3种不同方式提取鳕鱼鱼鳔胶原蛋白与胶原蛋白肽的基本特性研究
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摘要
目的研究3种不同提取方式所制备的大西洋鳕鱼(Gadus macrocephalus)鱼鳔胶原蛋白与胶原蛋白肽的基本特性。方法采用低温酸溶提取酸溶性胶原蛋白(acid-soluble collagen,ASC),热水抽提明胶(gelatin,GEL),复合蛋白酶酶解制备鳕鱼鱼鳔肽(swim bladder peptides,SWP)对鳕鱼鱼鳔中胶原蛋白的特性进行研究。对提取的胶原蛋白与胶原蛋白肽进行感官评定、SDS-PAGE电泳分析、紫外光谱分析、傅里叶红外光谱分析、氨基酸组成分析与扫描电镜分析。结果 ASC、GEL与SWP 3种胶原蛋白与胶原蛋白肽的提取率分别为42.15%、72.20%与89.11%,且羟脯氨酸的含量为8.73%、8.97%与7.94%;SDS-PAGE图谱显示ASC至少由2条α链与1条β链组成,GEL中存在少量的α链,SWP中α链与β链被降解。紫外光谱结果显示3种蛋白在230nm波长处均有最大吸收;扫描电镜结果显示ASC与GEL具有一定的交联结构,SWP中无交联结构存在。结论 3种不同提取方式制备的胶原蛋白与胶原蛋白肽具有不同的特性。
Objective To study the characterizations of collagen from Atlantic cod swim bladder prepared by three different extraction methods.Methods Acid soluble collagen(ASC)was extracted by acid soluble method at low temperature;Gelatin(GEL)was extracted by hot water;Swim bladder peptides(SWP)were extracted by enzymatic hydrolysis with protamex.The products were characterized by sensory evaluation,SDS-PAGE electrophoretic analysis,ultraviolet spectral analysis,Fourier infrared spectroscopy(FT-IR),amino acid composition analysis and Scanning electron microscopy(SEM).Results The extraction rates of three kinds of collagen were 42.15%,72.20%and 89.11%,respectively.The contents of hydroxyproline of them were 8.73%,8.97%and 7.94%.The atlas of SDS-PAGE showed that ASC was composed of at least twoαchains and oneβchain;the GEL contained a small number ofαchains;theαandβchains were degraded in SWP.The results of scanning electron microscopy showed that ASC and GEL had a certain cross-linking structure and there was no cross-linking structure in SWP.Conclusion The collagens prepared by three kinds of extraction methods had different characteristics.
Objective To study the characterizations of collagen from Atlantic cod swim bladder prepared by three different extraction methods.Methods Acid soluble collagen(ASC)was extracted by acid soluble method at low temperature;Gelatin(GEL)was extracted by hot water;Swim bladder peptides(SWP)were extracted by enzymatic hydrolysis with protamex.The products were characterized by sensory evaluation,SDS-PAGE electrophoretic analysis,ultraviolet spectral analysis,Fourier infrared spectroscopy(FT-IR),amino acid composition analysis and Scanning electron microscopy(SEM).Results The extraction rates of three kinds of collagen were 42.15%,72.20%and 89.11%,respectively.The contents of hydroxyproline of them were 8.73%,8.97%and 7.94%.The atlas of SDS-PAGE showed that ASC was composed of at least twoαchains and oneβchain;the GEL contained a small number ofαchains;theαandβchains were degraded in SWP.The results of scanning electron microscopy showed that ASC and GEL had a certain cross-linking structure and there was no cross-linking structure in SWP.Conclusion The collagens prepared by three kinds of extraction methods had different characteristics.
引文
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[4]Onouma K,Soottawat B,Thammarat K,et al.Characteristics of collagens from the swim bladders of yellowfin tuna(Thunnus albacares)[J].Food Chem,2014,155:264-270.
[5]王珊珊.鳕鱼骨胶原肽与活性钙的制备及其抗骨质疏松活性研究[D].青岛:中国海洋大学,2013.
[6]王杉杉.牦牛皮胶原蛋白的提取及其活性肽的制备[D].绵阳:西南科技大学,2018.
[7]孙晓飞,蒋丹,万超,等.建立一种快速鉴定大西洋鳕鱼及其制品的SYBR Green荧光PCR方法[J].东北师大学报(自然科学版),2017,49:127-130.
[8]张宝,陈运中,徐颖,等.草鱼鱼鳔胶原性质的研究[J].食品科学,2010,31(1):58-62.
[9]宁正祥.食品成分分析手册[M].北京:中国轻工业出版社,1998:163-165.
[10]Liu D S,Liang L,Regenstein J M,et al.Extraction and characterisation of pepsin-solubilised collagen from fins,scales,skins,bones and swim bladders of bighead carp(Hypophthalmichthys nobilis)[J].Food Chem,2014,133:1441-1448.
[11]Ye M,Guo G Y,Lu Y,et al.Purification,structure and anti-radiation activity of melanin from Lachnum YM404[J].Int J Biol Macromol,2018,63:170-176.
[12]Dou J,Meng Y H,Liu L,et al.Purification,characterization and antioxidant activities of polysaccharides from thinned-young apple[J].Int J Biol Macromol,2015,72:31-40.
[13]Farvin K H S,Andersen L L,Otte J,et al.Antioxidant activity of cod(Gadus morhua)protein hydrolysates:Fractionation and characterization of peptide fractions[J].Food Chem,2016,204:409-419.
[14]王杉杉,罗学刚,苏峰丙,等.牦牛皮胶原蛋白的提取及性能的分析[J].精细化工,2018,35(5):830-837.
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[16]Li H,Liu B,Gao L Z,et al.Studies on bullfrog skin collagen[J].Food Chem,2004,84(1):65-69.
[17]Li Z R,Wang B,Chi C F,et al.Isolation and characterization of acid soluble collagens and pepsin soluble collagens from the skin and bone of Spanish mackerel(Scomberomorous niphonius)[J].Food Hydrocolloids,2013,31(1):103-113.
[18]Veeruraj A,Arumugam M,Balasubramanian T.Isolation and characterization of thermostable collagen from the marine eel-fish(Evenchelys macrura)[J].Process Biochem,2013,48(10):1592-1602.
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