1. Academic Validation
  2. Identification and characterization of a novel thermo-stable endo-β-N-acetylglucosaminidase from Rhizomucorpusillus

Identification and characterization of a novel thermo-stable endo-β-N-acetylglucosaminidase from Rhizomucorpusillus

  • J Biosci Bioeng. 2022 Oct;134(4):295-300. doi: 10.1016/j.jbiosc.2022.06.013.
Hanako Nishizawa 1 Mitsuhiro Iwamoto 2 Yasunori Ono 3
Affiliations

Affiliations

  • 1 Applied Microbiology Group, Biological Research Department, Daiichi Sankyo RD Novare Co., Ltd., 1-16-13 Kitakasai, Edogawa-ku, Tokyo 134-8630, Japan. Electronic address: nishizawa.hanako.rp@rdn.daiichisankyo.co.jp.
  • 2 Biologics Technology Research Laboratories, Biologics Division, Daiichi Sankyo Co., Ltd., 2716-1 Kurakake, Akaiwa, Chiyoda-machi, Gunma 370-0503, Japan.
  • 3 Applied Microbiology Group, Biological Research Department, Daiichi Sankyo RD Novare Co., Ltd., 1-16-13 Kitakasai, Edogawa-ku, Tokyo 134-8630, Japan.
Abstract

Endo-β-N-acetylglucosaminidase (ENGase) is an Enzyme that hydrolyzes the chitobiose core of N-glycans and is widely used for glycan analysis on glycoproteins and preparation of precursors for glycosylated compounds. While most of the ENGases that can hydrolyze complex-type glycans are derived from eukaryotes, their production by heterologous expression using Escherichia coli is insufficient, making the production process expensive. From an industrial perspective, there is a need for a less expensive Enzyme with higher activity and stability. In this study, we identified a novel ENGase gene from a thermophilic fungus, Rhizomucor pusillus, and named it Endo-Rp. Characterization of the recombinant Endo-Rp showed that the Enzyme had maximum hydrolytic activity at 60 °C and hydrolyzed high-mannose-type and biantennary complex-type glycans, but not (2,4)-branched triantennary complex-type or fucosylated glycans. Endo-Rp also hydrolyzed N-glycans attached to RNase B and human transferrin. In summary, we consider Endo-Rp to be a valuable Enzyme in various scientific and industrial applications.

Keywords

Endo-β-N-acetylglucosaminidase; Endoglycosidase; Glycoside hydrolase family 85; N-glycan; Rhizomucor pusillus.

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