Kinetic relationships with processivity in Serratia marcescens family 18 glycoside hydrolases |
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| Institution: | 1. School of Biological Sciences, Department of Molecular & Biomedical Science, Research Centre for Infectious Diseases, University of Adelaide, Adelaide, 5005, Australia;2. Institute for Glycomics, Griffith University Gold Coast Campus, Queensland, 4222, Australia;1. Department of Food and Nutritional Sciences, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan;2. Fuji Oil Holdings Inc., Daibiru Honkan Building, 3-6-32, Nakanoshima, Kita-ku, Osaka-shi, Osaka, 530-0005, Japan;1. Department of Orthopaedics, Gongli Hospital of Pudong New Area, Shanghai, China;2. Ningxia Medical University, Yinchuan, China;1. University of Chinese Academy of Sciences, Beijing, 100049, China;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China;3. Institute of Biological Chemistry, Academia Sinica, Taipei, 11529, Taiwan;4. State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, Wuhan, 430062, China;1. Graduate School of Technology, Industrial and Social Science, Tokushima University, Tokushima, 770-8506, Japan;2. Division of Education for Global Standard, Institute of Liberal Arts and Science, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan;3. Central Research Resource Branch, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan;4. Division of Functional Genomics, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan |
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| Abstract: | In nature, recalcitrant polysaccharides such as chitin and cellulose are degraded by glycoside hydrolases (GH) that act synergistically through different modes of action including attack from reducing-end and nonreducing-end (exo-mode) and random (endo-mode) on single polysaccharide chains. Both modes can be combined with a processive mechanism where the GH remain bound to the polysaccharide to perform multiple catalytic steps before dissociation into the solution. In this work, we have determined association rate constants and their activation paramaters for three co-evolved GHs from Serratia marcescens (SmChiA, SmChiB, and SmChiC) with an oligomeric substrate. Interestingly, we observe a positive correlation between the association rate constants and processive ability for the GHs. Previously, a positive correlation has been observed between substrate binding affinity and processive ability. SmChiA with highest processive ability of the three GHs bind with a kon of 11.5 ± 0.2 μM?1s?1, which is five-fold and 130-fold faster than SmChiB (less processive) and SmChiC (nonprocessive), respectively. |
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| Keywords: | Glycoside hydrolase Processivity Kinetics Thermodynamics |
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