Biochemical Characterization of Two Rhamnogalacturonan Lyases From Bacteroides ovatus ATCC 8483 With Preference for RG-I Substrates

Weiyang Wang; Yibing Wang; Haoting Yi; Yang Liu; Guojing Zhang; Le Zhang; Kevin H. Mayo; Ye Yuan; Yifa Zhou

doi:10.3389/fmicb.2021.799875

Biochemical Characterization of Two Rhamnogalacturonan Lyases From Bacteroides ovatus ATCC 8483 With Preference for RG-I Substrates

Weiyang Wang, Yibing Wang, Haoting Yi, Yang Liu, Guojing Zhang, Le Zhang, Kevin H. Mayo, Ye Yuan, Yifa Zhou

Chemical and Structural Biology (TMED)

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

Rhamnogalacturonan lyase (RGL) cleaves backbone α-1,4 glycosidic bonds between L-rhamnose and D-galacturonic acid residues in type I rhamnogalacturonan (RG-I) by β-elimination to generate RG oligosaccharides with various degrees of polymerization. Here, we cloned, expressed, purified and biochemically characterized two RGLs (Bo3128 and Bo4416) in the PL11 family from Bacteroides ovatus ATCC 8483. Bo3128 and Bo4416 displayed maximal activity at pH 9.5 and pH 6.5, respectively. Whereas the activity of Bo3128 could be increased 1.5 fold in the presence of 5 mM Ca²⁺, Bo4416 required divalent metal ions to show any enzymatic activity. Both of RGLs showed a substrate preference for RG-I compared to other pectin domains. Bo4416 and Bo3128 primarily yielded unsaturated RG oligosaccharides, with Bo3128 also producing them with short side chains, with yields of 32.4 and 62.4%, respectively. Characterization of both RGLs contribute to the preparation of rhamnogalacturonan oligosaccharides, as well as for the analysis of the fine structure of RG-I pectins.

Original language	English (US)
Article number	799875
Journal	Frontiers in Microbiology
Volume	12
DOIs	https://doi.org/10.3389/fmicb.2021.799875
State	Published - Jan 11 2022

Bibliographical note

Funding Information:
National Natural Science Foundation of China (No. 32000907); Jilin Provincial Department of Science and Technology, Medicine and Health Project (No. 20200504005YY); and the Scientific and Technologic Foundation of Jilin Province (Nos. 20200201004JC and 20200708059YY).

Funding Information:
National Natural Science Foundation of China (No. 32000907); Jilin Provincial Department of Science and Technology, Medicine and Health Project (No. 20200504005YY); and the Scientific and

Publisher Copyright:
Copyright © 2022 Wang, Wang, Yi, Liu, Zhang, Zhang, Mayo, Yuan and Zhou.

Keywords

Bacteroides ovatus
polysaccharide lyase family 11
rhamnogalacturonan lyase
rhamnogalacturonan oligosaccharides
type I rhamnogalacturonan

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title = "Biochemical Characterization of Two Rhamnogalacturonan Lyases From Bacteroides ovatus ATCC 8483 With Preference for RG-I Substrates",

abstract = "Rhamnogalacturonan lyase (RGL) cleaves backbone α-1,4 glycosidic bonds between L-rhamnose and D-galacturonic acid residues in type I rhamnogalacturonan (RG-I) by β-elimination to generate RG oligosaccharides with various degrees of polymerization. Here, we cloned, expressed, purified and biochemically characterized two RGLs (Bo3128 and Bo4416) in the PL11 family from Bacteroides ovatus ATCC 8483. Bo3128 and Bo4416 displayed maximal activity at pH 9.5 and pH 6.5, respectively. Whereas the activity of Bo3128 could be increased 1.5 fold in the presence of 5 mM Ca2+, Bo4416 required divalent metal ions to show any enzymatic activity. Both of RGLs showed a substrate preference for RG-I compared to other pectin domains. Bo4416 and Bo3128 primarily yielded unsaturated RG oligosaccharides, with Bo3128 also producing them with short side chains, with yields of 32.4 and 62.4%, respectively. Characterization of both RGLs contribute to the preparation of rhamnogalacturonan oligosaccharides, as well as for the analysis of the fine structure of RG-I pectins.",

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author = "Weiyang Wang and Yibing Wang and Haoting Yi and Yang Liu and Guojing Zhang and Le Zhang and Mayo, {Kevin H.} and Ye Yuan and Yifa Zhou",

note = "Funding Information: National Natural Science Foundation of China (No. 32000907); Jilin Provincial Department of Science and Technology, Medicine and Health Project (No. 20200504005YY); and the Scientific and Technologic Foundation of Jilin Province (Nos. 20200201004JC and 20200708059YY). Funding Information: National Natural Science Foundation of China (No. 32000907); Jilin Provincial Department of Science and Technology, Medicine and Health Project (No. 20200504005YY); and the Scientific and Publisher Copyright: Copyright {\textcopyright} 2022 Wang, Wang, Yi, Liu, Zhang, Zhang, Mayo, Yuan and Zhou.",

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AU - Wang, Yibing

AU - Yi, Haoting

AU - Liu, Yang

AU - Zhang, Guojing

AU - Zhang, Le

AU - Mayo, Kevin H.

AU - Yuan, Ye

AU - Zhou, Yifa

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N2 - Rhamnogalacturonan lyase (RGL) cleaves backbone α-1,4 glycosidic bonds between L-rhamnose and D-galacturonic acid residues in type I rhamnogalacturonan (RG-I) by β-elimination to generate RG oligosaccharides with various degrees of polymerization. Here, we cloned, expressed, purified and biochemically characterized two RGLs (Bo3128 and Bo4416) in the PL11 family from Bacteroides ovatus ATCC 8483. Bo3128 and Bo4416 displayed maximal activity at pH 9.5 and pH 6.5, respectively. Whereas the activity of Bo3128 could be increased 1.5 fold in the presence of 5 mM Ca2+, Bo4416 required divalent metal ions to show any enzymatic activity. Both of RGLs showed a substrate preference for RG-I compared to other pectin domains. Bo4416 and Bo3128 primarily yielded unsaturated RG oligosaccharides, with Bo3128 also producing them with short side chains, with yields of 32.4 and 62.4%, respectively. Characterization of both RGLs contribute to the preparation of rhamnogalacturonan oligosaccharides, as well as for the analysis of the fine structure of RG-I pectins.

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KW - type I rhamnogalacturonan

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Biochemical Characterization of Two Rhamnogalacturonan Lyases From Bacteroides ovatus ATCC 8483 With Preference for RG-I Substrates

Abstract

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Keywords

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