Mutation strategies for obtaining chitooligosaccharides with longer chains by transglycosylation reaction of family GH18 chitinase
| Autor: | Tamo Fukamizo, Shoko Shinya, Yuka Kanda, Takayuki Ohnuma, Natchanok Sritho, Wipa Suginta, Paknisa Sirimontree |
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| Rok vydání: | 2014 |
| Předmět: |
Glycosylation
Mutant Gene Expression Oligosaccharides Protein Engineering Applied Microbiology and Biotechnology Biochemistry Acetylglucosamine Analytical Chemistry Catalysis Hydrolysis Bacterial Proteins Glycoside hydrolase Site-directed mutagenesis Molecular Biology Incubation Vibrio Aspartic Acid Alanine biology Vibrio harveyi Chitinases Organic Chemistry General Medicine biology.organism_classification Molecular Docking Simulation Mutation Chitinase Mutagenesis Site-Directed biology.protein Asparagine Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 78:2014-2021 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1080/09168451.2014.948373 |
| Popis: | Enhancing the transglycosylation (TG) activity of glycoside hydrolases does not always result in the production of oligosaccharides with longer chains, because the TG products are often decomposed into shorter oligosaccharides. Here, we investigated the mutation strategies for obtaining chitooligosaccharides with longer chains by means of TG reaction catalyzed by family GH18 chitinase A from Vibrio harveyi (VhChiA). HPLC analysis of the TG products from incubation of chitooligosaccharide substrates, GlcNAcn, with several mutant VhChiAs suggested that mutant W570G (mutation of Trp570 to Gly) and mutant D392N (mutation of Asp392 to Asn) significantly enhanced TG activity, but the TG products were immediately hydrolyzed into shorter GlcNAcn. On the other hand, the TG products obtained from mutants D313A and D313N (mutations of Asp313 to Ala and Asn, respectively) were not further hydrolyzed, leading to the accumulation of oligosaccharides with longer chains. The data obtained from the mutant VhChiAs suggested that mutations of Asp313, the middle aspartic acid residue of the DxDxE catalytic motif, to Ala and Asn are most effective for obtaining chitooligosaccharides with longer chains. |
| Databáze: | OpenAIRE |
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