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Enzymatic Production of Amylopectin Cluster Using Cyclodextrin Glucanotransferase

Cyclodextrin Glucanotransferase를 이용한 아밀로펙틴 클러스터의 생산

  • Lee, Hye-Won (Department of Food Science and Nutrition, Hallym University) ;
  • Jeon, Hye-Yeon (Department of Food Science and Nutrition, Hallym University) ;
  • Choi, Hyejeong (Department of Food Science and Nutrition, Hallym University) ;
  • Shim, Jae-Hoon (Department of Food Science and Nutrition, Hallym University)
  • 이혜원 (한림대학교 식품영양학과) ;
  • 전혜연 (한림대학교 식품영양학과) ;
  • 최혜정 (한림대학교 식품영양학과) ;
  • 심재훈 (한림대학교 식품영양학과)
  • Received : 2014.04.21
  • Accepted : 2014.06.05
  • Published : 2014.09.30

Abstract

To enzymatically prepare amylopectin cluster (APC), cyclodextrin glucanotransferase (CGTase I-5) and its mutant enzyme from alkalophilic Bacillus sp. I-5 were employed, after which the hydrolysis patterns of CGTase wild-type and its mutant enzyme toward amylopectin were investigated using multi-angle laser light scattering. CGTase wild-type dramatically reduced the molecular weight of waxy rice starch at the initial reaction, whereas the mutant enzyme degraded waxy rice starch relatively slowly. Based on the results, the molecular weight of one cluster of amylopectin could be about $10^4{\sim}10^5g/mol$. To determine production of cyclic glucans from amylopectin, matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed. CGTase I-5 produced various types of cyclic maltooligosaccharides from amylopectin, whereas the mutant enzyme hardly produced any.

선행연구에서 얻은 alkalophilic Bacillus I-5 유래의 CGTase wild-type과 가수분해능이 강화된 mutant 효소를 활용하여 waxy rice starch로부터 아밀로펙틴 클러스터를 제조하였다. SEC-MALLS-RI 분석법으로 CGTase wild-type과 mutant 효소가 처리된 시료의 평균 분자량을 확인한 결과 10분가량의 효소반응으로 두 반응 모두 평균 분자량은 $10^4{\sim}10^5Da$으로 급격히 감소하였음을 확인하였으며, 일정 반응 시간이 경과한 이후에는 더 이상 분자량의 감소가 일어나지 않음으로 미루어 시료가 아밀로펙틴 클러스터 단위로 분해되었으며 그 분자량은 $10^4{\sim}10^5Da$ 정도임을 알 수 있다. 또한 MALDI-TOF/MS 분석을 통하여 CGTase wild-type은 다양한 종류의 cyclic 형태의 maltodextrin을 생성하고 있으며 mutant 효소는 주로 소량의 maltooligosaccharide 들을 생산함을 확인하였다.

Keywords

Acknowledgement

Supported by : 중소기업청

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