中华鳖裙边胶原蛋白的提取、鉴定及其理化性质
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摘要
为增加中华鳖产品附加值,开发新型胶原蛋白资源,该研究以中华鳖裙边为原料,采用不同提取方式,分别提取酸溶性胶原蛋白(acid-soluble collagen,ASC)和胃蛋白酶可溶性胶原蛋白(pepsin-soluble collagen,PSC),并对2种胶原蛋白结构和理化性质进行分析。利用扫描电子显微镜(scanning electron microscope,SEM)、圆二色光谱(circular dichroism specturm,CD)对胶原蛋白的结构进行分析,采用SDS-PAGE凝胶电泳、肽指纹质谱(matrix-assisted laser desorption ionization-time of flight,MALDI-TOF)和氨基酸分析仪对ASC和PSC进行蛋白质和氨基酸比较分析,利用差示量热扫描(differential calorimetry scanning,DSC)对胶原蛋白进行热稳定性分析。结果表明,ASC和PSC的得率分别为26. 79%和37. 50%; PSC与ASC相比存在更完整的三级螺旋结构; ASC和PSC均为I型胶原蛋白,且具有较高的亚氨基酸和精氨酸含量; ASC和PSC的热变性温度分别为38. 39和38. 11℃,高于常见的猪皮和鱼皮胶原蛋白,具有较好的热稳定性。该研究为深入研究和开发中华鳖的裙边资源提供了理论依据。
Acid-soluble collagen(ASC) and pepsin-soluble collagen(PSC) were extracted from Chinese sturgeon calipash to add values for Chinese sturgeon products and develop new collagen resources. The structural properties of ASC and PSC were analyzed by scanning electron microscope(SEM) and circular two color spectrum(CD).The protein and amino acids of ASC and PSC were analyzed by SDS-PAGE gel electrophoresis,peptide mass spectrometry(MALDI-TOF) and amino acid analyzer. Differential scanning calorimetry(DSC) was performed to examine the thermal stabilities of ASC and PSC. The results showed that the yields of ASC and PSC were 26. 79% and37. 50%,respectively. Moreover,PSC had a more complete tertiary spiral structure than ASC did,and both ASC and PSC were type I collagen with high sub-amino acid and arginine contents. Furthermore,both ASC and PSC had better thermal stability as their thermal denaturation temperatures were 38. 39 and 38. 11 ℃,respectively,which were higher than that of common pig skin and fish skin collagen. This study provides a theoretical basis for in-depth research and development of calipash of Chinese sturgeon.
Acid-soluble collagen(ASC) and pepsin-soluble collagen(PSC) were extracted from Chinese sturgeon calipash to add values for Chinese sturgeon products and develop new collagen resources. The structural properties of ASC and PSC were analyzed by scanning electron microscope(SEM) and circular two color spectrum(CD).The protein and amino acids of ASC and PSC were analyzed by SDS-PAGE gel electrophoresis,peptide mass spectrometry(MALDI-TOF) and amino acid analyzer. Differential scanning calorimetry(DSC) was performed to examine the thermal stabilities of ASC and PSC. The results showed that the yields of ASC and PSC were 26. 79% and37. 50%,respectively. Moreover,PSC had a more complete tertiary spiral structure than ASC did,and both ASC and PSC were type I collagen with high sub-amino acid and arginine contents. Furthermore,both ASC and PSC had better thermal stability as their thermal denaturation temperatures were 38. 39 and 38. 11 ℃,respectively,which were higher than that of common pig skin and fish skin collagen. This study provides a theoretical basis for in-depth research and development of calipash of Chinese sturgeon.
引文
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[2]DU M,MCCORMICK R J.Applied Muscle Biology And Meat Science[M].CRC Press,2009.
[3]韩霜,马良,王雪蒙,等.兔皮胶原蛋白的加工特性[J].食品科学,2017,38(5):21-25.
[4]张丹.中华鳖营养、风味及功能特性研究进展[J].食品工业科技,2013,34(24):392-395.
[5]陆剑锋,万全,殷章敏,等.中华鳖裙边胶原蛋白的提取及其特征[J].水产学报,2010,34(6):801-808.
