Discovery of Highly Active Recombinant PNGase H+Variants Through the Rational Exploration of Unstudied Acidobacterial Genomes

被引：17

作者：

Guo, Rui-Rui ^{[1
]}

Comamala, Gerard ^{[2
]}

Yang, Huan-Huan ^{[1
]}

Gramlich, Marius ^{[3
]}

Du, Ya-Min ^{[1
]}

Wang, Ting ^{[1
]}

Zeck, Anne ^{[3
]}

Rand, Kasper Dyrberg ^{[2
]}

Liu, Li ^{[1
]}

Voglmeir, Josef ^{[1
]}

机构：

[1] Nanjing Agr Univ, Coll Food Sci & Technol, Glyc & Glycan Bioengn Res Ctr GGBRC, Nanjing, Peoples R China

[2] Univ Copenhagen, Dept Pharm, Prot Anal Grp, Copenhagen, Denmark

[3] Univ Tubingen, Nat & Med Sci Inst NMI, Reutlingen, Germany

来源：

FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY | 2020年 / 8卷

基金：

中国国家自然科学基金;

关键词：

acidic PNGase; N-glycans; glycoprotein deglycosylation; Dyella japonica; analytical glycoscience; TOOLS; AMIDASE;

D O I：

10.3389/fbioe.2020.00741

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Peptide-N-4-(N-acetyl-beta-glucosaminyl) asparagine amidases (PNGases,N-glycanases, EC 3.5.1.52) are indispensable tools in releasingN-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 speciesDyella japonica) 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 novelN-glycanase makes it a valuable tool for the analysis of protein glycosylation.

引用

页数：7

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