Lactobacillus plantarum M23 균주를 이용한 Tyrosinase 저해 활성 발효유 생산의 최적화

임상동,김기성,Lim, Sang-Dong,Kim, Kee-Sung 한국축산식품학회 2012 한국축산식품학회지 Vol.32 No.5

본 연구는 tyrosinase 활성 저해효과가 있는 젖산균인 L plantarum M23 균주를 이용하여 멜라닌 형성을 억제하기 위한 최적 발효유 제조조건을 설정하는데 그 목적이 있다. 3수준 4인자 중심합성 계획법을 통해 실험설계하여 반응표면분석법을 사용하였다. 독립변수로는 yeast extract 농도(%, $X_1$), 포도첨가량(%, $X_2$), 배양온도($^{\circ}C$, $X_3$), 배양시간(h, $X_4$)을 설정하였고, 종속변수로는 pH(pH, $Y_1$), 종합적 기호도(score, $Y_2$)와 tyrosinase inhibitory activity(%, $Y_3$)를 설정하였으며, tyrosinase inhibitory activity가 가장 높고 pH는 4.4를 동시에 만족하는 최적화를 실시한 결과 yeast extract 농도는 0.13%, 포도 첨가량은 2.95%, 배양 온도는 $37.1^{\circ}C$, 배양 시간은 14.8 h일 때 예상되는 pH는 4.42, 기호도는 7.06, tyrosinase inhibitory activity은 86.65%이었으며, 실제 실험 결과 pH는 4.35, 기호도는 6.86, tyrosinase inhibitory activity은 84.05%로서 큰 차이를 나타내지 않았다. 이러한 결과를 토대로 melanin 생합성과정의 key enzyme인 tyrosinase을 활성억제하는 L. plantarum M23을 이용한 발효유는 피부의 미백 작용과 노화 억제 작용에 기여할 것으로 사료되었다. The melanin pigment in human skin is a major defense mechanism against ultraviolet light to the skin, but darken skin color. Tyrosinase is mainly responsible for melanin biosynthesis (melanogenesis) in animals and enzymatic browning (melanosis) in plants. The purpose of this study was to optimize the fermented milk process for the melanin formation inhibition by using Lactobacillus plantarum M23 with tyrosinase inhibitory activity. We used 4-factor-3-level central composite design combining with response surface methodology. Yeast extract concentration (%, $X_1$), addition of grape (%, $X_2$), incubation temperature ($^{\circ}C$, $X_3$) and incubation time (h, $X_4$) was used as an independent factor, on the other hand, pH (pH, $Y_1$), overall palatability (score, $Y_2$) and tyrosinase inhibitory activity (%, $Y_3$) was used as a dependant factor. Based on the optimization for the highest tyrosinase inhibitory activity with pH 4.4, the expected data of pH, palatability and tyrosinase inhibitory activity with 14.8 h incubation at $37.1^{\circ}C$ by the addition of 0.127% of yeast extract, 2.95% of grape was 4.42, 7.06 and 86.65%, but the real data was 4.35, 6.86 and 84.05%, respectively. Based on the previous results, fermented milk using Lactobacillus plantarum M23 with the tyrosinase inhibitory activity could contribute for the whitening and antiaging of human skin.

Tyrosinase Inhibitory Activity of Rosmarinic Acid on Human Skin Melanoma Cells Treated with Hydrogen Peroxide

