Structure and cleavage pattern of a hyaluronate 3-glycanohydrolase in the glycoside hydrolase 79 family

Hao Huang; Xiaodong Hou; Ruirui Xu; Zhiwei Deng; Yang Wang; Guocheng Du; Yijian Rao; Jian Chen; Zhen Kang

doi:10.1016/j.carbpol.2021.118838

Structure and cleavage pattern of a hyaluronate 3-glycanohydrolase in the glycoside hydrolase 79 family

Carbohydr Polym. 2022 Feb 1:277:118838. doi: 10.1016/j.carbpol.2021.118838. Epub 2021 Nov 2.

Authors

Hao Huang¹, Xiaodong Hou², Ruirui Xu¹, Zhiwei Deng², Yang Wang¹, Guocheng Du¹, Yijian Rao³, Jian Chen⁴, Zhen Kang⁵

Affiliations

¹ The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China; The Science Center for Future Foods, Jiangnan University, Wuxi 214122, China.
² The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
³ The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: raoyijian@jiangnan.edu.cn.
⁴ The Science Center for Future Foods, Jiangnan University, Wuxi 214122, China; The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: jchen@jiangnan.edu.cn.
⁵ The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China; The Science Center for Future Foods, Jiangnan University, Wuxi 214122, China. Electronic address: zkang@jiangnan.edu.cn.

PMID: 34893255
DOI: 10.1016/j.carbpol.2021.118838

Abstract

Hyaluronidases have attracted a great deal of interest in the field of medicine due to their fundamental roles in the breakdown of hyaluronan. However, little is known about the catalytic mechanism of the hyaluronate 3-glycanohydrolases. Here, we report the crystal structure and cleavage pattern of a leech hyaluronidase (LHyal), which hydrolyzes the β-1,3-glycosidic bonds of hyaluronan. LHyal exhibits the typical structural features of glycoside hydrolase 79 family but contains a variable 'exo-pocket' loop where basic residues R102 and K103 are the structural determinants of hyaluronan binding. Through analysis of the hydrolysis of even- and odd-numbered hyaluronan oligosaccharides, we demonstrate that hexasaccharide is the shortest natural substrate, which can be cleaved from both the reducing and non-reducing ends to release disaccharides, and pentasaccharides are the smallest fragments for recognition and hydrolysis. These observations provide new insights into the degradation of hyaluronan and the evolutionary relationships of the GH79 family enzymes.

Keywords: GH79 family; Glycosaminoglycan; Hyaluronidase; Hydrolysis pattern; Leech.

MeSH terms

Animals
Hyaluronic Acid / chemistry
Hyaluronic Acid / metabolism*
Hyaluronoglucosaminidase / chemistry
Hyaluronoglucosaminidase / genetics
Hyaluronoglucosaminidase / metabolism*
Hydrolysis
Leeches / enzymology
Models, Molecular

Substances

Hyaluronic Acid
Hyaluronoglucosaminidase