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Substrate Specifici...
Substrate Specificities of Variants of Barley (1,3)- and (1,3;1,4)-β-d-Glucanases Resulting from Mutagenesis and Segment Hybridization
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- Kao, Mu-Rong (author)
- KTH,Albanova VinnExcellence Center for Protein Technology, ProNova,Glykovetenskap,School of Pharmacy, College of Pharmacy, Taipei Medical University, 250 Wuxing Street, Taipei 11031, Taiwan
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- Parker, Jake (author)
- School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Glen Osmond SA 5064, Australia;IBM Research Collaboratory for Life Sciences, Melbourne, Victoria 3010, Australia
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- Oehme, Daniel (author)
- IBM Research Collaboratory for Life Sciences, Melbourne, Victoria 3010, Australia
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- Chang, Shu-Chieh (author)
- KTH,Glykovetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Cheng, Lin-Chen (author)
- School of Pharmacy, College of Pharmacy, Taipei Medical University, 250 Wuxing Street, Taipei 11031, Taiwan
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- Wang, Damao (author)
- KTH,Glykovetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova,College of Food Science, Southwest University, Chongqing 400715, China
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- Srivastava, Vaibhav (author)
- KTH,Glykovetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Wagner, John M. (author)
- IBM Research Collaboratory for Life Sciences, Melbourne, Victoria 3010, Australia
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- Harris, Philip J. (author)
- School of Biological Sciences, The University of Auckland, Auckland Mail Centre, Private Bag 92019, Auckland 1142, New Zealand
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- Hsieh, Yves S. Y. (author)
- KTH,Glykovetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova,School of Pharmacy, College of Pharmacy, Taipei Medical University, 250 Wuxing Street, Taipei 11031, Taiwan
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(creator_code:org_t)
- American Chemical Society (ACS), 2024
- 2024
- English.
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In: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 63:9, s. 1194-1205
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Abstract
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- Barley (1,3;1,4)-β-d-glucanase is believed to have evolved from an ancestral monocotyledon (1,3)-β-d-glucanase, enabling the hydrolysis of (1,3;1,4)-β-d-glucans in the cell walls of leaves and germinating grains. In the present study, we investigated the substrate specificities of variants of the barley enzymes (1,3;1,4)-β-d-glucan endohydrolase [(1,3;1,4)-β-d-glucanase] isoenzyme EII (HvEII) and (1,3)-β-d-glucan endohydrolase [(1,3)-β-d-glucanase] isoenzyme GII (HvGII) obtained by protein segment hybridization and site-directed mutagenesis. Using protein segment hybridization, we obtained three variants of HvEII in which the substrate specificity was that of a (1,3)-β-d-glucanase and one variant that hydrolyzed both (1,3)-β-d-glucans and (1,3;1,4)-β-d-glucans; the wild-type enzyme hydrolyzed only (1,3;1,4)-β-d-glucans. Using substitutions of specific amino acid residues, we obtained one variant of HvEII that hydrolyzed both substrates. However, neither protein segment hybridization nor substitutions of specific amino acid residues gave variants of HvGII that could hydrolyze (1,3;1,4)-β-d-glucans; the wild-type enzyme hydrolyzed only (1,3)-β-d-glucans. Other HvEII and HvGII variants showed changes in specific activity and their ability to degrade the (1,3;1,4)-β-d-glucans or (1,3)-β-d-glucans to larger oligosaccharides. We also used molecular dynamics simulations to identify amino-acid residues or structural regions of wild-type HvEII and HvGII that interact with (1,3;1,4)-β-d-glucans and (1,3)-β-d-glucans, respectively, and may be responsible for the substrate specificities of the two enzymes.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Kao, Mu-Rong
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Parker, Jake
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Oehme, Daniel
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Chang, Shu-Chieh
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Cheng, Lin-Chen
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Wang, Damao
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Srivastava, Vaib ...
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Wagner, John M.
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Harris, Philip J ...
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Hsieh, Yves S. Y ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Biochemistry
- By the university
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Royal Institute of Technology