鱼皮生物活性肽的提取及功能活性研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2022-1265

摘要/Abstract

摘要：

对水产加工过程中的鱼皮废弃物进行深加工处理，对其高附加值利用可以变废为宝，减少环境污染；鱼皮来源生物活性肽具有抗氧化、抗高血压、抗菌等功效。对鱼皮废弃物中生物活性肽的提取、分离鉴定方法及其功能活性进行深入探讨，可为食品、保健品、化妆品、医药品和化工产品等相关产品的开发提供理论基础。本文综述了酶法、化学法及发酵法提取鱼皮来源生物活性肽的优缺点，酶法相对于化学法应用广泛，获得的生物活性肽活性高，发酵法成本低，适用于批量生产。归纳总结了超滤、纳滤、凝胶过滤、离子交换和高效液相色谱及质谱联用等鱼皮生物活性肽的分离纯化和鉴定方法，多种分离、鉴定方法相结合来获取具有特定功能活性的鱼皮生物活性肽是首选，但获得高纯度及高活性的目标产物仍是当前亟待突破的难点。此外，分析了鱼皮生物活性肽在抗氧化性、血管紧张素转换酶（angiotensin converting enzyme, ACE）抑制活性、抗菌性及其他生物活性的研究现状，归纳了功能活性与肽的分子量大小、肽序列结构及位置的构效关系。最后展望了生物活性肽在功能活性研发方面的不足之处及进一步研究的方向，以期可为鱼类加工业打造高值化利用功能性产品提供参考。

关键词: 鱼皮, 生物活性肽, 提取, 分离鉴定, 功能活性, 抗氧化性, ACE抑制活性, 抗菌性

Abstract:

Deep processing of fish skin waste in aquatic products processing can turn waste into treasure in high-value-added way and reduce environmental pollution, and the bioactive peptides from fish skin possess antioxidant, antihypertensive and antibacterial effects, etc. The methods of extraction, separation and identification of bioactive peptides from fish skin waste and their functional activities were discussed in depth, which may provide theoretical basis for the development of food, health products, cosmetics, pharmaceuticals and chemical products. In this paper, the advantages and disadvantages of enzyme, chemical and fermentation methods for the extraction of bioactive peptides from fish skin are reviewed. Compared with chemical methods, enzyme method is widely used, by which the bioactive peptides have higher activity; the cost of fermentation method is low, and it is suitable for mass production. The separation, purification and identification methods of fish skin bioactive peptides, such as ultrafiltration, nanofiltration, gel filtration, ion exchange, high performance liquid chromatography and mass spectrometry are summarized. The combination of multiple separation and identification methods to obtain fish skin bioactive peptides with specific functional activities is the first choice, but to obtain high purity and high activity of the target products is still a difficult breakthrough. In addition, the functional activities of fish skin bioactive peptides, such as antioxidant properties, inhibitory activities of angiotensin converting enzyme（ACE）, antibacterial properties, and other bioactivities are analyzed, and the structure-activity relationship between functional activities and molecular weight, sequence structure and location of the peptides is summarized. Finally, the deficiencies of functional activity research, the development of bioactive peptides, and the direction of further research are prospected, aiming to provide reference for the fish processing industry in making high-value functional products.

Key words: fish skin, bioactive peptide, extraction, isolation and identification, functional activity, antioxidant activity, ACE inhibitory activity, antibacterial activity

游子娟, 陈汉林, 邓辅财. 鱼皮生物活性肽的提取及功能活性研究进展[J]. 生物技术通报, 2023, 39(7): 91-104.

YOU Zi-juan, CHEN Han-lin, DENG Fu-cai. Research Progress in the Extraction and Functional Activities of Bioactive Peptides from Fish Skin[J]. Biotechnology Bulletin, 2023, 39(7): 91-104.

图/表 5

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常规提取方 Conventional extraction method	优点 Advantages	缺点 Disadvantages	文献来源 Reference
酶法 Enzymatic method	水解条件温和、反应安全可控，产物结构稳定，不破坏氨基酸，功能活性高，利于人体吸收 The hydrolysis conditions are mild, reactions are safe and controllable, the structure of the product is stable and it does not destroy amino acids, functional activity is high, and the hydrolysates are conducive to human absorption	酶活性较弱，反应速度慢，水解不彻底，价格昂贵、生产成本高 The activity of enzyme is weak, reaction speed is slow, the hydrolysis is not complete, the price of enzyme is expensive, and has high production cost	[12-13]
化学法 Chemical method	操作简便、成本低 Easy control, and low cost	引入有毒物质，安全性低，部分氨基酸被破坏，水解产物成分复杂，应用受限 It introduces toxic substances, safety is low, partial amino acids are destroyed, the hydrolysate composition is complex, and the application scope is limited	[4,14]
发酵法 Fermentation	增加产物风味、去腥，操作简便，可批量生产，成本低 Increase product flavor, reduce fishy, easy to operate, and can be mass production at low cost	产率低，纯化困难，部分产酶菌有毒 Low yield, difficult to be purified, and some bacteria of producing enzyme is toxic	[15]

