Mutasi Gen xyncmu dan Pengaruhnya terhadap Stabilitas Enzim Xilanase CMU dari Bacillus halodurans

Abstract

Applications of xylanase enzyme on paper industry requires an enzyme characteristic through pH and resistant to high temperatures. A directed point mutations in site directed mutagenesis PCR which being the alternative to changing the enzyme’s characteristic and a several of amino acid on parental clon (M3) which effected with enzyme stability. This study aims to alter the stability of xylanase CMU and comparing the characters between CMU xylanase’s mutant and normal. The first step involves a plasmid with parental clones (Clone M3) which transferred to two pairs of mutagenic primers that Q53K and T194I. The amplicons from mutant plasmid pGEMalkxynaq1cmu M3 were transformed into Escherichia coli DH5α and selected with ampicillin media. Results of selection with ampicillin and restriction analysis were confirmed two positive mutant clones are Clone 4 and Clone 7 which brought of xyncmu gene. Gene mutation has succeeded to done as the result was Clone 7 which had enzyme’s characteristic through pH and resistant to high temperatures