提取时间对鸡骨蛋白凝胶特性和蛋白二级结构的影响

doi:10.3864/j.issn.0578-1752.2017.05.013

摘要/Abstract

摘要： 【目的】研究提取时间对鸡骨蛋白（chicken bone protein，CBP）凝胶特性的影响，为利用鸡骨副产物制备食用凝胶提供参考。【方法】采用热压抽提法提取CBP，于4℃孵育制备CBP凝胶，测定不同抽提时间的抽提液中总固形物、粗蛋白、羟脯氨酸含量、蛋白质二级结构、分子量分布的变化，探讨抽提时间对CBP凝胶色差、凝胶强度的影响，并分析各指标之间的相关性。【结果】抽提时间显著影响鸡骨蛋白的提取率（P＜0.05），导致蛋白结构发生显著变化进而影响其凝胶特性。抽提时间从0—120 min，抽提液中总固形物、粗蛋白和羟脯氨酸含量均显著增加（P＜0.05）；随着提取时间延长（40—120 min），蛋白发生显著降解，分子量10—30 kDa的肽段从40 min的59.82%降低到120 min的13.99%，分子量＜10 kDa的肽段从40 min的35.46% 升高到120 min的86.01%；提取时间延长引起蛋白发生部分程度变性，导致其二级结构改变，0—40 min时的CBP酰胺Ⅰ带由100%的α-螺旋组成，60—90 min时出现9.9%—17.6%的β-折叠结构，120 min时β-折叠全部降解；对CBP凝胶特性的分析表明，抽提时间为0 min时，鸡骨蛋白不形成凝胶，而抽提20、40和60 min时的凝胶强度无显著性差异，均优于90、120 min组；随着提取时间延长（20—120 min），提取液的透明度显著增加（P＜0.05），但40和60 min组的CBP制备的凝胶色泽最佳（P＜0.05）；抽提时间（20—120 min）对凝胶持水性无显著影响（P＞0.05）。相关性分析表明，CBP中羟脯氨酸含量、CBP降解程度与凝胶特性显著相关（P＜0.05）。【结论】热压抽提时间对CBP凝胶特性有显著影响，综合考虑蛋白得率、凝胶色泽、强度等因素，40和60 min提取的鸡骨蛋白较适合制备凝胶。

关键词: 鸡骨蛋白, 热压抽提法, 提取时间, 凝胶特性, 二级结构

Abstract: 【Objective】Hot-pressure extraction was utilized to prepare chicken bone protein (CBP) to study the effect of extraction time on gelatin properties. The study was expected to provide a reference for the preparation technology of edible gelatin by using chicken bone by-products. 【Method】After being adjusted to the same concentration of protein, CBP at different extraction times were incubated at 4℃ to form gelatin. The content of total soluble substance (TSS), crude protein and hydroxyproline (Hyp), secondary structure of protein and distribution of molecular weight (MW), as well as color of gelatin and gel strength were studied. Furthermore, correlation analysis between indexes of CBP was also performed to establish possible linkages between physicochemical properties and gelatin properties of CBP.【Result】The results showed that hot-pressure extraction time had a significant effect on the yield of CBP (P＜0.05), leading to the variable structure of protein and consequently affected the properties of gelatin made by CBP. The contents of TSS, crude protein and Hyp were increased significantly during extraction (0-120 min) (P＜0.05). The result of MW distribution suggested that protein experienced dramatic degradation from 40 to 120 min (P＜0.05). The proportion of MW between 10 and 30 kDa decreased from 59.82% (40 min) to 13.99% (120 min) while the ratio of MW ＜10 kDa increased from 35.46% (40 min) to 86.01% (120 min). In addition, the extraction time caused the denaturation of protein and thus leading to the change of secondary structure. The CBP was composed of 100 percent of α-helix (0-40 min), and then β-sheet emerged at 60 and 90 min (9.9% and 17.6%, respectively) but disappeared at 120 min, which indicated that the secondary structure of protein experienced transformation during extraction. Results of CBP gelatin properties suggested that CBP at 0 min could not form a gelatin and thus had no gelatin properties. Gel strength at 20, 40 and 60 min was at same level and better than CBP gelatins of 90 and 120 min. In addition, transparency of CBE solutions was significantly impacted by extraction time, which increased sharply during hot-pressure extraction (20-120 min) (P＜0.05). Meanwhile, extraction time significantly affected the color of gelatins (P＜0.05) and gelatin at 40 and 60 min met the requirements of high lightness, low yellowness and high whiteness. However, water holding capacity was insensitive to the extraction time (P＞0.05). Correlation analysis showed that the content of Hyp, degradation of protein were considerably related to gelatin properties (P＜0.05).【Conclusion】In conclusion, extraction time could significantly affect the gelatin properties prepared by CBP at different extraction times. Given yield of protein, color of gelatin, gel strength, etc., CBP extracted at 40 min and 60 min were better proper for preparation of gelatin.

Key words: chicken bone protein, hot-pressure extraction, extraction time, gelatin properties, secondary structure

岳鉴颖，王金枝，张春晖，杜桂红，许雄. 提取时间对鸡骨蛋白凝胶特性和蛋白二级结构的影响[J]. 中国农业科学, 2017, 50(5): 903-912.

YUE JianYing, WANG JinZhi, ZHANG ChunHui, DU GuiHong, XU Xiong. Effect of Extraction Time on Characteristics of Gelatin and Secondary Structure of Chicken Bone Protein[J]. Scientia Agricultura Sinica, 2017, 50(5): 903-912.

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