来源Paenibacillus sp.的β-环糊精葡萄糖基转移酶基因的克隆及在大肠杆菌中的表达

Clone and expression of β-cyclodextrin glycosyltransferase from Paenibacillus sp. in Escherichia coli.

以1株产β-环糊精葡萄糖基转移酶(β-Cyclodextrin Glycosyltransferase,简称β-CGTase)的嗜碱性Paenibacillus sp.XW-6-66的染色体为模板,通过PCR扩增获得编码β-CGTase的基因。序列分析表明该基因长度为2139 bp,是1个编码713个氨基酸的完整阅读框。将该基因克隆至表达型载体pET28a(+)后,转入大肠杆菌BL21(DE3)中高效表达,37℃摇瓶培养,11 h酶活力达到2938.2 U/mL,证实了高效表达的可能性,为进一步实现该β-CGTase的工业化生产奠定了基础。

Recombinant beta-cyclodextrin glycosyltransferase（β-CGTase） was obtained by cloning the PCR gene fragment from alkalophilic Paenibacillus sp.strain.XW-6-66.The nucleotide sequence of the recombinant β-CGTase contained an open reading frame of 2139 base pairs encoding 713 amino acid residues.The nucleotide sequence was ligated into pET28a（＋） vector and then transformed into Escherichia coli BL21（DE3） for the expression.The β-CGTase activity reached to 2938.2 U/mL after 11 h incubation at 37 ℃.This provided a possibility for high-level expression for β-CGTase and further realized the industrial production of β-CGTase to lay a foundation.