characterize the collagen from C. striata scale using B. subtilis protease. This study was an experimental method utilizing the protease enzyme produced by B. subtilis. The proteolytic activity of B. subtilis was seen from the ability of the bacteria to produce a clear zone on skim-casein agar and inoculated into tryptic soy broth (TSB) as a submerged medium to produce protease. The collagen isolation using B. subtilis protease (crude and freeze-dried enzyme) with ratios 1:1, 1:10, 1:100, and 1:1000 and incubation for 6, 12, 18, and 24 hours. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to determine the molecular weight (MW) of collagen proteins, protein determination by Bradford methods, and morphology was observed by scanning electron microscopy (SEM). All data were analyzed using using SPSS.16 software for windows. The highest collagen was at 12 hours (1:100) incubation using freeze-dried enzymes resulting in a mass of 0.10 grams and a protein content of 3.17 g/ml (p<0.05). SDS-PAGE and SEM visualization showed the collagen might be classified as type I collagen consisting of two chains (α1 and α2) with molecular weights approaching 118.03 kDa and 112.20 kDa. Collagen from Channa striata scales can be isolated enzymatically using a protease produced by B. subtilis and can be characterized as type 1 collagen.

Keywords

Bacillus subtilis, Channa striata, collagen, protease, scale

Supporting Institution

Grants DPP/SPP Faculty of Mathematics and Natural Sciences, University of Brawijaya, Indonesia at 2015.

Project Number

2015

Thanks

The authors are sincerely grateful to the laboratory facilities at the Faculty of Mathematics and Natural Sciences, University of Brawijaya

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