Biosynthesis of New Oligosaccharides via Acceptor Reaction using Fructosyloligosaccharide as an Acceptor

Fructosyloligosaccharide를 Acceptor 반응의 기질로 사용한 새로운 올리고당의 생합성

  • Published : 1999.06.01

Abstract

Acceplor reaction of glucosyltr~ansferase of Streptococcus ,SO~~-~IZLIS with f ~ ~ t o ~ y l o l i g o ~ a ~ ~ h a r i d e ~ was studied for the biosynthesis of novel olgisaccharides. First, bacilracin resistant mutants were selected by mutagenesis of Streptococcus sobrimis ATCC27351. Among these mutants 4 strains were selected by resistance to bacitracin and increase of glucosyltransferase. Acceptor reaction of maltose was analyzed by TLC and image analysis. There were differences in the specificity of the acceptor reaclion by Ule glucosylumsferase between mother strain (Streptococcus sobrinus ATCC2735) and bacitracin resistant mutants (Streptococcus sobrinus BR24C, Strepfococcus sobrinus CH-5). Molher strain did ilot show an acceptor reaction with fructosyloligosaccharides such as 1-keqtose and nystose. Acceptor reaction products of turailose and 1-kestose with glucosyltransferase (GW-S) of Streptococcus sobrini~s BR24C were TEX>\6^{3}$-$\alpha$-D-glucopyranosyl \3^{2}$-O-$\alpha$-D-fructose (glucose-fructose-glucose) and \6^{4}$-$\alpha$-D-glucopyranosyl \1^{3}$-$\beta$-D-~-h~ctofuranos~~ sucrose (glucose-glucosefructose- fructose). respectively These are novel oligosaccharides which can be produced only by enzymatic reaction.

충치 원인균의 일종인 Streptococcus sobrinus ATCC27351 균주를 자외선 조사와 NTG 처리에 의하여 bacitracin 내성변이주를 얻었다. 선발된 변이주들 중 glucosyltransferase 활성이 모균주보다 증가된 변이주 4균주를 선발하였다. 이들의 bacitracin 내성은 모균주 보다 5~48배, glucosyltransferase 활성은 약 2배 증가하였다. 모균주의 glucosyltransferase를 이용하여 maltose acceptor 반응을 하여 \ulcorner 와 image analysis 로 분석하였다. 그 결과 sucrose 와 acceptor를 사용하여 반응을 시킨 결과 모균주와 변이주의 glucose 전이효소 사이에는 acceptor 특이성에 차이가 있었다. 모균주의 효소와는 반응하지 않는 acceptor 인 fructose를 함유하는 1-kestose와 nystose 그리고 turanose를 acceptor 로 하여 변이주 S. sobrinus BR24C 균주의 glucosyltransferase와 반응을 시켰다. 얻어진 acceptor 산물을 정제하여 \sup 1\H, \sup 13\C-NMR 분석을 한 결과, 자연계에서는 발견되지 않는 새로운 구조의 올리고당 \6^{3}$-$\alpha$-D-glucopyranosyl \3^{2}$-O-$\alpha$-D-glucopyranosyl-D-fructose와 \6^{4}$-$\alpha$-D-glucopyranosyl \1^{3}$-$\beta$-D-fructofuranosyl sucrose을 확인하였다.

Keywords

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