Institute of Sericulture and System Biology, Southwest University, Chongqing, China.
Gene. 2013 Feb 1;514(1):62-8. doi: 10.1016/j.gene.2012.08.044. Epub 2012 Nov 29.
In this study, we report a novel cellulase [β-1,4-endoglucanase (EGase), EC 3.2.1.4] cDNA (Bh-EGase II) belonging to the glycoside hydrolase family (GHF) 45 from the beetle Batocera horsfieldi. The Bh-EGase II gene spans 720bp and consists of a single exon coding for 239 amino acid residues. Bh-EGase II showed 93.72% protein sequence identity to Ag-EGase II from the beetle Apriona germari. The GHF 45 catalytic site is conserved in Bh-EGase II. Bh-EGase II has three putative N-glycosylation sites at 56-58 (N-K-S), 99-101 (N-S-T), and 237-239 (N-Y-S), respectively. The cDNA encoding Bh-EGase II was expressed in baculovirus-infected insect BmN cells and Bombyx mori larvae. Recombinant Bh-EGase II from BmN cells and larval hemolymph had an enzymatic activity of approximately 928U/mg. The enzymatic catalysis of recombinant Bh-EGase II showed the highest activity at 50°C and pH6.0.
在这项研究中，我们报告了一种来自甲虫丽蝇（Batocera horsfieldi）的新型纤维素酶[β-1,4-内切葡聚糖酶（EGase），EC 3.2.1.4] cDNA（Bh-EGase II），它属于糖苷水解酶家族（GHF）45。Bh-EGase II 基因全长 720bp，由一个编码 239 个氨基酸残基的单一外显子组成。Bh-EGase II 与来自甲虫 Apriona germari 的 Ag-EGase II 的蛋白质序列具有 93.72%的同源性。GHF 45 催化位点在 Bh-EGase II 中保守。Bh-EGase II 有三个潜在的 N-糖基化位点，分别位于 56-58（N-K-S）、99-101（N-S-T）和 237-239（N-Y-S）。编码 Bh-EGase II 的 cDNA 在杆状病毒感染的昆虫 BmN 细胞和家蚕幼虫中表达。来自 BmN 细胞和幼虫血淋巴的重组 Bh-EGase II 的酶活性约为 928U/mg。重组 Bh-EGase II 的酶催化作用在 50°C 和 pH6.0 时表现出最高的活性。
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