通过对未研究的酸杆菌基因组进行合理探索发现高活性重组PNGase H变体
Discovery of Highly Active Recombinant PNGase H Variants Through the Rational Exploration of Unstudied Acidobacterial Genomes.
作者信息
Guo Rui-Rui, Comamala Gerard, Yang Huan-Huan, Gramlich Marius, Du Ya-Min, Wang Ting, Zeck Anne, Rand Kasper Dyrberg, Liu Li, Voglmeir Josef
机构信息
Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, China.
Protein Analysis Group, Department of Pharmacy, University of Copenhagen, Copenhagen, Denmark.
出版信息
Front Bioeng Biotechnol. 2020 Jul 3;8:741. doi: 10.3389/fbioe.2020.00741. eCollection 2020.
Peptide- -(-acetyl-β-glucosaminyl) asparagine amidases (PNGases, -glycanases, EC 3.5.1.52) are indispensable tools in releasing -glycans from glycoproteins. So far, only a limited number of PNGase candidates are available for the structural analysis of glycoproteins and their glycan moieties. Herein, a panel of 13 novel PNGase H candidates (the suffix H refers to the acidic pH optimum of these acidobacterial PNGases) was tested in their recombinant form for their deglycosylation performance. One candidate (originating from the bacterial species ) showed superior properties both in solution-phase and immobilized on amino-, epoxy- and nitrilotriacetate resins when compared to currently acidic available PNGases. The high expression yield compared to a previously described PNGase H, broad substrate specificity, and good storage stability of this novel -glycanase makes it a valuable tool for the analysis of protein glycosylation.
肽-（-乙酰基-β-葡糖胺基）天冬酰胺酶（PNGases，-聚糖酶，EC 3.5.1.52）是从糖蛋白中释放-聚糖的不可或缺的工具。到目前为止，仅有有限数量的PNGase候选物可用于糖蛋白及其聚糖部分的结构分析。在此，测试了一组13种新型PNGase H候选物（后缀H指这些酸杆菌PNGases的最适酸性pH）的重组形式的去糖基化性能。与目前可用的酸性PNGases相比，一种候选物（源自细菌物种）在溶液相以及固定在氨基、环氧和次氮基三乙酸树脂上时均表现出优异的性能。与先前描述的PNGase H相比，该新型-聚糖酶的高表达产量、广泛的底物特异性和良好的储存稳定性使其成为蛋白质糖基化分析的有价值工具。
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