1993 Fiscal Year Final Research Report Summary
cDNA cloning of enzymes involved in the synthesis and degradation of cell wall-matrix polysaccharides
| Project/Area Number |
03640574
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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 |
植物生理学
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| Research Institution | OSAKA CITY UNIVERSITY |
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Principal Investigator |
KAMISAKA Seiichiro Osaka City Univ. Faculty of Science, Professor, 理学部, 教授 (60047214)
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| Co-Investigator(Kenkyū-buntansha) |
HOSON Takayuki Osaka City Univ. Associate Professor, 理学部, 助教授 (70135771)
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| Project Period (FY) |
1991 – 1993
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| Keywords | Auxin / Xyloglucan / Glucanase / Fucosidase / Arabinoxylan / Xyloglucanase / Feruloy1-CoA / Esterase |
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| Research Abstract |
The mechanical properties of cell walls are determined by cell wallconstituting molecules, particularly matrix polysaccharides. The goal of this study is to clone cDNA of enzymes involved in the synthesis and degradation of xyloglucans and arabinoxylans, which are responsible for determining cell wall extensibility. Results are as follows :
(1) Matrix polysaccharide-degrading enzymes : Xyloglucan degrading enzymes were solubilized with NaCl from Vigna epicotyl cell walls. The enzymes were fractionated and purified using Con A-Sepharose, Mono S, Superdex 75, and Phenyl Superose column. The enzymes are separated into 3 peaks on Mono S colum. alpha-Fucosidases which eliminate fucos-sede chain of xyloglucans were separated into Mono S-binding and -not binding fractions.
(2) Matrix polysaccharide-modifying enzymes : Using Golgi-vesicle fraction, the screening of the enzyme that ester-links ferulic acid to arabinose moieties of arabinoxylans. Oat arabinoxylans and synthesitic feruloyl-CoA were used as the substrates. Unfortunately, the enzyme activity was not detected, because of the high activity of feruloyl-CoA esterase in the enzyme preparation. At present, extensive effort has been done to eliminate the esterase from the enzyme preparation.
The present srudy will soon lead to the cloning of cDNA of the enzymes responsible for the synthesis and degradation of matrix polysaccharides determing the mechanical properteis of cell walls, although the cDNA cloning was not done within the research period of 3 years.
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