Jeong-Ran Kang,Sun-Mi Yoo 한국실험동물학회 2009 Laboratory Animal Research Vol.25 No.2

The purpose of this study was to evaluate the tyrosinase inhibitory activity of rosmarinic acid on the cultured human skin melanoma cells damaged by reactive oxygen species (ROS). The cytotoxicity effect was analyzed by MTT assay for cell viability, tyrosinase inhibitory activity after treating the cells with or without hydrogen peroxide (H₂O₂) for 1-10 hours. Light microscopic study was also carried out for morphological changes of the cells and tyrosinase activities. H₂O₂ decreased the cell viability in the cultured human melanoma cells in a dose- and time-dependent manner after cells had been treated with 20-80 ?M of H₂O₂ for 7 hours. IC?? value of H₂O₂ by the MTT assay was 80 ?M. Rosmarinic acid increases cell viability, inhibitory activity of tyrosinase. In light microscopic study, the number of human skin melanoma cells treated with H₂O₂ was decreased compared with control, and H2O2 activated tyrosinase activity, which was observed through darkened cell color (mid-activity, +++). However, colors of cells treated with Rosmarinic acid changed from white to brown in cell staining due to tyrosinase activity (moderate-activity, ++). The treatment of the cells with H₂O₂ shows highly toxic effects on cultured human skin melanoma cells. Rosmarinic acid increases cell viability and tyrosinase inhibitory activity in the cells treated with H₂O₂. It is considered that Rosmarinic acid shows tyrosinase inhibitory effect on ROS such as H₂O₂.

오미자 발효액의 항산화 및 항균 활성과 미용효과

조은경,조혜은,최영주 한국응용생명화학회 2010 Journal of Applied Biological Chemistry (J. Appl. Vol.53 No.4

This study was aimed to investigate the contents of chemical components and the biological activity of fermented Omija (Schizandra chinensis Baillon.) beverage (FOB), which have not been well known up to now. General compositions and the contents of mineral and vitamin of FOB were lower than Omija before fermentation (OBF). In addition, most sucrose was digested into glucose and fructose in FOB. This suggests that the beneficial contents may be made from general compositions by fermentation, though more evidences are needed. Antioxidant activity of FOB was measured by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging activity and superoxide dismutase-like activity (SODA). DPPH radical scavenging activity and SODA were increased in a dose-dependent manner, and were about 79.7% and 60.8% at four times diluted FOB (25% of FOB), respectively. The FOB also showed strong antibacterial activity on Escherichia coli. Beauty effect of FOB was demonstrated by the analysis of tyrosinase and elastase inhibitory activities, and they were remarkably increased in a dose-dependent manner. Tyrosinase inhibitory activity was about 82% at two times diluted FOB (50% of FOB) and elastase inhibitory activity was 78.2% at five times diluted FOB (20% of FOB). These results suggest that FOB has a strong antioxidant activity, and tyrosinase and elastase inhibitory effects. 본 연구는 전통발효공정에 따라 오미자를 발효시켜 제조한 오미자 발효액의 여러 가지 생리활성에 대하여 조사하였다. 우선, 오미자 발효액의 일반성분, 무기질 및 비타민 분석을 시행한 결과에서 발효전의 오미자보다 탄수화물과 비타민 C 함량이 높게 나타났다. 또한, 주요 맛 성분인 유리당 함량을 측정한 결과, 오미자 발효액에서 glucose와 fructose 함량이 발효 전의 오미자 보다 높게 나타났다. 이에 반해, 발효 전 오미자에서 검출되는 sucrose가 오미자 발효액에서는 나타나지 않았다. 오미자 발효액의 항산화 효과를 분석하기 위해 DPPH radical 소거능을 측정한 결과, 4배 희석한 오미자 발효액(25%)에서 79.7%의 DPPH radical 소거능을 나타내었는데, 0.01%의 BHA 와 유사한 항산화력이 측정되었다. 또한 오미자 발효액의 SOD 활성은 발효원액에서 94.7%의 활성을 보였는데 대조구인 Vit C의 경우 농도 0.1%일 때 98.1%로 보고되어 있어 오미자 발효원액(100%)과 유사한 항산화능을 나타내고 있다. 이것으로 볼 때 오미자 발효액의 항산화 활성이 아주 높은 것으로 항산 화력에 관한 높은 이용가치를 의미한다. 또한, 오미자 발효액의 항균활성을 관찰한 결과 오미자 발효원액(100%)에서 E. coli에 대한 항균효과가 나타났다. Tyrosinase의 저해효과는 시료의 농도가 증가함에 따라 유의적으로 증가하는 경향을 나타내었는데 50% 농도의 오미자 발효액에서 82%의 높은 tyrosinase 억제효과를 나타내었다. 오미자 발효액의 피부 주름 예방효과를 측정하기 위하여 elastase 저해효과 실험을 실시한 결과 전반적으로 농도가 높아짐에 따라 비례적으로 저해활성이 증가하였다. 이상의 결과는 오미자 발효액의 우수한 생리활성을 증명하고 있고, 항산화능, 항균력, 피부미용 효과가 높은 것으로 나타나 기능성 음료의 소재로서 그 활용도가 높을 것으로 판단된다.

