利用光谱技术分析加热温度对肌红蛋白结构的影响
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摘要
以肌红蛋白纯溶液体系为研究对象,利用紫外-可见吸收光谱、荧光光谱、圆二色(CD)光谱为分析手段,研究了60～80℃的加热处理对肌红蛋白结构的影响。结果表明,80℃加热时,肌红蛋白的紫外-可见吸收光谱特征峰吸光值发生显著降低(p <0.05),荧光光谱最大发射峰的荧光强度显著增加(p <0.05),表面疏水性显著增高(p <0.05),二级结构中的α-螺旋含量显著降低(p <0.05)。而60、70℃加热处理对肌红蛋白二、三结构的影响较小。肌红蛋白对于70℃及以下的加热具有一定的耐受性,但随着加热温度由60℃升高至80℃,肌红蛋白的变性程度、血红素脱离程度、三级结构展开程度以及二级结构的无序化均呈增大趋势。表明70℃以上的加热会严重影响肌红蛋白的结构。
The myoglobin pure solution system was used as the research object,the effect of heat treatment at 60～80 ℃ on the structure of myoglobin were investigated by ultraviolet-visible absorption spectroscopy,fluorescence and circular dichroism (CD) spectra.The results showed that,the absorbance of the characteristic peaks of myoglobin decreased significantly (p <0.05),and the fluorescence intensity of the maximum emission peak of the fluorescence spectrum increased significantly,and the α-helix content in the secondary structure decreased significantly (p < 0.05) when heated at 80 ℃. However,60,70 ℃treatments had little effect on the secondary and tertiary structure of myoglobin.Myoglobin was resistant to heating up to 70 ℃.As the heating temperature raised from 60 ℃ to 80 ℃,the denaturate degree of myoglobin,the detachment degree of hemoglobin,the expansion degree of tertiary structure,and the disordering of secondary structure were all increasing,which indicated that heating temperature above 70 ℃ would affect the structure of myoglobin seriously.
The myoglobin pure solution system was used as the research object,the effect of heat treatment at 60～80 ℃ on the structure of myoglobin were investigated by ultraviolet-visible absorption spectroscopy,fluorescence and circular dichroism (CD) spectra.The results showed that,the absorbance of the characteristic peaks of myoglobin decreased significantly (p <0.05),and the fluorescence intensity of the maximum emission peak of the fluorescence spectrum increased significantly,and the α-helix content in the secondary structure decreased significantly (p < 0.05) when heated at 80 ℃. However,60,70 ℃treatments had little effect on the secondary and tertiary structure of myoglobin.Myoglobin was resistant to heating up to 70 ℃.As the heating temperature raised from 60 ℃ to 80 ℃,the denaturate degree of myoglobin,the detachment degree of hemoglobin,the expansion degree of tertiary structure,and the disordering of secondary structure were all increasing,which indicated that heating temperature above 70 ℃ would affect the structure of myoglobin seriously.
引文
[1]姚国佳,杨建平,李梦云,等.不同日龄猪背最长肌中肌红蛋白及其肉色变化规律研究[J].郑州牧业工程高等专科学校学报,2015,35(2):4-7.
[2]胡煌,吕飞,丁玉庭.肉制品的呈色机理和色泽评定研究进展[J].肉类研究,2016,30(12):48-53.
[3]赵怀璞,郑建国,游国强,等.肌红蛋白功能研究进展[J].山西医药杂志,2017,46(11):1299-1302.
[4]汤祥明,金邦荃.高铁肌红蛋白还原酶及其与肉色稳定性的研究进展[J].畜牧与兽医,2007,39(11):71-74.
[5] Joseph P,Suman S P,Li S,et al. Massspectrometric characterizationand thermostability of turkey myoglobin[J].LWTFood Science and Technology,2010,43(2):273-278.
[6]Berisha A,Yasushi E,Fujimoto K.The effect of heat-induced structural changes on the prooxidant activity of myoglobin[J].Italian Journal of Food Science,2003,15(1):85-94.
[7]王琳可.火候对卤煮鸡腿质构、色泽的影响研究[D].郑州:河南农业大学,2015.
[8]黄明,黄峰,张首玉,等.热处理对猪肉食用品质的影响[J].食品科学,2009,30(23):189-192.
[9]黄甜,严成,黄业传,等.高压结合热处理对猪肉色泽的影响[J].食品工业科技,2015,36(2):85-89.
[10]李升升.热处理对牦牛肉品质的影响及其相关性分析[J].食品与机械,2016(4):207-210.
[11]张淼,王道营,张牧焓,等.一磷酸腺苷与肌动球蛋白相互作用的荧光光谱研究[J].食品科学,2016,37(17):32-37.
[12]张玉姣,唐乾,曹洪玉,等.酸诱导拥挤条件下肌红蛋白及突变体去折叠过程[J].物理化学学报,2013,29(8):1785-1792.
[13]Schmal H,Niemeyer P,Zwingmann J,et al.Relationships of hydrophobicity and net charge to the solubility of milk and soy proteins[J].Journal of Food Science,1985,50(2):486-491.
[14]吴丹,徐桂英.光谱法研究蛋白质与表面活性剂的相互作用[J].物理化学学报,2006,22(2):254-260.
[15]王斌.中药小分子与牛血清白蛋白的相互作用[D].武汉:中南民族大学,2011:39-41.
[16]张莹莹,曹洪玉,唐乾,等.不同类型表面活性剂与高铁肌红蛋白相互作用[J].物理化学学报,2011,27(12):2907-2914.
