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Isolation and Molecular Screening of Glucansucrase Gene Harboring-Lactic Acid Bacteria
oleh: Ajitya Kurnia Hermawati, Mahardhika Hestiningtyas, Atiek Soemiati, Maksum R, Amarila Malik
Format: | Article |
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Diterbitkan: | Universitas Indonesia 2010-04-01 |
Deskripsi
Exopolysaccharides (EPS) have been possessed to be used in pharmaceutical, cosmetic and food industries. Lactic acid bacteria (LAB) produce a wide variety of exopolysaccharides and have been well reported carrying sucrase genes glucansucrase/ glucosyltransferase (gtf) and fructansucrase/fructosyltransferases (ftf), enzymes that are able to produce EPS. In this study, the isolation and screening of EPS producing-LAB (EPS-LAB) were carried out on modified de Mann-Rogosa-Sharpe (MRS) agar medium supplemented with 10% of sucrose on LAB isolated from various unique sugar containing-foods and -beverages originated from local sources. Besides obtaining EPS-LAB, this study aimed to screen for gtf gene as well as to molecular identify strains by using PCR technique. Degenerate primer pairs DegFor and DegRev which targeted the conserved region of gtf genes catalytic domain were used, whereas LABfw and LABrv were used to molecular identify strains using 16S rRNA gene. An approximately 660 base pairs (bp) amplicons which targeted gtf gene were obtained from 13 out of 16 srains chosen. Moreover, from PCR of 16S rRNA gene identification on gtf positive strains result, all strains were molecular identified as LAB after DNA sequencing analysis of 700 bp amplicons by using blastn. A rare EPS-producing LAB were obtain from both foods and beverages i.e. Weissella. Results revealed that strains obtained in this study are potential sources for exploring novel sucrase gene/s and obtain unique EPS polymer product/s.