초음파 처리 키토산과 메일라드 반응액 혼합물에 의한 tyrosinase 억제 활성 및 항균력 분석

김경자(Kim, Kyoung-Ja),양용준(Yang, Yong-Joon) 한국산학기술학회 2010 한국산학기술학회논문지 Vol.11 No.7

본 연구의 목적은 초음파 처리 키토산과 메일라드 반응액 혼합물에 의한 tyrosinase 억제 활성과 항균력을 조사하고자 하였다. 감자로부터 tyrosinase를 분리하여 SDS-PAGE후 일반 단백질 염색과 tyrosinase 활성 염색을 행하 여 tyrosinase를 확인하였다. 여러 가지 당 (glucose, fructose, galactose, xylose, arabinose or ribose)과 cystein과의 반응 으로 생성된 메일라드 반응 산물의 tyrosinase 억제 활성을 비교한 결과, glucose과 cystein 반응으로 생성된 메일라드 반응 산물의 tyrosinase 억제 활성이 가장 높은 것으로 나타났다. 0.5% 아세트산에 녹인 1% 키토산을 시간별로 초음 파 처리한 후 항균력을 조사한 결과 1 시간 이상 초음파 처리한 경우 E. coli 균주 와 S. aureus 균주에서 억제 활성 을 보였다. 갈변 억제 활성과 항균력을 가진 혼합물 제조를 위하여 초음파 처리 키토산과 메일라드 반응액을 혼합비 를 달리하여 조사한 결과 부피 비로 1 : 1 로 혼합한 경우가 tyrosinase 억제 활성과 항균력이 가장 효과가 높은 것으 로 조사되었다. The aim of this study was to investigate the tyrosinase inhibitory activity and antimicrobial activity by mixtures of ultrasonicated chitosan and Maillard reaction products. Analysis of tyrosinase was purified from potato and confirmed by active staining after SDS-PAGE. Maillard reaction products (MRPs) were formed from various sugars (glucose, fructose, galactose, xylose, arabinose or ribose) and cystein. MRPs inhibited the tyrosinase purified from potato. The highest tyrosinase inhibitory activity was shown by MRP from glucose and cystein. Ultrasonicated chitosan (over 1 hr) showed antimicrobial activity at the concentration of 1% against E. coli and S. aureus. For the development of antibrowning agent with antimicrobial activity, tyrosinase inhibitory and antimicrobial activity by the mixtures of ultrasonicated chitosan and MRP were tested. 1:1 mixture of ultrasonicated chitosan and MRP from glucose and cystein was the best antibrowning agent having antimicrobial activity.

