1999 Fiscal Year Final Research Report Summary
Creation of stable enzymes for PCR by evolutionary molecular engineering
| Project/Area Number |
09558081
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
Structural biochemistry
|
|---|
| Research Institution | Tokyo University of Pharmacy & Life Science |
|---|
Principal Investigator |
OSHIMA Tairo School of Life Science, Tokyo University of Pharmacy & Life Science Professor, 生命科学部, 教授 (60167301)
|
|---|
| Project Period (FY) |
1997 – 1999
|
| Keywords | isopropylmalate dehydrogenase / tryptophan synthetase / nicking / structural unit / TIM barrel / fragmented enzyme |
|---|
| Research Abstract |
The final goal of the present project is to create stable enzymes and polymerases for PCR and the related techniques by laboratory evolution. For this purpose, the investigator devised two laboratory evolution systems ; (1) using an extreme thermophile, Thermus thermophilus as a host, genes coding for mesophile enzymes were integrated into the thermophile chromosome. (2) In the other system, E.coli is used as the host and genes coding for thermophile enzymes were inserted into the chromosomal DNA.When isopropylmalate dehydrogenase is used as a model enzyme, the dehydrogenases from many mesophiles including yeast were successfully stabilized. Likewise, the catalytic activity at room temperature of the Thermus thermophilus enzyme was improved by this method. At present stabilization of E.coli DNA polymerase is under progress.
|
