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Biological activity and analysis of α-glucosidase inhibitor from mulberry (Morus alba L.) wine

오디와인의 생리활성 및 α-glucosidase 저해제의 분석

  • Son, Woo-Rim (Technical Convergence Team, Biohelath Convergence Center of Daegu Techno-Park) ;
  • Choi, Sang-Won (Department of Food Science and Nutrition, Catholic University of Daegu)
  • 손우림 (대구테크노파크바이오헬스융합센터, 기술융합팀) ;
  • 최상원 (대구가톨릭대학교 식품영양학과)
  • Received : 2013.09.17
  • Accepted : 2013.12.17
  • Published : 2013.12.30

Abstract

Wine extracts of four different berry fruits, such as mulberry, blueberry, strawberry, and raspberry, were investigated for antioxidant, anti-tyrosinase, and ${\alpha}$-glucosidase activities by using in vitro assays. Additionally, quantitative changes of ${\alpha}$-glucosidase inhibitor in mulberry wine were determined by HPLC according to mulberry cultivars and fermentation process. Among four berry wines examined, mulberry wine showed the most potent ${\alpha}$-glucosidase inhibitory activity with 69.37% at 0.23 mg/mL, while blueberry and strawberry wines exhibited the strongest inhibition against DPPH radical and tyrosinase activity, respectively. Four compounds were isolated and purified from mulberry wine by a series of isolation procedures, such as solvent fractionation, and Diaion HP-20, ODS-A, and Sephadex LH-20 column chromatographies. Among them, Comp. 4 exerted the strongest ${\alpha}$-glucosidase inhibitory activity ($I_C_{50}=31.57{\mu}M$), and its chemical structure was identified as quercetin by UV and NMR spectral analysis. Finally, the "Daeseongppong" (16.83 ppm) muberry wine had larger amount of quercetin than the "Iksuppong" (14.85 ppm) and "Cheongilppong" (8.92 ppm) mulberry wines, but their contents of three mulberry wines decreased considerably with aging process. These results suggest that mulberry wine containing quercetin acted as ${\alpha}$-glucosidase inhibitor may be useful as a potential functional wine for improving diabetic disorder.

본 연구는 오디를 이용한 고부가가치의 기능성와인을 개발하기 위한 연구의 일환으로 먼저 4가지 베리류과실(오디, 블루베리, 딸기, 복분자)와인의 항당뇨, 항노화 및 항산화활성을 ${\alpha}$-glucosidase, tyrosinase 및 DPPH radical를 사용하여 측정한 후 ${\alpha}$-glucosidase 저해활성이 가장 강한 오디와인으로부터 저해제를 분리 및 동정하였다. 또한, 3 가지 뽕나무 품종(익수뽕, 대성뽕, 청일뽕) 오디를 이용하여 오디와인을 제조하여 품종과 발효에 따른 ${\alpha}$-glucosidase 저해제의 함량 변화를 측정한 결과는 다음과 같다. 4가지 베리류과실 와인의 항산화, 항당뇨 및 항노화 활성을 측정한 결과, 오디와인이 가장 높은 ${\alpha}$-glucosidase 저해활성을(69.37%) 나타내었으며, DPPH 라디컬 소거능은 블루베리(64.57%) 및 오디와인(62.48%)이 유사하게 높았으며, 딸기와인이 가장 높은 tyrosinase 저해활성(59.63%)을 나타내었다. 이 중 ${\alpha}$-glucosidase 저해활성이 가장 높은 오디와인으로부터 효소저해제를 분리 및 동정하기 위해 오디와인을 먼저 감압농축하여 에틸아세테이트로 분획한 다음 Diaion HP-20, ODS-A 및 Sephadex LH-20 column chromatography를 순차적으로 실시하여 4가지 분획을 얻었으며, 이 중 ${\alpha}$-glucosidase 저해활성이 가장 강한 Fr. 4로부터 UV 및 NMR 분석을 통해 quercetin를 동정하였다. 마지막으로 뽕나무 품종 및 발효에 따른 오디와인의 ${\alpha}$-glucosidase 저해제의 함량을 측정한 결과, 대성뽕 오디와인의 quercetin 함량(16.83 ppm)이 가장 높았으나, 익수뽕 및 청일뽕 오디와인의 quercetin 함량은 각각 14.85 및 8.92 ppm으로 낮았다. 그리고 발효 및 숙성에 따라 오디와인의 quercetin 함량은 크게 감소하였다. 이상의 연구 결과로부터 4가지 베리류 과실와인 중 오디와인이 가장 높은 ${\alpha}$-glucosidase 저해활성을 나타내었으며, 주된 저해물질은 quercetin임을 기기분석을 통해 확인할 수 있었기에 향후 오디와인은 당뇨예방용 기능성와인으로서 크게 각광을 받을 것으로 사료된다.

Keywords

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