학술논문
Leuconostoc mesenteroides 유래 글리칸수크레이즈를 이용한 하이드로퀴논 글리코사이드의 합성과 특성 연구 / Synthesis and Characterization of Hydroquinone Glycosides Using Glycansucrases from Leuconostoc mesenteroides
Document Type
Dissertation/ Thesis
Source
Subject
Language
English
Abstract
하이드로퀴논은 티로신(Tyrosine)이 멜라닌으로 대사되는 과정에 참여하는 효소인 티로시나제의 활성을 억제하여 멜라닌 합성을 저해하고 생성된 색소를 퇴화(퇴색)시키며, 멜라닌 세포의 괴사를 유도하여 결과적으로 멜라닌색소의 피부 침착을 방지한다.하지만 독성을 가지고 있어, 피부염 등의 질환을 유발한다. 본 연구에서는 하이드로퀴논에 글리코사이드를 결합한 두가지의 신규 배당체를 효소적으로 합성하였으며, 이 새로운 배당체의 기능성을 확인하였다.하이드로퀴논배당체 (hydroquinond fructoside, hydroquinone glucoside; HF, HG)는 Leuconostoc mesenteroides 으로부터 유래한 dextransucrase와 levansucrase를 이용하여 설탕과 하이드로퀴논의 수용체 반응을 통하여 합성하였다. HF의 최적 생산조건은 반응 표면 방법(Response Sulface methodology) 을 이용하여 세 가지 요소(설탕농도, 하이드로퀴논 농도, 효소활성)의 상관 관계를 확인하였다. 설탕 농도는 115 mM, 하이드로퀴논 농도는 350 mM, 효소활성은 0.70 unit 일 때 가장 많은 배당체가 생산되었다. HG의 최적 생산 조건은 설탕 농도는 215 mM, 하이드로퀴논 농도는 450 mM, 효소활성은 0.55 unit 이었다. 합성반응 후 설탕 및 글루코오스 제거는 부탄올 첨가 후 상층액(부탄올 층)을 실리카 겔 컬럼 크로마토그래피를 이용하여 분리 후 HPLC로 정제 정도를 확인 하였다. 하이드로퀴논 배당체의 구조 분석은 NMR (Nuclear magnetic resonance)을 이용하여 ¹H, ^(13)C, HSQC, ¹H -¹H COSY, 그리고 HMBC로 하였고, MALDI-TOF MS를 확인 하여 HF는 4-hydroxyphenyl-ß-D-fructofuranoside 임을 확인하였고, HG는 4-hydroxyphenyl-α-D-glucopyranoside 임을 확인하였다. 여러 가지의 생리활성 및 기능성 실험을 통하여 HG와 HF 가 미백기능 및 항 산화 작용, 아질산 소거능, 지질과산화 억제효과가 뛰어 남을 알 수 있었다. 이로부터 HF와 HG 는 화장품 및 의약품 원료로 활용 될 수 있을 것으로 사료된다.
Hydroquinone (HQ) functions as a skin whitening agent, but it has the potential to cause dermatitis. We synthesized a novel HQ fructoside (HF) and hydroquinone glucoside (HG or α-arbutin) as a potential skin whitening agent by reacting levansucrase and dextransucrase, respectively, from Leuconostoc mesenteroides with HQ as an acceptor and sucrose as a donor. The products were purified using butanol partition and silica-gel column chromatography. The structures of the purified HF and HG were determined by nuclear magnetic resonance and the molecular ion of the product was observed both at m/z 295 (C12 H16 O7 Na)^(+). The HF was identified as 4-hydroxyphenyl-β-D-fructofuranoside and HG was identified as 4-hydroxyphenyl-α-D-glucopyranoside. The optimum condition for HF synthesis was determined using a response surface methodology and the final optimum condition was 350 mM HQ, 115 mM sucrose, and 0.70 U/ml levansucrase, and the final HF produced was 1.09 g/l. The optimum condition for HG synthesis was determined using a response surface methodology and the final optimum condition was 450 mM HQ, 215 mM sucrose, and 0.55 U/ml dextransucrase; the final HG produced was 544 mg/l. Both HF and HG showed anti-oxidation activities and inhibition against tyrosinase. The IC_(50) of HF for DPPH scavenging activity was 5.83 mM, showing higher anti-oxidant activity compared to β-arbutin (IC_(50) = 6.04 mM). The Ki value of HF (0.67-mM) against tyrosinase was smaller than that of β-arbutin (K_(i) = 2.8 mM). The inhibition of lipid peroxidation by HF was 108.12% that of HQ (100%) and much higher than that of β-arbutin (0.81% of HQ). In case of HG, the IC_(50) of diphenylpicryl-hydrazyl scavenging activity was 3.85 mM indicating a greater anti-oxidant activity compared to β-arbutin (IC_(50) = 6.04 mM). HG-mediated inhibition of lipid peroxidation was 3.51% that of HQ (100%) and much higher than that of β-arbutin (0.81% of HQ).
Hydroquinone (HQ) functions as a skin whitening agent, but it has the potential to cause dermatitis. We synthesized a novel HQ fructoside (HF) and hydroquinone glucoside (HG or α-arbutin) as a potential skin whitening agent by reacting levansucrase and dextransucrase, respectively, from Leuconostoc mesenteroides with HQ as an acceptor and sucrose as a donor. The products were purified using butanol partition and silica-gel column chromatography. The structures of the purified HF and HG were determined by nuclear magnetic resonance and the molecular ion of the product was observed both at m/z 295 (C12 H16 O7 Na)^(+). The HF was identified as 4-hydroxyphenyl-β-D-fructofuranoside and HG was identified as 4-hydroxyphenyl-α-D-glucopyranoside. The optimum condition for HF synthesis was determined using a response surface methodology and the final optimum condition was 350 mM HQ, 115 mM sucrose, and 0.70 U/ml levansucrase, and the final HF produced was 1.09 g/l. The optimum condition for HG synthesis was determined using a response surface methodology and the final optimum condition was 450 mM HQ, 215 mM sucrose, and 0.55 U/ml dextransucrase; the final HG produced was 544 mg/l. Both HF and HG showed anti-oxidation activities and inhibition against tyrosinase. The IC_(50) of HF for DPPH scavenging activity was 5.83 mM, showing higher anti-oxidant activity compared to β-arbutin (IC_(50) = 6.04 mM). The Ki value of HF (0.67-mM) against tyrosinase was smaller than that of β-arbutin (K_(i) = 2.8 mM). The inhibition of lipid peroxidation by HF was 108.12% that of HQ (100%) and much higher than that of β-arbutin (0.81% of HQ). In case of HG, the IC_(50) of diphenylpicryl-hydrazyl scavenging activity was 3.85 mM indicating a greater anti-oxidant activity compared to β-arbutin (IC_(50) = 6.04 mM). HG-mediated inhibition of lipid peroxidation was 3.51% that of HQ (100%) and much higher than that of β-arbutin (0.81% of HQ).