Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family
Glucosylsucroses are potentially useful as additives in cosmetic and pharmaceutical formulations. Although enzymatic synthesis of glucosylsucroses is the most efficient method for their production, the key enzyme that produces them has remained unknown. Here, we report that glucosylsucrose synthase...
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| Language: | English |
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China Science Publishing & Media Ltd.
2022-04-01
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| Series: | Acta Biochimica et Biophysica Sinica |
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| Online Access: | https://www.sciengine.com/doi/10.3724/abbs.2022034 |
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| author | Han Qiuyu Yao Yuan Liu Yuhan Zhang Wenlu Yu Jinyi Na Heya Liu Tianhao Mayo Kevin H. Su Jiyong |
| author_facet | Han Qiuyu Yao Yuan Liu Yuhan Zhang Wenlu Yu Jinyi Na Heya Liu Tianhao Mayo Kevin H. Su Jiyong |
| author_sort | Han Qiuyu |
| collection | DOAJ |
| description | Glucosylsucroses are potentially useful as additives in cosmetic and pharmaceutical formulations. Although enzymatic synthesis of glucosylsucroses is the most efficient method for their production, the key enzyme that produces them has remained unknown. Here, we report that glucosylsucrose synthase from Thermosynechococcus elongatus (TeGSS) catalyzes the synthesis of glucosylsucrose using sucrose and UDP-glucose as substrates. These saccharides are homologous to glucosylsucroses produced by Nostoc sp. PCC 7120 (referred to as protein alr1000). When the ratio of UDP-glucose to sucrose is relatively high, TeGSS from cyanobacteria can hydrolyze excess UDP-glucose to UDP and glucose, indicating that sucrose provides a feedback mechanism for the control of glucosylsucrose synthesis. In the present study, we solved the crystal structure of TeGSS bound to UDP and sucrose. Our structure shows that the catalytic site contains a circular region that may allow glucosylsucroses with a right-hand helical structure to enter the catalytic site. Because active site residues Tyr18 and Arg179 are proximal to UDP and sucrose, we mutate these residues (i.e., Y18F and R179A) and show that they exhibit very low activity, supporting their role as catalytic groups. Overall, our study provides insight into the catalytic mechanism of TeGSS. |
| format | Article |
| id | doaj-art-cdc729e43dfe47739c06f7bc2bf46d38 |
| institution | OA Journals |
| issn | 1672-9145 |
| language | English |
| publishDate | 2022-04-01 |
| publisher | China Science Publishing & Media Ltd. |
| record_format | Article |
| series | Acta Biochimica et Biophysica Sinica |
| spelling | doaj-art-cdc729e43dfe47739c06f7bc2bf46d382025-08-20T01:52:15ZengChina Science Publishing & Media Ltd.Acta Biochimica et Biophysica Sinica1672-91452022-04-015453754710.3724/abbs.202203420d259ccStructural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase familyHan Qiuyu0Yao Yuan1Liu Yuhan2Zhang Wenlu3Yu Jinyi4Na Heya5Liu Tianhao6Mayo Kevin H.7Su Jiyong8["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]["Media Academy, Jilin Engineering Normal University, Changchun 130052, China"]["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]["Department of Biochemistry, Molecular Biology & Biophysics, University of Minnesota, Minneapolis, MN 55455, USA"]["Engineering Research Center of Glycoconjugates Ministry of Education, Jilin Provincial Key Laboratory of Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun 130024, China"]Glucosylsucroses are potentially useful as additives in cosmetic and pharmaceutical formulations. Although enzymatic synthesis of glucosylsucroses is the most efficient method for their production, the key enzyme that produces them has remained unknown. Here, we report that glucosylsucrose synthase from Thermosynechococcus elongatus (TeGSS) catalyzes the synthesis of glucosylsucrose using sucrose and UDP-glucose as substrates. These saccharides are homologous to glucosylsucroses produced by Nostoc sp. PCC 7120 (referred to as protein alr1000). When the ratio of UDP-glucose to sucrose is relatively high, TeGSS from cyanobacteria can hydrolyze excess UDP-glucose to UDP and glucose, indicating that sucrose provides a feedback mechanism for the control of glucosylsucrose synthesis. In the present study, we solved the crystal structure of TeGSS bound to UDP and sucrose. Our structure shows that the catalytic site contains a circular region that may allow glucosylsucroses with a right-hand helical structure to enter the catalytic site. Because active site residues Tyr18 and Arg179 are proximal to UDP and sucrose, we mutate these residues (i.e., Y18F and R179A) and show that they exhibit very low activity, supporting their role as catalytic groups. Overall, our study provides insight into the catalytic mechanism of TeGSS.https://www.sciengine.com/doi/10.3724/abbs.2022034catalytic mechanismcrystal structureglucosylsucroseUDP-glucoseα-1,2-glucosyltransferase |
| spellingShingle | Han Qiuyu Yao Yuan Liu Yuhan Zhang Wenlu Yu Jinyi Na Heya Liu Tianhao Mayo Kevin H. Su Jiyong Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family Acta Biochimica et Biophysica Sinica catalytic mechanism crystal structure glucosylsucrose UDP-glucose α-1,2-glucosyltransferase |
| title | Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family |
| title_full | Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family |
| title_fullStr | Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family |
| title_full_unstemmed | Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family |
| title_short | Structural basis for glucosylsucrose synthesis by a member of the α-1,2-glucosyltransferase family |
| title_sort | structural basis for glucosylsucrose synthesis by a member of the α 1 2 glucosyltransferase family |
| topic | catalytic mechanism crystal structure glucosylsucrose UDP-glucose α-1,2-glucosyltransferase |
| url | https://www.sciengine.com/doi/10.3724/abbs.2022034 |
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