藤黄节杆菌β-1,3-葡聚糖酶基因在大肠杆菌中的克隆及表达

Cloning and Expression of the β-1,3-Glucanase from Arthrobacter luteus in Escherichia coli

以藤黄节杆菌基因组为模板,克隆获得β-1,3-葡聚糖酶基因,分别构建至原核表达载体pET-28a(+)与pBAD18上,诱导获得高效表达,粗酶液对酵母多糖的裂解活性达到161 U/mL。在裂解酵母试验中,粗酶液也表现出较高的活性。研究中得到一株突变株,其对于研究β-1,3-葡聚糖酶在裂解酵母中多糖结合域的地位和作用有着重要的价值。

The gene coding β-1,3-glucanase in Arthrobacter luteus genome was cloned into prokaryotic vector pET-28a（ ＋ ） and pBAD18, and the heterogeneous protein was successfully expressed in the recombinant Escherichia coli. The endoglucanase activity of crude extract to lyse Zymosan A was about 161 U/mL. Also, the viable yeast cells could be lysed by the crude extract effectively. Furthermore, we obtained a mutant strain that would be valuable for studying the roles of β-1,3-glucanase＇s carbohydrate binding modules （CBM13） in viable yeast-lyric by enzymic method.