[17]Ladokhin A S. Fluorescence spectroscopy in peptide and protein analysis[M]. New York:Encyclopedia of Analytical Chemistry,John Wiley&Sons,2006:5762-5779.
[18]吴名草,金邦荃,陈许明,等.紫外照射下高铁肌红蛋白变化的同步荧光光谱指认[J].激光与光电子学进展,2013,50(5)197-203.
[19]王玮,葛毅强,陈颖,等.超高压对高铁肌红蛋白还原酶活性及二级结构的影响[J].中国食品学报,2015,15(10):134-140.
[20]程兵兵.hPRS1及其突变体圆二色光谱研究[D].合肥:中国科学技术大学,2007.
[21]Taheri-Kafrani A,Asgari-Mobarakeh E,Bordbar A K,et al. Structure-function relationship ofβ-lactoglobulin in the presenceof dodecyltrimethyl ammonium bromide[J]. Colloids and Surfaces B Biointerfaces,2010,75(1):268-274.
[22]Moriyama Y,Takeda K. Critical temperature of secondary structural change of myoglobin in thermal denaturation up to 130degrees C and effect of sodium dodecyl sulfate on the change[J].Journal of Physical Chemistry B,2010,114(7):2430.
[23]曹莹莹.超高压结合热处理对肌球蛋白凝胶特性的影响研究[D].南京:南京农业大学,2012.
[24]温斯颖.不同种类动物肌肉蛋白质消化产物比较研究[D].南京:南京农业大学,2015.
[2]胡煌,吕飞,丁玉庭.肉制品的呈色机理和色泽评定研究进展[J].肉类研究,2016,30(12):48-53.
[3]赵怀璞,郑建国,游国强,等.肌红蛋白功能研究进展[J].山西医药杂志,2017,46(11):1299-1302.
[4]汤祥明,金邦荃.高铁肌红蛋白还原酶及其与肉色稳定性的研究进展[J].畜牧与兽医,2007,39(11):71-74.
[5] Joseph P,Suman S P,Li S,et al. Massspectrometric characterizationand thermostability of turkey myoglobin[J].LWTFood Science and Technology,2010,43(2):273-278.
[6]Berisha A,Yasushi E,Fujimoto K.The effect of heat-induced structural changes on the prooxidant activity of myoglobin[J].Italian Journal of Food Science,2003,15(1):85-94.
[7]王琳可.火候对卤煮鸡腿质构、色泽的影响研究[D].郑州:河南农业大学,2015.
[8]黄明,黄峰,张首玉,等.热处理对猪肉食用品质的影响[J].食品科学,2009,30(23):189-192.
[9]黄甜,严成,黄业传,等.高压结合热处理对猪肉色泽的影响[J].食品工业科技,2015,36(2):85-89.
[10]李升升.热处理对牦牛肉品质的影响及其相关性分析[J].食品与机械,2016(4):207-210.
[11]张淼,王道营,张牧焓,等.一磷酸腺苷与肌动球蛋白相互作用的荧光光谱研究[J].食品科学,2016,37(17):32-37.
[12]张玉姣,唐乾,曹洪玉,等.酸诱导拥挤条件下肌红蛋白及突变体去折叠过程[J].物理化学学报,2013,29(8):1785-1792.
[13]Schmal H,Niemeyer P,Zwingmann J,et al.Relationships of hydrophobicity and net charge to the solubility of milk and soy proteins[J].Journal of Food Science,1985,50(2):486-491.
[14]吴丹,徐桂英.光谱法研究蛋白质与表面活性剂的相互作用[J].物理化学学报,2006,22(2):254-260.
[15]王斌.中药小分子与牛血清白蛋白的相互作用[D].武汉:中南民族大学,2011:39-41.
[16]张莹莹,曹洪玉,唐乾,等.不同类型表面活性剂与高铁肌红蛋白相互作用[J].物理化学学报,2011,27(12):2907-2914.
[17]Ladokhin A S. Fluorescence spectroscopy in peptide and protein analysis[M]. New York:Encyclopedia of Analytical Chemistry,John Wiley&Sons,2006:5762-5779.
[18]吴名草,金邦荃,陈许明,等.紫外照射下高铁肌红蛋白变化的同步荧光光谱指认[J].激光与光电子学进展,2013,50(5)197-203.
[19]王玮,葛毅强,陈颖,等.超高压对高铁肌红蛋白还原酶活性及二级结构的影响[J].中国食品学报,2015,15(10):134-140.
[20]程兵兵.hPRS1及其突变体圆二色光谱研究[D].合肥:中国科学技术大学,2007.
[21]Taheri-Kafrani A,Asgari-Mobarakeh E,Bordbar A K,et al. Structure-function relationship ofβ-lactoglobulin in the presenceof dodecyltrimethyl ammonium bromide[J]. Colloids and Surfaces B Biointerfaces,2010,75(1):268-274.
[22]Moriyama Y,Takeda K. Critical temperature of secondary structural change of myoglobin in thermal denaturation up to 130degrees C and effect of sodium dodecyl sulfate on the change[J].Journal of Physical Chemistry B,2010,114(7):2430.
[23]曹莹莹.超高压结合热处理对肌球蛋白凝胶特性的影响研究[D].南京:南京农业大学,2012.
[24]温斯颖.不同种类动物肌肉蛋白质消化产物比较研究[D].南京:南京农业大学,2015.