Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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High Yield Production of Cyclofructan by Deletion Mutant Enzyme of Cycloinulooligosaccharide Fructanotransferase

Cycloinulooligosaccharide fructanotransferase의 결손변이효소에 의한 cyclofructan의 고효율 생산

  • Park Jung-Ha (Department of Life Science and Biotechnology, College of Natural Science, Dongeui University) ;
  • Kwon Hyun-Ju (Department of Life Science and Biotechnology, College of Natural Science, Dongeui University) ;
  • Kim Byung-Woo (Department of Life Science and Biotechnology, College of Natural Science, Dongeui University)
  • 박정하 (동의대학교 생명응용과학과) ;
  • 권현주 (동의대학교 생명응용과학과) ;
  • 김병우 (동의대학교 생명응용과학과)
  • Published : 2006.02.01
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Abstract

This study investigated the optimal conditions of high yield production of cyclofructan (CF) using recombinant deletion mutant enzyme CFT108 which is constructed by N-terminal deletion from cycloinulooligosaccharide fructanotransferase (CFTase) gene of Penibacillus polymyxa. The production yield was dependent on reaction time, substrate concentration and enzyme concentration. The optimum reaction time for industrial purpose was achieved at 3 hr reaction. The optimal concentrations of substrate and enzyme were found to be $2\%$ inulin and 40 unit/ g inulin, respectively. At optimum condition, 9.5 g/l of maximum yield and $47.5\%$ of conversion efficacy were achieved. For purification of CF produced, the reaction mixture was treated with 1 unit/ml exoinulinase and then added $3\%$ CaO three times with blowing $CO_2$ gas, resulted in $95\%$ purity.

본 연구는 Penibacillus polymyxa 균주의 CFTase 유전자를 결손 변이시킨 고효율 효소 CFT108을 이용하여 cyclofructan (CF)의 대량생산 조건을 검토하고 생산된 CF의 순수 분리 정제 공정을 개발하였다. CF의 대량생산 조건은 $2\%$의 inulin 기질과 40 unit/g inulin의 기질에서 3시간 반응시켰을 때 최대 생산량 9.5 g/l의 수율을 달성할 수 있었으며, 이때의 in-ulin의 CF 전환율은 $47.5\%$였다. 생산된 CF를 순수 분리 정제하기 위해서 CFTase 반응액 을 exoinulinase 1 unit/ml로 6시간 처리하여 미 반응 inulin을 단당화시켰으며, 유리된 fructose는 $3\%$ CaO로 $CO_2$가스 하에서 $80^{\circ}C$, 10분간 3회 흡착제거 시킴으로써 순수 정제하였다. 정제된 CF를 HPLC로 확인한 결과 정제도는 $95\%$ 이상이였으며, $CF_6,\;CF_7,\;CF_8$ 비율이 72 : 27 : 1 이였다.

Keywords

References

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