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罗非鱼皮胶原肽的酪氨酸酶抑制活性与铜离子结合活性研究

宋玉琼 胡晓 相欢 陈胜军 李来好 杨贤庆 吴燕燕 马海霞

宋玉琼, 胡晓, 相欢, 陈胜军, 李来好, 杨贤庆, 吴燕燕, 马海霞. 罗非鱼皮胶原肽的酪氨酸酶抑制活性与铜离子结合活性研究[J]. 南方水产科学. doi: 10.12131/20220097
引用本文: 宋玉琼, 胡晓, 相欢, 陈胜军, 李来好, 杨贤庆, 吴燕燕, 马海霞. 罗非鱼皮胶原肽的酪氨酸酶抑制活性与铜离子结合活性研究[J]. 南方水产科学. doi: 10.12131/20220097
SONG Yuqiong, HU Xiao, XIANG Huan, CHEN Shengjun, LI Laihao, YANG Xianqing, WU Yanyan, MA Haixia. Study on tyrosinase inhibitory activity and Cu2+ binding activity of tilapia skin peptides[J]. South China Fisheries Science. doi: 10.12131/20220097
Citation: SONG Yuqiong, HU Xiao, XIANG Huan, CHEN Shengjun, LI Laihao, YANG Xianqing, WU Yanyan, MA Haixia. Study on tyrosinase inhibitory activity and Cu2+ binding activity of tilapia skin peptides[J]. South China Fisheries Science. doi: 10.12131/20220097

罗非鱼皮胶原肽的酪氨酸酶抑制活性与铜离子结合活性研究

doi: 10.12131/20220097
基金项目: 广东省重点领域研发计划资助项目 (2020B1111030004);财政部和农业农村部:国家现代农业产业技术体系 (CARS-46);广东省基础与应用基础研究基金项目 (2019A1515011588);广东省科技创新战略专项资金计划项目 (2018S0044);扬帆计划引进创新创业团队专项 (2015YT02H109);中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助 (2021SD06)
详细信息
    作者简介:

    宋玉琼 (1997—),女,硕士研究生,研究方向为水产品加工与质量安全、食品生物技术。E-mail: SYQ1010730529@163.com

    通讯作者:

    胡 晓 (1981—),男,研究员,博士,研究方向为水产品加工与质量安全、食品生物技术。E-mail: hnhuxiao@163.com

  • 中图分类号: TS 254.1

Study on tyrosinase inhibitory activity and Cu2+ binding activity of tilapia skin peptides

  • 摘要: 为探明罗非鱼 (Oreochromis spp.) 鱼皮胶原蛋白肽能否有效抑制黑色素的生成,采用3种酪氨酸酶抑制肽 (Tilapia skin tyrosinase inhibitory peptides, TSTIP) 开展了TSTIP的酪氨酸酶 (Tyrosinase, TYR) 抑制活性与铜离子 (Cu2+) 结合活性的相关性研究。结果显示,罗非鱼皮经碱性蛋白酶酶解4 h后的产物具有最高的TYR抑制活性和Cu2+结合活性,且两者呈极显著正相关 (R=0.856)。荧光光谱分析显示,TSTIP与TYR或Cu2+结合后其内源荧光吸收强度的下降趋势一致,同时紫外吸收增强,且最大吸收波长发生红移。傅里叶变换红外光谱分析显示,TYR与Cu2+主要通过羰基及氨基与TSTIP结合。圆二色谱分析显示,两种结合物中β-转角及无规则卷曲含量相对减少,β-折叠含量相对增多,该现象在TSTIP-Cu2+结合物中更加明显。结果表明,TSTIP结合TYR产生的结构变化与结合Cu2+相似,TSTIP可通过螯合其活性中心Cu2+来抑制其活性。
  • 图  1  不同酶解物的TYR抑制率与Cu2+结合率和相关性分析

    注:a. 酶解物的单酚酶抑制活性;b. 酶解物的二酚酶抑制活性;c. 酶解物的Cu2+结合率;d. TSTIP二酚酶抑制活性与Cu2+结合率和相关性分析;不同小写字母表示同种蛋白酶在不同酶解时间所得酶解物的抑制率差异显著 (P<0.05);不同大写字母表示不同蛋白酶在同一酶解时间所得酶解物的抑制率差异显著 (P<0.05)。

    Figure  1.  Tyrosinase inhibition rate and Cu2+ binding rate of different hydrolysates and correlation analysis

    Note: a. Monophenolase inhibitory activity of hydrolysates; b. Diphenolase inhibitory activity of hydrolysates; c. Cu2+ binding rate of hydrolysates; d. Correlation analysis between tyrosinase inhibitory activity and Cu2+ binding rate. Different lowercase letters indicate that the same protease at different hydrolysis time had significant difference (P<0.05), while different uppercase letters indicate that different proteases at the same hydrolysis time had significant difference (P<0.05).

    图  2  AH分子量分布和质量分数

    注:a. TSTIP的分子量分布;b. AH不同组分的质量分数。

    Figure  2.  Molecular weight distribution and mass fraction of AH

    Note: a. Molecular weight distribution of TSTIP; b. Relative amino acid mass fraction of AH.

