| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2006: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
1) A novel biphenyl dioxygenase BphA(2-2) was produced, and tested its substrate specificity. BphA(2-2) + BphB converted 7-hydroxyflavone and chrysin (5,7-dihydroxyflavone) to the correspondiong 1,2-diols (7,2',3'-hydroxyflavone and 5,7,2',3'-hydroxyflavone, novel compounds). The novel flavones showed potent cell protective (=antioxidative) activity against N18-RE-105 cells.
2) Novel P450s (pfusion and Balk) was obtained from marine bacteria. The substrate specifities of these P450s was studied using the aromatic compounds. P450 (pfusion) inserted phenolic OH to the aromatic rings. P450 (Balk) mainly converted the allylic methyls to the corresponding primary alchols. But, in the case of 2-bromo-phenol, P450 (Balk) inserted phenolic OH to the benzene ring.
3) A sequential bioconversion using Bphs (BphA + BphB) and geranyl transferase (NphB) were tested. The geranyl transferase (NphB) was presented by Dr. Kuzuyama (The university of Tokyo).
A : At first, chrysin was converted to the corresponding diols (B ring) using BphA (2-2) + BphB. Next, the converted product was converted with NPhB. As the result, 3 geranyled products were obtained. The structure of the major product was analysed by NMR and MS, and determined to be 5'-monogeranylated compounds. The structures of the others were under investigation, and proposed to be di-geranylated ones (5' and X). These geranylated flavones were all novel compounds and showed potent antioxidative activites in the N18-RE-105 assay.
B : At first, chrysin was converted using NphB. 6-C-geranyl chrysin and 7-O-geranyl chrysin were obtained as the converted product. These products were tested for the bioconversion by BphA (2-2) + BphB, but no product was obtained.
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