| Research Abstract |
The surface structure of porous glasses, which would be used as supports in DNA syntheses, was investigated. Silanol groups on the surface were estimated to be 7-9mumol・m^<-2>. The value was compared with the crystal structure of cristobalite, as a model structure of SiO_2 glass. Amino-alkyl functionalities obtained by silane coupling agents was estimated to be 2-3mumol・m^<-2>. Control of the surface densities of amino-alkyl functionalities by adding alkyl silane, such as methyl trimethoxysilane, was useful to avoid the stereochemical interactions hindering the growth of the DNA chains. Amino-alkylation of the glass surface of 5mumol・m^<-2> was found as the optimum density. Pore diameters and chemical compositions of the skeletal glass were optimized.
A clone of carboxymethyl cellulose degrading enzyme obtained from a wild strain of Bacillus was established using E. coli K12, HB101, as a host, and pBR322 plasmid as a vector. This could be used in further studies to develop wider range of uses of the porous glass substrata, in many aspects of protein engineering, such as DNA and peptide syntheses, immobilized enzymes, affinity chromatography, etc.
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