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投稿时间：2025-01-27
投稿时间：2025-01-27
中文摘要: 该研究以金枪鱼鱼卵为原料，以鱼卵中卵黄高磷蛋白得率为评价指标，通过筛选蛋白酶以及单因素和响应面试验，探究加酶量、酶解时间、料液比对卵黄高磷蛋白得率的影响，并对最佳工艺下制得的卵黄高磷蛋白进行了十二烷基硫酸钠-聚苯烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis，SDS-PAGE）、等电点、紫外光谱、氮磷比、钙结合能力等指标分析。结果发现，卵黄高磷蛋白的最优制备工艺选用中性蛋白酶为水解酶，工艺条件为pH7、酶解温度55 ℃、加酶量3 000 U∕g、酶解时间1.5 h、料液比1∶21（g∕mL）。在此条件下，卵黄高磷蛋白得率达到6.02 mg∕g，氮磷比6.18，等电点3.5；SDS-PAGE显示卵黄高磷蛋白分子量主要分布在32、37 kDa附近，氨基酸组成分析表明卵黄高磷蛋白中含有丰富的酸性氨基酸和丝氨酸，钙结合能力为9.25 mg∕g。
中文关键词: 金枪鱼卵 卵黄高磷蛋白 酶解工艺 钙结合能力 Box-Behnken 响应面法
Abstract:By using tuna roe as raw material and phosvitin （Pv） yield as the evaluation index，the effects of enzyme dosage，hydrolysis time，and solid-liquid ratio on the yield of Pv were investigated through protease screening，single-factor experiments，and response surface methodology.The characteristics of tuna Pv prepared by the optimal process were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE），isoelectric point，Ultraviolet spectrum，nitrogen-to-phosphorus ratio （N∕P ratio），and calcium binding capacity.The results showed that the optimal preparation process of Pv used neutral protease as the hydrolase.Under the conditions of pH at 7，hydrolysis temperature at 55 ℃，enzyme dosage of 3 000 U∕g，hydrolysis time of 1.5 h，and solid-liquid ratio of 1∶21 （g∕mL），the yield of Pv reached 6.02 mg∕g.Its nitrogen-tophosphorus molar ratio was 6.18，and the isoelectric point was 3.5.SDS-PAGE results showed that Pv bands were mainly distributed around 32 kDa and 37 kDa.Amino acid composition analysis indicated that Pv was rich in acidic amino acids and serine，and the calcium binding capacity of Pv was 9.25 mg∕g.
keywords: tuna roe phosvitin enzymatic hydrolysis process calcium binding capacity Box-Behnken response surface methodology
文章编号：202609010 中图分类号： 文献标志码：
基金项目:舟山科技计划项目（2024C03004）
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