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광어껍질을 활용한 펩신가수분해물 제조공정 최적화와 피부건강 기능성
강유안(You-an Kang),진상근(Sang-Keun Jin),고종현(Jonghyun Ko),최영준(Yeung Joon Choi) 한국해양바이오학회 2022 한국해양바이오학회지 Vol.14 No.1
Low-molecular weight peptides derived from fish collagen exhibit several bioactivities, including antioxidant, antiwrinkle, antimicrobial, antidiabetic, and antihypertension effects. These peptides are also involved in triglyceride suppression and memory improvement. This study aimed to investigate the optimal processing condition for preparing low-molecular weight peptides from flounder skin, and the properties of the hydrolysate. The optimal processing conditions for peptic hydrolysis were as follows: a ratio of pepsin to dried skin powder of 2% (w/w), pH of 2.0, and a temperature of 50°C. Peptic hydrolysate contains several low-molecular weight peptides below 300 Da. Gly–Pro–Hyp(GPHyp) peptide, a process control index, was detected only in peptic hydrolysate on matrix-assisted laser desorption/ionization-time-of-flight(MALDI-TOF) spectrum. 2,2’-azinobis-(3-3-ethylbenzothiazolline-6- sulfonic acid(ABTS) radical scavenging activity of the peptic hydrolysate was comparable to that of 1 mM ascorbic acid, which was used as a positive control at pH 5.5, whereas collagenase inhibition was five times higher with the peptic hydrolysate than with 1 mM ascorbic acid at pH 7.5. However, the tyrosinase inhibition ability of the peptic hydrolysate was lower than that of arbutin, which was used as a positive control. The antibacterial effect of the peptic hydrolysate against Propionibacterium acne was not observed. These results suggest that the peptic hydrolysate derived from a flounder skin is a promising antiwrinkle agent that can be used in various food and cosmetic products to prevent wrinkles caused by ultraviolet radiations.
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광어껍질 펩신 가수분해물 유래 콜라게네이즈 저해 펩타이드의 서열분석과 분자도킹
김동환(Dong-Hwan Kim),강유안(You-An Kang),고종현(Jonghyun Ko),최영준(Yeung Joon Choi),진상근(Sang-Keun Jin) 한국식품영양과학회 2022 한국식품영양과학회지 Vol.51 No.4
본 연구는 광어껍질 pepsin 가수분해물의 collagenase 활성을 제해하는 펩타이드를 확인하고 collagenase 활성 저해능을 평가할 목적으로 수행하였다. Collagenase 저해 펩타이드는 이온교환, 겔 여과 및 역상크로마토그래피를 사용하여 정제하였다. MALDI-TOF 질량분석 스펙트럼에서 7개의 펩타이드 Gln-Phe(293 Da), Val-Ile-Cys-Glu(462 Da), Arg-Gly-Glu(360 Da), Val-Asp-Leu(345 Da), Gly-Pro-Met(303 Da), Gly-Ser-Ala-Pro-Glu(459 Da) 및 Arg-Leu(287 Da)를 확인하였다. LC-Q-TOF 질량분광계를 사용한 서열분석에서 6개의 비교적 긴 아미노산 서열의 펩타이드를 확인하였다. 펩타이드의 아미노산 서열은 Ⅰ형 콜라겐 알파-1 사슬(UniProtKB-Q5NT96)에서 확인하여 광어껍질에서 유래함을 검증하였다. 펩타이드 Val-Ile-Cys-Glu는 in silico 분자토킹에서 가장 높은 83.9의 GOLD score와 83.25±14.03%의 collagenase 저해 활성능을 보였다. 광어껍질 가수분해 제조공정 관리를 위한 지표물질로 Gly-Pro-Hyp의 정량법을 검정하였다. Pepsin 가수분해물 3 lot의 Gly-Pro-Hyp 평균 함량은 156 μg/g이었다. 이 같은 결과는 광어껍질 유래 pepsin 가수분해물이 저분자 펩타이드에 기인한 높은 collagenase 활성 저해능을 보이며 항주름 기능을 위한 식품첨가제와 기능성 식품 조성물로 사용될 수 있음을 제시한다. Bioactive peptides were isolated from the peptic hydrolysate of flounder skin using chromatographic methods, and their amino acid sequence was identified. The collagenase inhibitory ability of the hydrolysate was evaluated. Seven peptides, including Gln-Phe, Val-Ile-Cys-Glu, Arg-Gly-Glu, Val-Asp-Leu, Gly-Pro-Met, Gly-Ser-Ala-Pro-Glu, and Arg-Leu, were identified using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. Six peptides in the range of 6 to 18 amino acid residues were identified using LC/quadrupole time-of-flight (Q-TOF) mass spectrometry. All the peptides were verified on the type I collagen α-1 chain obtained from the Bastard halibut of Unitprot (www.uniprot.org) (code number: Q5NT96). Analysis of the seven peptides from the MALDI-TOF mass spectrometry was carried out using in silico simulation tools, and four peptides were further assessed for their potential collagenase inhibitory activities. The amino acid sequence of Val-Ile-Cys-Glu showed the highest GOLD score of 83.9 in in silico molecular docking and inhibited the collagenase activity up to 83% compared to control. Gly-Pro-Hyp, a quality index for the product, was also verified by the high-performance liquid chromatography (HPLC) method and measured 156 μg/g in the hydrolysate. These results suggest that the peptic hydrolysate from flounder skin contains small peptides with high collagenase inhibitory activity. This hydrolysate may find application as a food material or as an ingredient in skin products with an anti-wrinkle function.
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