PRODUCTION AND CHARACTERIZATION OF COLLAGENASE FROM INDONESIAN Staphylococcus sciuri

• Main Author: Satria Wira Kusuma, Arif
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45360
• Institution: Institut Teknologi Bandung

Collagen is one of main protein present in fish skin, such as common snakehead fish (Channa striata). Collagenase is an endopeptidase that can digest collagen into peptides. Peptides obtained from collagen digestion by collagenase have been reported to have immunomodulator, antioxidant, antihypertension, antiproliferation, and antimicrobial activity. The aim of this research was to extract and characterize collagen from C. striata skin wastes, to select and identify collagenase producing bacteria, and also to produce and characterize collagenase. Collagen from C. striata was extracted by using acid-soluble method and then characterized based on its molecular weight. Collagenase producing bacteria were selected based on their extracelluler protease caseinolytic and collagenolytic activities. Collagenase producing bacteria identification was done by using 16s rRNA PCR method. Molecular identification result was then being confirmed with morphologic analysis by using Gram staining method, catalase test, and bacteria inoculation in Vogel Johnsson Agar (VJA) and Mannitol Salt Agar (MSA) selective medium. Collagenase production was done by using skim milk enriched medium. Free cells supernatant obtained from the production process was concentrated. Supernatant concentrate was then characterized by SDS-PAGE and zymography methods. Collagenase stability and activity were then being tested against temperature and pH. From 32.36 gram of C. striata skin, 7.08 mg collagen with molecular weight characteristic of 68.39, 73.13, and 93.37 kDa was obtained. Collagenase producing bacterium was identified as Staphylococcus sciuri. Collagenase from S. sciuri had molecular weight characteristic of 14.62 kDa. From 100 mL production scale 845.5 µg collagenase was obtained. Collagenase from S. sciuri was stabile and had activity at the temperature range of 37–80 ºC and pH range of 3–11.