Kloning Gen Xilanase Termozim Dari Streptomyces costaricanus 45I-3

Abstract

Biopolymer xylan has a great potential to produce useful end product such as xylose. Xylanases is a group of enzyme that has ability to hydrolyse xylan or polymer of xyloses and xylooligosaccharides. Complete hydrolysis of xylans involves the synergistic action of a xylanolytic enzymes, including ß-xylanase, ß-xylosidase, -glucuronidase, -L-arabinofuranosidase, and acetyl xylan esterase. The aims of this research were to isolate xylanase gene from Streptomyces costaricanus 45I-3 and to clone this gene into competence Escherichia coli DH5 . Genomic DNA was obtained from S. costaricanus 45I-3 that have known has ability to produce thermostabile extracellular xylanase. Genomic DNA was partially digested with restriction enzyme Sau3A1. Genomic DNA fragments of size 4-10 kb were inserted at BamHI site in pBluescript II KS (+) vector and transformed into E. coli DH5 . Based on blue-white selection approximately 12000 recombinant colonies were obtained. The presence of xylanase gene was identified by screening employing congored plate clearance assay. One positive clone containing the xylanase gene was confirmed and was designated as pHS11. Preliminary characterization of pHS11 using EcoRI showed that it contained a 6,0 kb DNA insert. Xylanase were produced has xylanase spesific activity 6,393 U/mg, -xylosidase activity 0,208 U/ml, arabinofuranosidase activity 0,079 U/ml and acetyl xylan esterase activity 0,063 U/ml as well.