추출용매 및 발아시기에 따른 무순 추출물의 생리활성 비교

한진희(Jin-Hee Han),문혜경(Hye-Kyung Moon),정신교(Shin-Kyo Chung),강우원(Woo-Won Kang) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.4

본 연구에서는 무순을 추출용매 및 발아시기에 따라 추출하여 아질산염 소거능, SOD 유사 활성, tyrosinase 저해 활성, xanthine oxidase 저해 활성 그리고 ACE 저해 활성을 측정하였다. 아질산염 소거능 측정에서는 pH 1.2의 조건에서 각 추출물들이 81.44~89.71%로 가장 높은 소거능을 나타내었고 pH 1.2와 pH 4.0의 조건에서 발아 4일째와 8일째의 무순 추출물이 발아 12일째의 무순 추출물보다 높은 소거능 효과를 나타내었다. 또 추출용매에 따라 소거능의 차이는 보였으나 물로 추출한 추출물이 상대적으로 다른 용매 추출물에 비해 소거능 효과가 좋은 것으로 나타났다. pH 6.0 조건에서는 에탄올 추출물이 16.12%의 활성을 보여 발아 12일째 무순의 에탄올 추출물과 메탄올 추출물의 소거능 효과와 유사한 결과를 보였다. 무순 추출물의 SOD 유사 활성은 4.57~27.05%의 범위를 보였고 비교물질인 L-ascorbic acid의 활성이 52.15%로 무순 추출물의 활성보다 높은 것으로 나타냈다. 추출물 중에서는 발아 8일째의 아세톤 추출물과 메탄올 추출물의 활성이 높은 것으로 나타났고 발아 12일째의 무순 추출물의 SOD 유사 활성은 4.57~15.59%로 현저히 감소하는 것으로 나타났다. Tyrosinase 저해 활성은 발아 8일째와 12일째의 무순 추출물의 활성이 좋았으며, 추출용매는 메탄올 추출물이 62.65~84.89% 측정되어 가장 좋은 저해 활성을 보였다. Xanthine oxidase 저해 활성은 발아 4일째의 무순 추출물이 21.26~29.52%로 아세톤 추출물이 가장 높은 저해능을 나타내었고 발아초기에 비해 발아가 진행됨에 따라 저해능은 감소하는 경향을 나타내었다. 발아시기와 추출용매에 따른 ACE 저해능은 12.48∼51.78%의 저해 활성을 보였으며, 발아 8일째의 에탄올 추출물이 51.78%로 각 추출물 중 가장 높은 저해 활성을 보였다. 이와 같은 생리활성 결과를 기초로 하여 천연 기능성 식품의 소재 발굴 및 식품 개발에 있어 연구 활용의 가능성이 있을 것으로 기대되며 일반적으로 섭취하여 큰 효능을 기대하기 어려운 무순을 유효성분만을 추출하여 소재화 하는 것이 더 효율적이라고 판단된다. 또한 현행 건강기능식품법에 따르면 물, 주정, 핵산 등의 추출용매를 사용하여 유효성분을 추출하는 것을 허용하고 있으므로 용매의 종류 및 추출방법에 대한 추가적인 연구를 통해 큰 기대 효과를 줄 수 있을 것이라 생각된다. This study extracted radish bud (Raphanus sativus L.) and investigated its nitrite scavenging activity, superoxide dismutases (SOD)-like activity, tyrosinase inhibition activity, xanthine oxidase inhibition activity, and angiotensin-converting-enzyme (ACE) inhibition activity according to extraction solvent and sprouting period. For nitrite scavenging activity, each extract recorded its highest level of 81.44∼89.71% at pH 1.2. Radish bud extracts on sprouting days 4 and 8 showed greater scavenging activities than those on sprouting day 12 at pH 1.2 and pH 4.0. There were differences in scavenging activity according to extraction solvent based on water extract exhibiting improved scavenging activity. Ethanol extract recorded scavenging activity of 16.12% at pH 6.0, which was similar to those of ethanol and methanol radish bud extracts on sprouting day 12. SOD-like activity of radish bud extracts was in the range of 4.57∼27.05%. For comparison purposes, SOD-like activity of L-ascorbic acid was 52.15%, which was higher than that of radish bud extracts. Acetone and methanol extracts showed high SOD-like activities on sprouting day 8. SOD-like activity of radish bud extracts on sprouting day 12 significantly decreased to 4.57∼15.59%. Radish bud extracts recorded good tyrosinase inhibitory activities on sprouting 8 and 12, whereas methanol extracts recorded the greatest tyrosinase inhibitory activity at 62.65∼84.89%. Radish bud extracts recorded xanthine oxidase inhibition activity of 21.26∼29.52% on sprouting day 4, and acetone extracts showed the highest level of xanthine oxidase inhibition activity. Xanthine oxidase inhibitory activity tended to decrease with sprouting period compared early on. ACE inhibitory activity was in the range of 12.48∼51.78% according to sprouting period and extraction solvent. Ethanol extracts on sprouting day 8 showed the highest ACE inhibitory activity of 51.78%. These results will hopefully contribute to research into the identification of materials and development of products for natural functional foods.

