원문정보
초록
영어
A β-1,3-glucanase was widely used in various biotechnological processes and overproduction of its was required for versatile industrial utilization. We subcloned β-1,3-glucanase gene (exgA) from Aspergillus oryzae and examined overexpression of β-1,3-glucanase by using δ sequence-mediated integration. We constructed plasmid, pRSδH-exgA for repetitive integration of exgA gene. This plasmid contains ADH1 promoter for constitutive expression, exoinulinase signal sequence for secretion, loxP-CgHIS3-loxP selective marker for repetitive selection and δ target sequence for integration of β-1,3-glucanase. The pRSδH-exgA plasmid was transformed into S. cerevisiae BY4742△exg1 strain and transformant showing β-1,3-glucanase activity was selected. Subsequently, the CgHIS3 marker was removed for marker recycling and the pRSδH-exgA plasmid was repeatedly transformed to induce overexpression of β-1,3-glucanase. By repetitive transformation, the activity of β-1,3-glucanase was sequentially increased and these results suggested that δ-mediated integration was efficient for enhancement of β-1,3-glucanase activity.