한국식품영양과학회지 (Journal of the Korean Society of Food Science and Nutrition)

  • 제43권9호
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  • Pages.1388-1393
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  • 2014
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  • 1226-3311(pISSN)
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  • 2288-5978(eISSN)
한국식품영양과학회 (The Korean Society of Food Science and Nutrition)
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Cyclodextrin Glucanotransferase를 이용한 아밀로펙틴 클러스터의 생산

Enzymatic Production of Amylopectin Cluster Using Cyclodextrin Glucanotransferase

  • 이혜원 (한림대학교 식품영양학과) ;
  • 전혜연 (한림대학교 식품영양학과) ;
  • 최혜정 (한림대학교 식품영양학과) ;
  • 심재훈 (한림대학교 식품영양학과)
  • 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)
  • 투고 : 2014.04.21
  • 심사 : 2014.06.05
  • 발행 : 2014.09.30
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초록

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.

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.

키워드

참고문헌

  1. Park JH, Kim HJ, Kim YH, Cha HJ, Kim YW, Kim TJ, Kim YR, Park KH. 2007. The action mode of Thermus aquaticus YT-1 4-$\alpha$-glucanotransferase and its chimeric enzymes introduced with starch-binding domain on amylose and amylopectin. Carbohydr Polym 67: 164-173. https://doi.org/10.1016/j.carbpol.2006.05.018
  2. Kim BK, Kim HI, Moon TW, Choi SJ. 2014. Branch chain elongation by amylosucrase: production of waxy corn starch with a slow digestion property. Food Chem 152: 113-120. https://doi.org/10.1016/j.foodchem.2013.11.145
  3. Myers AM, Morell MK, James MG, Ball SG. 2000. Recent progress toward understanding biosynthesis of the amylopectin crystal. Plant Physiol 122: 989-998. https://doi.org/10.1104/pp.122.4.989
  4. Grundy SM. 1998. Multifactorial causation of obesity: implications for prevention. Am J Clin Nutr 67: 563S-572S.
  5. Abbott WG, Swinburn B, Ruotolo G, Hara H, Patti L, Harper I, Grundy SM, Howard BV. 1990. Effect of a highcarbohydrate, low-saturated-fat diet on apolipoprotein B and triglyceride metabolism in Pima Indians. J Clin Invest 86:642-650. https://doi.org/10.1172/JCI114756
  6. Lee CK, Le QT, Kim YH, Shim JH, Lee SJ, Park JH, Lee KP, Song SH, Auh JH, Lee SJ, Park KH. 2008. Enzymatic synthesis and properties of highly branched rice starch amylose and amylopectin cluster. J Agric Food Chem 56: 126-131. https://doi.org/10.1021/jf072508s
  7. Seo NS, Roh SA, Auh JH, Park JH, Kim YR, Park KH. 2007. Structural characterization of rice starch in rice cake modified by Thermus scotoductus 4-$\alpha$-glucanotransferase (TS$\alpha$ GTase). J Food Sci 72: C331-C336. https://doi.org/10.1111/j.1750-3841.2007.00428.x
  8. Shim JH, Seo NS, Roh SA, Kim JW, Cha HJ, Park KH. 2007. Improved bread-baking process using Saccharomyces cerevisiae displayed with engineered cyclodextrin glucanotransferase. J Agric Food Chem 55: 4735-4740. https://doi.org/10.1021/jf070217d
  9. Shim JH, Kim YW, Kim TJ, Chae HY, Park JH, Cha HJ, Kim JW, Kim YR, Schaefer T, Spendler T, Moon TW, Park KH. 2004. Improvement of cyclodextrin glucanotransferase as an antistaling enzyme by error-prone PCR. Protein Eng Des Sel 17: 205-211. https://doi.org/10.1093/protein/gzh035
  10. Lee SH, Kim YW, Lee SY, Auh JH, Yoo SS, Kim TJ, Kim JW, Kim ST, Rho HJ, Choi JH, Kim YB, Park KH. 2002. Modulation of cyclizing activity and thermostability of cyclodextrin glucanotransferase and its application as an antistaling enzyme. J Agric Food Chem 50: 1411-1415. https://doi.org/10.1021/jf010928q
  11. Chung HJ, Yoon SH, Lee MJ, Kim MJ, Kweon KS, Lee IW, Kim JW, Oh BH, Lee HS, Spiridonova VA, Park KH. 1998. Characterization of a thermostable cyclodextrin glucanotransferase isolated from Bacillus stearothermophilus ET1. J Agric Food Chem 46: 952-959. https://doi.org/10.1021/jf970707d
  12. Bradford M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  13. Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685. https://doi.org/10.1038/227680a0
  14. Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination by reducing sugar. Anal Chem 31: 426-428. https://doi.org/10.1021/ac60147a030
  15. Yong HW, Kim SM, Shim JH. 2012. Enzymatic production of alkyl ${\beta}$-glucoside based on transglycosylation activity of celluclast. J Korean Soc Food Sci Nutr 41: 1417-1422. https://doi.org/10.3746/jkfn.2012.41.10.1417

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