약용식물의 Tyrosinase, Hyaluronidase 저해효과 및 항산화 활성

차배천 한국생약학회 2011 생약학회지 Vol.42 No.1

This study was conducted to investigate tyrosinase inhibitory effect, hyaluronidase inhibitory effect and antioxidant activity by DPPH radical scavenging method on the MeOH extract of 50 species medicinal plant for screening of functional properties. As a result, Chaenomeles sinensis Koehne extract among 50 species medicinal plant turned out to be having tyrosinase,hyaluronidase inhibitory effect and antioxidant activity. The major component of tyrosinase and hyaluronidase inhibitory effect was isolated from EtOAc extract of Chaenomeles sinensis Koehne. And the component of antioxidant activity was isolated from n-BuOH extract of Chaenomeles sinensis Koehne. Their structure of compounds were identified as oleanolic acid and (-)-epicatechin by spectroscopic evidence, respectively.

우엉 뿌리 추출물의 항산화 및 Tyrosinase 저해 활성과 Phenolic Compound 분석

임도연,이경인 한국생약학회 2014 생약학회지 Vol.45 No.2

In this study, we investigated on antioxidative activities and tyrosinase inhibitory activities of methanol extract andits fractions from roots of Arctium lappa. The total phenolic compound and flavonoid content of the ethylacetate fraction wasfound to be 818.29 mg/g and 360.59 mg/g as the highest content. In the measurement of DPPH radical scavenging ability andtyrosinase inhibitory activity, the ethylacetate fraction was higher than the other fractions and the extract. In addition, comparativeanalysis of phenolic compounds by liquid chromatography-mass spectrometry (MS)/MS system under the multiplereactionmonitoring (MRM) with negative-ion electrospray ionization mode. The main phenolic compounds in the extract andfractions of roots from Arctium lappa were cynarin and chlorogenic acid. The main phenolic compound of the ethylacetate fractionwas cynarin. n-Butanol fraction had a significantly higher chlorogenic acid content than other samples. In conclusion,DPPH radical scavenging ability and tyrosinase inhibitory activity of the cynarin-riched ethylacetate fraction showed the highest activity.

