1987 Fiscal Year Final Research Report Summary
Production of Useful Oligosaccharides by Lytic Enzymes
| Project/Area Number |
61560138
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
製造化学・食品
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| Research Institution | Department of Agricultural Chemistry. Faculty of Agriculture Shizuoka University |
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Principal Investigator |
USUI Taichi Faculty of Agriculture, Shizuoka University, 農学部, 助教授 (50111802)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIDO Yoshiharu Faculty of Science, Tokyo Institute of Technology, 理学部, 教授 (60016037)
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| Project Period (FY) |
1986 – 1987
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| Keywords | Lytic enzyme / Lysozyme / Chitinase / Useful oligosaccharide by enzymatic synthesis / 糖転移反応 |
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| Research Abstract |
Chitinase from the culture filtrates of Nocardia orientalis IFO 12806 or Trichoderma reesei was purified to apparent homogeneity by successive column chromatographies. The enzyme, which is essentially a hydrolase, also catalyzed a transglycosylation reaction on tetra-N-acetyl chitotetraose(GlcNAc)_4 and penta N-acetyl-chitopentaose(GLcNAc)_5. The enzyme converted the tetrasaccharide into hexa-N-acetyl-chitohexaose(GlcNAc)_6 and di-N-acetyl-chitobiose(GLcNAc)_2 as the major products. The corresponding values for penta-N-acetyl-chitopentaose(GLcNAc)_5 were hepta-N-acetyl-chitoheptaose(GLcNAc)_3. The rate of the transglycosylation depended on the temperature, the concentration of substrate and the pH.
Transchitooligosylation from (GlcNAc)_5 to the 4-position of p-nitrophenyl 2-acetamido-2-deoxy-<beta>-D-glucopyranoside(PNP-GlcNAc) was efficiently induced through lysozyme catalysis in an aqueous solution containing methanol with a high concentration. Use of the aqueous methanol system in this reaction not only guaranteed solubility of PNP-GlcNAc substrate, but also resulted in a remarkable increase in p-nitrophenyl-penta-N-acetyl-<beta>-chitopentaoside(PNP-(GlcNAc)_5) production. PNP-(GlcNAc)_5 was substrate for lysozyme assay compared with p-nitrophenyl-tetra-N-acetyl-<beta>-chitotetraoside (PNP-(GlcNAc)_4).
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