Extraction Optimization and Analysis on Properties of Protein on Peony Seed Meal
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摘要: 本研究通过碱溶酸沉法对牡丹籽粕中的蛋白进行提取。选取料液比、pH、时间和温度进行单因素研究，结合响应面法优化获得最佳提取工艺，并对蛋白的物化特性进行分析。确定最佳工艺条件：料液比1:25 g/mL，pH10.6，温度55 ℃，时间130 min时，蛋白得率为23.81%±0.04%。在此条件下获得的牡丹籽粕蛋白中含有18种氨基酸；蛋白的持水性和持油性分别为3.72和3.67 g/g；起泡性和泡沫稳定性在pH2~4时均明显降低，pH4时最小，pH6~10之间时，起泡性持续增加，泡沫稳定性明显上升后略有下降；乳化性和乳化稳定性随pH增大而增加，与粒径和Zeta电位所反映的结果相符。本研究为牡丹籽粕蛋白的工业化生产和综合利用提供了理论依据。Abstract: In this study, an alkali solution and acid precipitation method were used to extract protein from peony seed meal. Solid-liquid ratio, pH, time, and temperature were selected to carry out a single-factor study and combined with the response surface method to optimize the optimal extraction process. The optimum process conditions were determined: The protein yield reached 23.81%±0.04% when the ratio of solid to liquid was 1:25 g/mL, pH10.6, the temperature was 55 ℃, and time was 130 min. The peony seed meal protein obtained under these conditions contained 18 kinds of amino acids. The water and oil retention of peony seed meal protein was 3.72 g/g and 3.67 g/g, respectively. Foamability and foam stability decreased significantly at pH2~4, and the lowest was at pH4. Between pH6 and 10, foamability continued to increase, while foam stability increased significantly and then decreased slightly. With the increase in pH, the emulsification and emulsification stability increased. The results were consistent with the variation trend of particle size and Zeta potential. This study provided a theoretical basis for the industrial production and comprehensive utilization of peony seed meal protein.
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图 5 温度对牡丹籽粕蛋白得率的影响
Figure 5. Effect of temperature on protein yield of peony seed meal
图 6 各因素交互作用对牡丹籽粕蛋白得率影响的响应面图
Figure 6. Response surface of various factors on protein yield of peony seed meal
图 7 牡丹籽粕蛋白的持水性和持油性
Figure 7. The water holding capacity and oil holding capacity of peony seed meal protein
图 8 不同pH条件下牡丹籽粕蛋白溶液的粒径
Figure 8. Particle size of peony seed meal protein solution under different pH conditions
图 9 不同pH条件下牡丹籽粕蛋白溶液的Zeta电位
Figure 9. Zeta potential of peony seed meal protein solution under different pH conditions
图 10 不同pH条件下牡丹籽粕蛋白的起泡性及泡沫稳定性
Figure 10. Foaming ability and foam stability of peony seed meal protein under different pH
图 11 不同pH条件下牡丹籽粕蛋白的乳化性及乳化稳定性
Figure 11. Emulsification and emulsifying stability of peony seed meal protein under different pH
表 1 单因素变量实验水平设计
Table 1. Factor level of single factor variable design
实验设计 料液比（g/mL） pH 温度（℃） 时间（min） 1 1:15、1:20、1:25、1:30、1:35 10 40 30 2 1:25 8、9、10、11、12 40 30 3 1:25 11 40 70、90、110、130、150 4 1:25 11 40、45、50、55、60 130 
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表 2 响应面试验分析因素与水平
Table 2. Response surface test analysis factors and levels
水平 因素 料液比（g/mL） pH 温度（℃） −1 1:20 10 50 0 1:25 11 55 1 1:30 12 60
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表 3 响应面试验结果
Table 3. Response surface test results
试验号 A 料液比 B pH C 温度 Y 得率（%） 1 −1 0 1 22.45 2 −1 1 0 17.73 3 −1 0 −1 22.04 4 −1 −1 0 22.29 5 0 0 0 23.38 6 0 1 −1 19.89 7 0 −1 1 23.06 8 0 0 0 23.78 9 0 0 0 23.61 10 0 0 0 23.71 11 0 1 1 19.48 12 0 −1 −1 23.18 13 0 0 0 23.56 14 1 1 0 19.6 15 1 −1 0 22.37 16 1 0 −1 22.87 17 1 0 1 22.57
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表 4 回归模型方差分析
Table 4. Regression model variance analysis
来源 平方和 自由度 均方 F值 P值 差异性 模型 51.40 9 5.71 134.63 <0.0001 ** A 料液比 1.05 1 1.05 24.78 0.0016 ** B pH 25.20 1 25.20 594.20 <0.0001 ** C 温度 0.0221 1 0.0221 0.5198 0.4943 AB 0.8010 1 0.8010 18.88 0.0034 ** AC 0.1260 1 0.1260 2.97 0.1284 BC 0.0210 1 0.0210 0.4957 0.5042 A2 4.34 1 4.34 102.31 <0.0001 ** B2 18.48 1 18.48 435.76 <0.0001 ** C2 0.0512 1 0.0512 1.21 0.3084 残差 0.2969 7 0.0424 失拟项 0.2027 3 0.0676 2.87 0.1677 不显著 绝对误差 0.0943 4 0.0236 总和 51.70 16 注：**表示差异极显著（P<0.01）；*表示差异显著（P<0.05）。
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表 5 牡丹籽粕蛋白的氨基酸组成及含量
Table 5. Amino acid composition and content of peony seed meal protein
氨基酸 缩写 样品含量（%） 天冬氨酸 Asp 9.00±0.26 谷氨酸 Glu 22.79±0.30 丝氨酸 Ser 4.39±0.04 甘氨酸 Gly 4.16±0.01 组氨酸 His 1.77±0.03 精氨酸 Arg 6.78±0.13 苏氨酸* Thr 2.45±0.04 丙氨酸 Ala 3.91±0.00 脯氨酸 Pro 3.86±0.18 酪氨酸 Tyr 2.55±0.06 缬氨酸* Val 4.87±0.06 蛋氨酸* Met 1.06±0.04 胱氨酸 Cyr 1.16±0.04 异亮氨酸* Ile 3.53±0.08 亮氨酸* Leu 6.53±0.07 苯丙氨酸* Phe 3.19±0.11 赖氨酸* Lys 1.79±0.08 色氨酸* Trp 0.63±0.01 总量 84.42 注：*表示必需氨基酸。
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