Inhibition of Tyrosinase Activity by Polyphenols of Chestnut Leaf

Choi, Sang-Won,Kim, Hyo-Jin 대구효성가톨릭대학교 응용과학연구소 1998 응용과학연구논문집 Vol.6 No.2

밤잎으로부터 분리된 7가지 polyphenol 화합물의 tyrosinase 저해효과를 mushroom tyrosinase를 사용한 in vitro assay을 이용하여 측정하였다. 먼저 밤잎의 아세톤추출물을 에테르, 에틸아세테이트 및 부탄올 순서대로 용매 분획하여 에틸아세테이트 분획을 얻었다. 다음 이 분획을 Sep-pak C_18 cartridge, HPLC 및 TLC를 이용하여 7가지 페놀화합물, gallic acid(GA), methyl gallate(MG), (+)-catechin(C), (-)-epicatechin(EC), (-)-epicatechin gallate(ECg), (-)-epigallocatechin(EGC) 및 (-)-epigallocatechin gallate(EGCg)을 각각 분리하였다. GA 및 EC는 그들의 gallate 화합물, MG, ECg 및 EGCg 보다 효소 저해활성이 훨씬 낮았다. 그리고 비록 3개의 gallate 화합물에서 서로 유의적인 효소 저해활성의 차이는 보이지 않았지만, MG가 모든 화합물 중 가장 높은 저해활성을 나타내었다. 이러한 결과를 미루어 볼 때 밤잎추출물의 tyrosinase에 대한 강한 저해작용은 잎에 존재하는 3가지 gallate 화합물에 의해 기인된 것으로 생각된다. Tyrosinase inhibitory effect of seven different polyphenols isolated from chestnut(Castanea crenata) leaf was determined using commercially available mushroom tyrosinase in an in vitro assay. The acetone extract of chestnut leaf was partitioned with ethyl ether, ethyl acetate, and n-butanol, stepwisely. The EtOAc fraction was further fractionated and isolated by Sep-Pak c_18 cartridges, HPLC and TLC to give seven different polyphenols, such as gallic acid(GA), methyl gallate(MG), (+)-catechin(C), (-)-epicatechin(EC), (-)-epicatechin gallate(ECg), (-)-epigaalocatechin(EGC) and (-) -epigallocatechin gallate(EGCg). CA, C, and EC are much less tyrosinase inhibitory activity than their gallates, MG, ECg, and EGcg. Especially, methyl gallate showed the strongest inhibitory activity against mushroom tyrosinase, although there were no significant differences of inhibition among three gallate compounds. These results suggest that three gallates, such as methyl gallate, ECg, and EGCg may be mainly responsible for strong tyrosinase inhibitory action of chestnut leaf extract.

갈화 추출물 및 분획물의 피부관련 생리활성

이계영 ( Geai Young Lee ),임도연 ( Do Youn Im ) 한국미용학회 2012 한국미용학회지 Vol.18 No.4

In this study, we investigated antioxidative activity, antibacterial activity, nitric oxide (NO) production inhibitory activity and tyrosinase inhibitory activity of the methanol extract and its fractions from Puerariae Flos. The polyphenol and flavonoid content of the ethylacetate fraction of the extract was 142.47 mg/g and 42.86 mg/g as the highest values respectively. In the measurement of 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical scavenging ability and NO production inhibitory activity, the ethylacetate fraction of the extract was exhibited higher effect than other samples. Moreover, tyrosinase inhibitory activity of the ethylacetate fraction and butanol fraction of the extract showed higher activity than arbutin used as positive control. On the other hand, in antibacterial activity against Staphylococcus epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa by disc diffusion assay, the ethylacetate fraction of the extract showed the highest activity. In conclusion, the ethylacetate fraction of the methanol extract from Puerariae Flos could be applicable to functional materials for skin-related fields such as whitening, anti-inflammation against external stimuli and infection.

Antioxidant and Tyrosinase Inhibitory Activities of Different Parts of Oriental Cherry (Prunus serrulata var. spontanea)

Park, Ji-Won,Yuk, Hyun-Gyun,Lee, Seung-Cheol 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.2

Six different parts (branch, flesh, flower, fruit, leaf, and seed) of oriental cherry (Prunus serrulata var. spontanea) were extracted with ethanol or water, then total phenol content (TPC), antioxidant activity, and tyrosinase inhibitory activity of the extracts were evaluated. The ethanol extracts showed higher TPC and antioxidant activity than the water extracts regardless of parts. The ethanol extracts of leaf as well as branch possessed superior TPC and antioxidant activity. The highest tyrosinase inhibitory activity was found in ethanol extract of leaf. There was no dramatic difference of tyrosinase inhibitory activities according to parts of cherry. The results suggest that leaf and branch of oriental cherry could be a candidate for antioxidant and anti-whitening materials in food or cosmetic industries.