Isolation, Molecular Identification and Verification of Gene Encoding Bacterial Keratinase from Crocodile (Crocodylus porosus) Feces

4th International Conference on Biological Sciences and Biotechnology

Mamangkey, J.^1 ; Suryanto, D.^1 ; Munir, E.^1 ; Mustopa, A.Z.^2

Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia^1

Research Centre for Biotechnology, Indonesian Institute of Science (LIPI), Bogor, Indonesia^2

关键词: Aeromonas hydrophila;    Bacterial isolation;    Fragment lengths;    Gene-specific primers;    Keratinolytic bacteria;    Molecular approach;    Molecular identification;    Specific primers;   

Others  :  https://iopscience.iop.org/article/10.1088/1755-1315/305/1/012085/pdf
DOI  :  10.1088/1755-1315/305/1/012085

来源: IOP

【 摘 要 】

Keratinase is a group of protease enzymes which acts to degrade keratin. Keratin is a fiberous protein difficult to be degraded because of hydrogen and disulfide bonds. The purpose of this study was to isolate and to identify keratinase coding genes using molecular approach. The object of this study was crocodile feces from Asam Kumbang crocodile farm. Bacterial isolation was done by using feather agar (FA) and was screened in skim milk agar. Keratinolytic bacteria isolates were identified molecularly with 16S rDNA specific markers. Detection of keratinase gene was done by using keratinase-specific primers through primer3 application (version 0.4.0). The result showed that isolate FB3 are potential to produce keratinase. Isolate FB3 showed a clear zone in FA and SMA. FB3 was 100% similar to Aeromonas hydrophila. Amlification using kerD gene specific primer showed that FB3 was a novel bacteria possessing kerD gene with a fragment length of 750 bp. This result provided a new database of kerD gene.