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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제18권 제6호
- 발행연도
- 2008.1
- 수록면
- 1,064 - 1,069 (6page)
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초록· 키워드
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Levan fructotransferase (LFTase) preferentially catalyzes the transfructosylation reaction in addition to levan hydrolysis, whereas other levan-degrading enzymes hydrolyze levan into a levan-oligosaccharide and fructose. Based on sequence comparisons and enzymatic properties, the fructosyl transfer activity of LFTase is proposed to have evolved from levanase. In order to probe the residues that are critical to the intramolecular fructosyl transfer reaction of the Microbacterium sp. AL-210 LFTase, an error-prone PCR mutagenesis process was carried out, and the mutants that led to a shift in activity from transfructosylation towards hydrolysis of levan were screened by the DNS method. After two rounds of mutagenesis, TLC and HPLC analyses of the reaction products by the selected mutants revealed two major products; one is a di-D-fructose- 2,6':6,2'-dianhydride (DFAIV) and the other is a levanbiose.
The newly detected levanbiose corresponds to the reaction product from LFTase lacking transferring activity. Two mutants (2-F8 and 2-G9) showed a high yield of levanbiose (38-40%) compared with the wild-type enzyme, and thus behaved as levanases. Sequence analysis of the individual mutants responsible for the enhanced hydrolytic activity indicated that Asn-85 was highly involved in the transfructosylation activity of LFTase.
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참고문헌 (21)
참고문헌 신청
Alberto, F. / 2004 / Crystal structure of inactivated Thermotoga maritima invertase in complex with the trisaccharide substrate raffinose / Biochem 395 : 457 ~ 462
Avigad, G. / 1963 / Cleavage of fructans to levanbiose by a specific hydrolase. Bull. Res. Counc / Isr 11A4 : 253 ~ 257
Avigad, G. / 1996 / Fructan hydrolases / Methods Enzymol 8 : 621 ~ 628
Cairns, A. J. / 2003 / Fructan biosynthesis in transgenic plants. J. Exp / Bot 54 : 549 ~ 567
Cha, J. / 2001 / Molecular and enzymatic characterization of a levan fructotransferase from Microbacterium sp. AL-210. J / Biotechnol 91 : 49 ~ 61
Avigad, G. / 1963 / Cleavage of fructans to levanbiose by a specific hydrolase. Bull. Res. Counc / Isr 11A4 : 253 ~ 257
Avigad, G. / 1996 / Fructan hydrolases / Methods Enzymol 8 : 621 ~ 628
Cairns, A. J. / 2003 / Fructan biosynthesis in transgenic plants. J. Exp / Bot 54 : 549 ~ 567
Cha, J. / 2001 / Molecular and enzymatic characterization of a levan fructotransferase from Microbacterium sp. AL-210. J / Biotechnol 91 : 49 ~ 61
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