常规提取方 Conventional extraction method	优点 Advantages	缺点 Disadvantages	文献来源 Reference
酶法 Enzymatic method	水解条件温和、反应安全可控，产物结构稳定，不破坏氨基酸，功能活性高，利于人体吸收 The hydrolysis conditions are mild, reactions are safe and controllable, the structure of the product is stable and it does not destroy amino acids, functional activity is high, and the hydrolysates are conducive to human absorption	酶活性较弱，反应速度慢，水解不彻底，价格昂贵、生产成本高 The activity of enzyme is weak, reaction speed is slow, the hydrolysis is not complete, the price of enzyme is expensive, and has high production cost	[12-13]
化学法 Chemical method	操作简便、成本低 Easy control, and low cost	引入有毒物质，安全性低，部分氨基酸被破坏，水解产物成分复杂，应用受限 It introduces toxic substances, safety is low, partial amino acids are destroyed, the hydrolysate composition is complex, and the application scope is limited	[4,14]
发酵法 Fermentation	增加产物风味、去腥，操作简便，可批量生产，成本低 Increase product flavor, reduce fishy, easy to operate, and can be mass production at low cost	产率低，纯化困难，部分产酶菌有毒 Low yield, difficult to be purified, and some bacteria of producing enzyme is toxic	[15]

来源 Species	提取方法 Extraction method	分离、鉴定方法 Separation and identification methods	水解度Degree of hydrolysis, DH/%	产率 Yield/%	自由基清除率 Free radical scavenging rate			氨基酸序列 Amino acid sequences	文献来源Reference
来源 Species	提取方法 Extraction method	分离、鉴定方法 Separation and identification methods	水解度Degree of hydrolysis, DH/%	产率 Yield/%	1,1-二苯基-2-三硝基苯肼自由基1,1-diphenyl-2-picrylhydrazyl, DPPH·（IC₅₀）	羟自由基 Hydroxyl free radical, ·OH（IC₅₀）	2'-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐自由基 ABTS	氨基酸序列 Amino acid sequences	文献来源Reference
巴沙鱼皮 Pangasius bocourti skin	碱性蛋白酶水解 Alcalase hydrolysis	超滤膜过滤、SDS-PAGE Ultrafiltration membrane filtration, SDS-PAGE	48.06±1.97	—	—	—	（45.98±0.04）μmol TE/g	—	[41]
黄鳍金枪鱼皮 Yellowfin Tuna（Thunnus albacares）skin	碱性蛋白酶水解 Alcalase hydrolysis	超滤膜过滤、SDS-PAGE Ultrafiltration membrane filtration, SDS-PAGE	—	52.7	—	—	—	—	[52]
石斑鱼皮 Seabass（Lates calcarifer）skin	碱性蛋白酶水解 Alcalase hydrolysis	交联葡聚糖G-25和超高效液相色谱-电喷雾离子质量-质谱/质谱 Sephadex G-25 column, UPLC-ESI-MS/MS	—	59.7	—	—	（2.59 ±0.04）mmol TE/μmol	Val-Leu-Gly-Pro-Phe（532.313 0 Da）	[54]
					—	—	（81.41±0.34）mmol TE/μmol	Gly-Leu-Phe-Gly-Pro-Arg（646.367 1 Da）
					—	—	（10.47±0.35）mmol TE/μmol	Gly-Ala-Thr-Gly-Pro-Gln-Gly-Pro-Leu-Gly-Pro-Arg（1 107.590 5 Da）
					—	—	（0.5±0.02）mmol TE/μmol	Gln-Leu-Gly-Pro-Leu-Gly-Pro-Val（780.461 4 Da）
罗非鱼皮 Tilapia（Oreochromis niloticus）skin	蛋白酶E、中性蛋白酶 Protease E and multifect neutral	超滤、葡聚糖凝胶柱层析G-25、反向高效液相色谱、Q-TOF MS/ESI Ultrafiltration, Sephadex G-25 gel filtration column, RP-HPLC and Q-TOF MS/ESI	22.11 ± 0.19	—	—	4.61 μg/mL	—	Glu-Gly-Leu（317.33 Da）	[55]
					—	6.45 μg/mL	—	Tyr-Gly-Asp-Glu-Tyr（645.21 Da）
马面鱼皮 Navodon Septentrio nalis（Thamnaco nus septentrionallis）skin	酸性蛋白酶和Protease A ‘Amano’2G Acid Protease and Protease A ‘Amano’2G	超滤和凝胶柱层析法，超高效液相色谱-质谱Ultrafiltration, gel filtration chromatography, and UPLC-MS	—	—	1.80 mg/mL	—	—	Asn-Glu-Gly-Ala-Cys-Gly/ Gly-Glu-Gly-Ala-Cys-Asn	[56]

