文献类型：期刊文献

中文题名：基于分子对接的三角帆蚌酪氨酸酶抑制肽筛选及活性评价

英文题名：Screening and Activity Evaluation of Tyrosinase Inhibitory Peptides from Hyriopsis cumingii Based on Molecular Docking

作者：林海生[1,2];康佳傲[1];李菲[1,2];谢绍河[3];曾堉斯[1];秦小明[1,2];曹文红[1,2];陈忠琴[1,2];郑惠娜[1,2]

机构：[1]广东海洋大学食品科技学院,广东湛江524088;[2]广东省水产品加工与安全重点实验室/广东普通高等学校水产品深加工重点实验室/国家贝类加工技术研发分中心(湛江)/南海生物资源开发与利用协同创新中心,广东湛江524088;[3]广东绍河珍珠有限公司,广东汕头515041

年份：2024

卷号：44

期号：5

起止页码：133

中文期刊名：广东海洋大学学报

外文期刊名：Journal of Guangdong Ocean University

收录：北大核心2023、CSTPCD、、CSCD_E2023_2024、北大核心、CSCD

基金：财政部和农业农村部国家现代农业产业技术体系项目(CARS-49);广东省现代农业产业技术体系创新团队项目(2021KJ146);广东省大学生创新创业训练计划项目(S202310566017);广东海洋大学贝类高值化加工利用创新团队(CXTD2021004)。

语种：中文

中文关键词：三角帆蚌;珍珠层粉;酪氨酸酶抑制;活性肽;分子对接

外文关键词：Hyriopsis cumingii;nacre powder;tyrosinase inhibition;active peptides;molecular docking

中文摘要：【目的】筛选和鉴定三角帆蚌(Hyriopsis cumingii)珍珠层蛋白酶解产物中的酪氨酸酶抑制肽,为美白肽的研究与开发提供依据。【方法】以三角帆蚌珍珠层粉为原料,基于其蛋白组成特性,采用物理改性和可控酶解技术制备珍珠层活性肽(Peptides from nacre powder,P-NP),通过测定其2,2′-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)和1,1-二苯基-2-苦基肼自由基(DPPH)的清除能力、酪氨酸酶抑制活性以及对紫外线B(UVB)辐照损伤人永生化角质形成细胞(HaCaT)的影响,探究其体外活性。采用液质联用技术(LC-MS/MS)和分子对接技术从P-NP中鉴定并筛选出三角帆蚌酪氨酸酶抑制肽,并进行固相合成和体外活性验证。【结果】P-NP的ABTS和DPPH自由基清除效果较佳,肽质量浓度为1.00、14.00 mg/mL时,两者清除率分别为41.58%和11.23%;P-NP的酪氨酸酶活性抑制能力良好,其半抑制浓度(IC_(50))值为7.51 mg/mL。P-NP质量浓度在83.22μg/mL时,显著提高UVB损伤HaCaT细胞的生长活力(P<0.05)。P-NP经LC-MS/MS鉴定后筛选出相对分子质量集中在500~1000的61条肽序列作为候选肽库,分子对接筛选出3条潜在的酪氨酸酶抑制肽,其主要通过氢键与酪氨酸酶活性中心及其以外的位点相结合,从而达到抑制其活性的作用。其中,Tyr-Pro-Asn-Pro-Tyr(YPNPY)对酪氨酸酶抑制作用最强,IC_(50)为(0.545±0.028)mmol/L,主要抑制类型为竞争型抑制。【结论】三角帆蚌珍珠层活性肽具有良好的清除自由基和酪氨酸酶抑制活性。

外文摘要：【Objective】To identify and screen the whitening factor tyrosinase-inhibiting peptides from enzymatic hydrolysate of freshwater pearl mussels Hyriopsis cumingii,and provide a basis for the research and development of whitening peptides.【Method】The pearl active peptide(Peptides from nacre powder,P-NP)was prepared from the nacre powder of H.cumingii(NP-Hc)by physical modification and controlled enzymatic technology,based on the protein composition of the nacre powder,and its activity in vitro was explored by measuring the scavenging capacity of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium(ABTS)and 1,1-diphenyl-2-picrylhydrz(DPPH),and tyrosinase inhibitory capacity,and the effect of P-NP on the ultraviolet B(UVB)irradiation damaging human immortalised keratinocytes(HaCaT).Liquid chromatography-mass spectrometry(LC-MS/MS)and molecular docking techniques were used to identify and screen H.cumingii tyrosinase inhibitory peptides from P-NP,which were solid-phase synthesized and then verified for in vitro activity.【Result】P-NP had a good scavenging effect on ABTS and DPPH free radicals(41.58%and 11.23%at the concentration of 1.00 mg/mL and 14.00 mg/mL,respectively),and showed good inhibition of tyrosinase activity with an IC_(50) value of 7.51 mg/mL.P-NP(with peptide concentration of 83.22μg/mL)significantly increased the growth viability of UVB damaged HaCaT(P<0.05).Sixty-one peptide sequences,identified from P-NP by LC-MS/MS and with a relative molecular mass of 500-1000,were selected to establish a candidate peptide library.And three potential tyrosinase inhibitory peptides screened by molecular docking mainly bind to the active center of tyrosinase through hydrogen bond,thus achieving the inhibition activity.Among them,the peptide Tyr-Pro-Asn-Pro-Tyr(YPNPY)had the strongest inhibitory effect on tyrosinase,with an IC_(50) of(0.545±0.028)mg/mL and a competitive inhibition type.【Conclusion】The active peptides derived from pearl layer of H.cumingii have good free radical scavenging and tyrosinase inhibiting activities.