    图  3  TSTIP-TYR及TSTIP-Cu2+的荧光光谱

    注:a、b分别为TSTIP与不同浓度的TYR或Cu2+结合后的荧光强度变化;c、d分别为TSTIP与TYR或Cu2+结合后荧光强度随时间的变化。

    Figure  3.  Fluorescence intensity of TSTIP-TYR and TSTIP-Cu2+

    Note: The a and b represente the changes in fluorescence intensity of TSTIP combining with TYR or Cu2+ at different concentration, respectively; the c and d represente the changes in fluorescence intensity of TSTIP combining with TYR and Cu2+ at different time, respectively.

    图  4  TSTIP-TYR和TSTIP-Cu2+的紫外与红外光谱分析

    注:a. TSTIP-TYR及TSTIP-Cu2+的紫外吸收光谱; b. TSTIP-TYR及TSTIP-Cu2+的红外吸收光谱。

    Figure  4.  UV and FTIR spectrum of TSTIP-TYR and TSTIP-Cu2+

    Note: a. UV spectrums of TSTIP-TYR and TSTIP-Cu2+; b. FTIR spectrums of TSTIP-TYR and TSTIP-Cu2+.

    图  5  TSTIP-TYR及TSTIP-Cu2+的圆二色谱

    注:a. TSTIP-TYR和TSTIP-Cu2+的圆二色谱图; b. TSTIP-TYR和TSTIP-Cu2+的二级结构;不同小写字母表示同一样品在不同二级结构的质量比例有显著性差异 (P<0.05);不同大写字母表示不同样品在同种二级结构的质量比例有显著性差异 (P<0.05)。

    Figure  5.  CD analysis of TSTIP-TYR and TSTIP-Cu2+

    Note: a. CD spectrum of TSTIP-TYR and TSTIP-Cu2+; b. Proportions of secondary structures of TSTIP-TYR and TSTIP-Cu2+. Different lowercase letters indicate that the mass proportion of the same sample in different secondary structures had significant difference (P<0.05). Different uppercase letters indicate that the mass proportion of different samples in the same secondary structure had significant difference (P<0.05).

    表  1  不同蛋白酶的酶解条件

    Table  1.   Enzymatic conditions for different proteases

    蛋白酶
    Protease
    酶解条件 Hydrolysis condition
    酶活力
    Enzymatic activity/
    (U·mg−1)
    pH温度
    Temperature/ ℃
    加酶量
    Enzyme content/%
    酶解时间
    Hydrolysis time/h
    碱性蛋白酶 Alcalase 200 9.0 50 2 1、2、4、6、8 
    木瓜蛋白酶 Papain 1 500 6.5 50 2 1、2、4、6、8 
    胰蛋白酶 Trypsin 250 8.0 37 2 1、2、4、6、8 
    下载: 导出CSV

    表  2  罗非鱼皮和TSTIP中氨基酸成分和质量分数

    Table  2.   Amino acids composition and mass fraction of tilapia skin and TSTIP

    氨基酸
    Amino acids
    氨基酸质量分数
    Amino acid mass fraction/%
    罗非鱼皮
    Tilapia skin
    罗非鱼片酪氨酸酶抑制肽
    TSTIP
    天冬氨酸 Asp 6.20±0.23f 6.00±0.28f
    谷氨酸 Glu 10.78±0.14d 10.65±0.21d
    赖氨酸 Lys 3.69±0.23g 3.62±0.26g
    组氨酸 His 0.90±0.10jk 0.87±0.25kl
    精氨酸 Arg 9.45±0.18e 9.20±0.23e
    色氨酸 Trp 0.15±0.20l 1.80±0.22i
    酪氨酸 Tyr 0.65±0.23k 0.61±0.18l
    苯丙氨酸 Phe 2.21±0.25i 2.11±0.15i
    甘氨酸 Gly 26.28±0.21a 27.70±0.21a
    丙氨酸 Ala 11.22±0.17c 11.16±0.23c
    缬氨酸 Val 2.21±0.21i 2.19±0.26i
    蛋氨酸 Met 1.18±0.17k 1.47±0.34ij
    异亮氨酸 Ile 1.17±0.24j 1.10±0.40jk
    脯氨酸 Pro 14.32±0.15b 13.68±0.23b
    亮氨酸 Leu 2.95±0.09h 2.92±0.22h
    苏氨酸 Thr 2.95±0.04h 2.94±0.17h
    丝氨酸 Ser 3.84±0.14g 3.72±0.32g
    胱氨酸 Cys 0.01±0.12l 0.04±0.13m
    注:不同小写字母表示同种原料在不同氨基酸种类的相对质量分数差异显著 (P<0.05)。 Note: Different lowercase letters indicate that there are significant differences in the relative mass fraction of the same material in different amino acids (P<0.05).
    下载: 导出CSV
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出版历程
  • 收稿日期:  2022-04-07
  • 修回日期:  2022-06-06
  • 录用日期:  2022-07-19
  • 网络出版日期:  2022-07-27

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