来源 Species	提取方法 Extraction method	分离、鉴定方法 Separation and identification methods	水解度Degree of hydrolysis, DH/%	产率 Yield/%	自由基清除率 Free radical scavenging rate			氨基酸序列 Amino acid sequences	文献来源Reference
来源 Species	提取方法 Extraction method	分离、鉴定方法 Separation and identification methods	水解度Degree of hydrolysis, DH/%	产率 Yield/%	1,1-二苯基-2-三硝基苯肼自由基1,1-diphenyl-2-picrylhydrazyl, DPPH·（IC₅₀）	羟自由基 Hydroxyl free radical, ·OH（IC₅₀）	2'-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐自由基 ABTS	氨基酸序列 Amino acid sequences	文献来源Reference
巴沙鱼皮 Pangasius bocourti skin	碱性蛋白酶水解 Alcalase hydrolysis	超滤膜过滤、SDS-PAGE Ultrafiltration membrane filtration, SDS-PAGE	48.06±1.97	—	—	—	（45.98±0.04）μmol TE/g	—	[41]
黄鳍金枪鱼皮 Yellowfin Tuna（Thunnus albacares）skin	碱性蛋白酶水解 Alcalase hydrolysis	超滤膜过滤、SDS-PAGE Ultrafiltration membrane filtration, SDS-PAGE	—	52.7	—	—	—	—	[52]
石斑鱼皮 Seabass（Lates calcarifer）skin	碱性蛋白酶水解 Alcalase hydrolysis	交联葡聚糖G-25和超高效液相色谱-电喷雾离子质量-质谱/质谱 Sephadex G-25 column, UPLC-ESI-MS/MS	—	59.7	—	—	（2.59 ±0.04）mmol TE/μmol	Val-Leu-Gly-Pro-Phe（532.313 0 Da）	[54]
					—	—	（81.41±0.34）mmol TE/μmol	Gly-Leu-Phe-Gly-Pro-Arg（646.367 1 Da）
					—	—	（10.47±0.35）mmol TE/μmol	Gly-Ala-Thr-Gly-Pro-Gln-Gly-Pro-Leu-Gly-Pro-Arg（1 107.590 5 Da）
					—	—	（0.5±0.02）mmol TE/μmol	Gln-Leu-Gly-Pro-Leu-Gly-Pro-Val（780.461 4 Da）
罗非鱼皮 Tilapia（Oreochromis niloticus）skin	蛋白酶E、中性蛋白酶 Protease E and multifect neutral	超滤、葡聚糖凝胶柱层析G-25、反向高效液相色谱、Q-TOF MS/ESI Ultrafiltration, Sephadex G-25 gel filtration column, RP-HPLC and Q-TOF MS/ESI	22.11 ± 0.19	—	—	4.61 μg/mL	—	Glu-Gly-Leu（317.33 Da）	[55]
					—	6.45 μg/mL	—	Tyr-Gly-Asp-Glu-Tyr（645.21 Da）
马面鱼皮 Navodon Septentrio nalis（Thamnaco nus septentrionallis）skin	酸性蛋白酶和Protease A ‘Amano’2G Acid Protease and Protease A ‘Amano’2G	超滤和凝胶柱层析法，超高效液相色谱-质谱Ultrafiltration, gel filtration chromatography, and UPLC-MS	—	—	1.80 mg/mL	—	—	Asn-Glu-Gly-Ala-Cys-Gly/ Gly-Glu-Gly-Ala-Cys-Asn	[56]

来源 Species	提取方法 Extraction method	分离、鉴定方法 Separation and identification methods	水解度 Degree of hydrolysis，DH/%	产率 Yield/%	半抑制浓度IC₅₀/（μmol·L^-1）	氨基酸序列Amino acid sequences	文献来源Reference
太平洋鳕鱼皮 Pacific cord fish（Gadus macrocephalus）skin	酶法 Enzyme method	超滤、阴离子交换层析、反向高效液相色谱、质谱Ultrafiltration, anion exchange chromatography, RP-HPLC and MS	—	—	6.9	GASSGMPG（662 Da）	[61]
					14.5	LAYA（436 Da）
阿拉斯加鳕鱼皮 Alaska pollack skins	酶法 Enzyme method	Sephadex G-25凝胶柱层析、反向高效液相色谱和高效液相色谱-四极杆飞行时间质谱 Sephadex G-25 gel filtration chromatography, RP-HPLC and HPLC-Q-TOF-MS			108.5	GPLGVP（538.31 Da）	[6]
鳐鱼皮 Skate （Rajiformes）skin	酶法Enzyme method	Sephadex G-25凝胶柱层析、反向高效液相色谱、Q-TOF MS/ESI Sephadex G-25 gel filtration chromatography, RP-HPLC and Q-TOF MS/ESI	—	—	95	PGPLGLTGP（975.38 Da）	[36]
					148	QLGFLGPR（874.45 Da）
鳐科鱼皮 Okamejei kenojei skin	碱性蛋白酶水解 Alcalase hydrolysis	超滤膜过滤、Superdex^TM Peptide 10/300 GL凝胶柱层析 Ultrafiltration, Superdex^TM Peptide 10/300 GL gel filtration chromatography	—	—	4.22	LGPLGHQ（720 Da）	[62]
					3.09	MVGSAPGVL（829 Da）