Enzymatic hydrolysis of 1,3-1,4-β-glucosyl oligosaccharides by 1,3-1,4-β-glucanase from Synechocystis PCC6803: A comparison with assays using polymer and chromophoric oligosaccharide substrates. Fukamizo, T., Hayashi, K., Tamoi, M., Fujimura, Y., Kurotaki, H., Kulminskaya, A., & Kitaoka, M. Archives of Biochemistry and Biophysics, 478(2):187-194, 2008. cited By 8
Enzymatic hydrolysis of 1,3-1,4-β-glucosyl oligosaccharides by 1,3-1,4-β-glucanase from Synechocystis PCC6803: A comparison with assays using polymer and chromophoric oligosaccharide substrates [link]Paper  doi  abstract   bibtex   
The specificity of 1,3-1,4-β-glucanase from Synechocystis PCC6803 (SsGlc) was investigated using novel substrates 1,3-1,4-β-glucosyl oligosaccharides, in which 1,3- and 1,4-linkages are located in various arrangements. After the enzymatic reaction, the reaction products were separated and determined by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). As a result, SsGlc was found to hydrolyze the pentasaccharides, which possess three contiguous 1,4-β-glycosidic linkages (cellotetraose sequence) adjacent to 1,3-β-linkage, but none of the other oligosaccharides were hydrolyzed. To further analyze the specificity, kinetic measurements were performed using polymeric substrates and 4-methylumbelliferyl derivatives of laminaribiose and cellobiose (1,3-β-(Glc)2-MU and 1,4-β-(Glc)2-MU). The kcat/Km value obtained for barley β-glucan was considerably larger than that for lichenan, indicating that SsGlc prefers 1,3-1,4-β-glucan possessing a larger amount of cellotetraose sequence. This is consistent with the data obtained for 1,3-1,4-β-glucosyl oligosaccharides. However, the kcat/Km value obtained for 1,4-β-(Glc)2-MU was considerably lower than that for 1,3-β-(Glc)2-MU, suggesting inconsistency with the data obtained from the other natural substrates. It is likely that the kinetic data obtained from such chromophoric substrates do not always reflect the true enzymatic properties. © 2008 Elsevier Inc. All rights reserved.
@ARTICLE{Fukamizo2008187,
author={Fukamizo, T. and Hayashi, K. and Tamoi, M. and Fujimura, Y. and Kurotaki, H. and Kulminskaya, A. and Kitaoka, M.},
title={Enzymatic hydrolysis of 1,3-1,4-β-glucosyl oligosaccharides by 1,3-1,4-β-glucanase from Synechocystis PCC6803: A comparison with assays using polymer and chromophoric oligosaccharide substrates},
journal={Archives of Biochemistry and Biophysics},
year={2008},
volume={478},
number={2},
pages={187-194},
doi={10.1016/j.abb.2008.07.019},
note={cited By 8},
url={https://www.scopus.com/inward/record.uri?eid=2-s2.0-52249117218&doi=10.1016%2fj.abb.2008.07.019&partnerID=40&md5=55602366a3c9a982965d25b953ec29bc},
affiliation={Department of Advanced Bioscience, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan; Laboratory of Enzymology, Molecular and Radiation Biophysics Division, Petersburg Nuclear Physics Institute, Orlova roscha, 188300, Gatchina, Len. District, Russian Federation; National Food Research Institute, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8642, Japan},
abstract={The specificity of 1,3-1,4-β-glucanase from Synechocystis PCC6803 (SsGlc) was investigated using novel substrates 1,3-1,4-β-glucosyl oligosaccharides, in which 1,3- and 1,4-linkages are located in various arrangements. After the enzymatic reaction, the reaction products were separated and determined by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). As a result, SsGlc was found to hydrolyze the pentasaccharides, which possess three contiguous 1,4-β-glycosidic linkages (cellotetraose sequence) adjacent to 1,3-β-linkage, but none of the other oligosaccharides were hydrolyzed. To further analyze the specificity, kinetic measurements were performed using polymeric substrates and 4-methylumbelliferyl derivatives of laminaribiose and cellobiose (1,3-β-(Glc)2-MU and 1,4-β-(Glc)2-MU). The kcat/Km value obtained for barley β-glucan was considerably larger than that for lichenan, indicating that SsGlc prefers 1,3-1,4-β-glucan possessing a larger amount of cellotetraose sequence. This is consistent with the data obtained for 1,3-1,4-β-glucosyl oligosaccharides. However, the kcat/Km value obtained for 1,4-β-(Glc)2-MU was considerably lower than that for 1,3-β-(Glc)2-MU, suggesting inconsistency with the data obtained from the other natural substrates. It is likely that the kinetic data obtained from such chromophoric substrates do not always reflect the true enzymatic properties. © 2008 Elsevier Inc. All rights reserved.},
author_keywords={1,3-1,4-β-glucanase;  Cleavage specificity;  Cyanobacterium;  Glucosyl oligosaccharides;  HPAEC-PAD},
}
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