[6]LIU H Y,LI D,GUO S D.Studies on collagen from the skin of channel catfish(Ictaluru spunctaus)[J].Food Chemistry,2007,101(2):621-625.
[7]汪海波,梁艳萍,汪海婴,等.草鱼鱼鳞胶原蛋白的提取及其部分生物学性能[J].水产学报,2012,36(4):553-561.
[8]WANG J,PEI X,LIU H,et al.Extraction and characterization of acid-soluble and pepsin-soluble collagen from skin of loach(Misgurnus anguillicaudatus)[J].International Journal of Biological Macromolecules,2017,106.
[9]LAEMMLI U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227(5259),680-685.
[10]HEU M S,KIM J S,SHAHIDI F.Components and nutritional quality of shrimp processing by-products[J].Food Chemistry,2003,82(2):235-242.
[11]WANG L,LIANG Q,CHEN T,et al.Characterization of collagen from the skin of Amur sturgeon(Acipenser schrenckii)[J].Food Hydrocolloids,2014,38:104-109.
[12]PORTIER R J,MOODY M W,OGAWA M.Biochemical properties of black drum and sheepshead seabream skin collagen[J].J.agric.food Chem,2003,51(27):8 088-8 092.
[13]NALINANON S,BENJAKUL S,KISHIMURA H,et al.Type I collagen from the skin of ornate threadfin bream(Nemipteru shexodon):Characteristics and effect of pepsin hydrolysis[J].Food Chemistry,2010,125(2):500-507.
[14]CHEN J,LI L,YI R,et al.Extraction and characterization of acid-soluble collagen from scales and skin of tilapia(Oreochromis niloticus)[J].LWT-Food Science and Technology,2016,66:453-459.
[15]STEPLEWSKI A,MAJSTEREK I,MCADAMS E,et al.Thermostability gradient in the collagen triple helix reveals its multi-domain structure[J].Journal of Molecular Biology,2004,338(5):989-998.
[16]史刘辉.鳕鱼皮胶原及其低聚肽的制备及特性研究[D].无锡:江南大学,2012.
[17]USHA R,RAMASAMI T.Structure and conformation of intramolecularly cross-linked collagen[J].Colloids&Surfaces B Biointerfaces,2005,41(1):21-24.
[18]邹明晖,周建良,陈义初,等.枝化状聚乙二醇对牛跟腱I型胶原结构与构象稳定性的影响[J].中华实验外科杂志,2011,28(6):973-975.
[19]杨玲,赵燕,鲁亮,等.鲟鱼鱼皮胶原蛋白的提取及其理化性能分析[J].食品科学,2013,34(23):41-46.
[20]邓明霞,汪海波,杨玲,等.氨基酸组成及溶剂环境对淡水鱼胶原蛋白热稳定性能的影响[J].现代食品科技,2015(12):111-120.
[21]林琳.鱼皮胶原蛋白的制备及胶原蛋白多肽活性的研究[D].青岛:中国海洋大学,2006.
[22]李永明,徐子伟.差示扫描量热法(DSC)评定豆粕产品质量初探[J].中国粮油学报,2003,18(5):82-84.
[23]WANG L,YANG B,WANG R,et al.Extraction of pepsin-soluble collagen from grass carp(Ctenopharyngodon idella)skin using an artificial neural network[J].Food Chemistry,2008,111(3):683-686.
[24]JONGJAREONRAK A,BENJAKUL S,VISESSANGUANW,et al.Isolation and characterisation of acid and pepsin-solubilised collagens from the skin of Brownstripe red snapper(Lutjanus vitta)[J].Food Chemistry,2006,93(3):475-484.
[25]NAGAI T,YAMASHITA E,TANIGUCHI K,et al.Isolation and characterization of collagen from the outer skin waste material of cuttlefish(Sepia lycidas)[J].Food Chemistry,2001,72(4):425-429.
[26]王雪蒙,于玮,马良,等.兔皮胶原蛋白的提取及其结构鉴定[J].食品与发酵工业,2016,42(4):209-213.