基于两步水解法制备三疣梭子蟹调味品的酶解工艺优化研究

doi:10.3785/j.issn.1008-9209.2021.04.282

浙江大学学报（农业与生命科学版）

2022, Vol. 48

Issue (3): 321-335 DOI: 10.3785/j.issn.1008-9209.2021.04.282

食品科学

基于两步水解法制备三疣梭子蟹调味品的酶解工艺优化研究

陈瑜^1,^2,³(

),马剑锋⁴,许丹^1,^2,³,张小军^1,^2,³(

),何鹏飞^1,^2,³

^1.浙江省海洋水产研究所，浙江舟山 316021
^2.农业农村部重点渔场渔业资源科学观测实验站，浙江舟山 316021
^3.浙江省海洋渔业资源可持续利用技术研究重点实验室，浙江舟山 316021
^4.浙江海洋大学食品与医药学院，浙江舟山 316022

Optimization of enzymolysis technology for preparing Portunus trituberculatus seasoning based on a two-step hydrolysis method

Yu CHEN^1,^2,³(

),Jianfeng MA⁴,Dan XU^1,^2,³,Xiaojun ZHANG^1,^2,³(

),Pengfei HE^1,^2,³

^1.Zhejiang Marine Fisheries Research Institute, Zhoushan 316021, Zhejiang, China
^2.Scientific Observing and Experimental Station of Fishery Resources for Key Fishing Grounds, Ministry of Agriculture and Rural Affairs, Zhoushan 316021, Zhejiang, China
^3.Key Laboratory of Sustainable Utilization of Technology Research for Fishery Resource of Zhejiang Province, Zhoushan 316021, Zhejiang, China
^4.Food and Medicine School, Zhejiang Ocean University, Zhoushan 316022, Zhejiang, China

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摘要：

本研究旨在建立以三疣梭子蟹蒸煮液为原料制备调味品的最佳酶解工艺。以鲜味氨基酸含量为评价指标，通过响应面实验设计优化确定碱性蛋白酶、风味蛋白酶依次水解的两步酶解工艺，并采用顶空固相微萃取（headspace-solid phase microextraction, HS-SPME）结合气相色谱-质谱联用（gas chromatography-mass spectrometry, GC-MS）技术对三疣梭子蟹二次酶解后产生的挥发性风味物质进行定性与定量分析。结果表明：一次酶解最佳工艺条件为碱性蛋白酶添加量6 825 U/g、初始水解pH 8.45、水解时间2.50 h，得到的酶解液中鲜味氨基酸质量浓度为（7.18±0.09） mg/mL；二次酶解最佳酶解工艺条件为风味蛋白酶添加量3 750 U/g、初始水解pH 6.81、水解时间3.33 h，得到的酶解液中鲜味氨基酸质量浓度为（10.49±0.12） mg/mL，两步酶解优化后酶解液中鲜味氨基酸含量比梭子蟹蒸煮液提高了135.2%。二次酶解液中共定性检测到62种挥发性风味成分，其中己醛、苯甲醛和不饱和醇为挥发性风味主要贡献物质。本研究结果为研发蒸煮液调味产品和三疣梭子蟹加工副产物高值化利用提供了理论依据。

关键词： 三疣梭子蟹蒸煮液; 两步水解; 鲜味氨基酸; 响应面分析; 挥发性风味物质

Abstract:

The objective of this study was to establish the optimum enzymolysis technology for Portunus trituberculatus seasoning based on the cooking liquor. Taking the flavor amino acid contents as evaluation indicators, a two-step enzymolysis technology with alcalase and flavorzyme was determined by response surface optimization methodology, and volatile flavor compounds in the secondary enzymolysis liquor were qualitatively and quantitatively analyzed by headspace-solid phase microextraction (HS-SPME) with gas chromatography-mass spectrometry (GC-MS). The results indicated that the optimum technological conditions of the primary enzymolysis were alcalase of 6 825 U/g, primary hydrolysis pH 8.45, hydrolysis time of 2.50 h, and the content of flavor amino acid in the hydrolysate was (7.18±0.09) mg/mL. The optimum technological conditions of the secondary enzymolysis were flavorzyme of 3 750 U/g, primary hydrolysis pH 6.81, hydrolysis time of 3.33 h, and the content of flavor amino acid in flavor precursors of P. trituberculatus seasoning was (10.49±0.12) mg/mL, which was 135.2% higher than that of P. trituberculatus cooking liquor. There were 62 volatile flavor compounds in the secondary hydrolysate; among them, hexanal, benzaldehyde and unsaturated alcohol were the main flavor components.The results provide a theoretical basis for the research of seasoning products of cooking liquor and high value utilization of P. trituberculatus.

Key words: Portunus trituberculatus cooking liquor two-step hydrolysis flavor amino acids response surface analysis volatile flavor compounds

收稿日期: 2021-04-28 出版日期: 2022-07-07

CLC:

TS 254.9

基金资助: 浙江省舟山市科技计划项目(2019C31051)

通讯作者: 张小军 E-mail: chenxiaoyu2141@163.com;xiaojun3627@163.com

作者简介: 陈瑜（https://orcid.org/0000-0001-5053-9766），E-mail：chenxiaoyu2141@163.com

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引用本文:

陈瑜,马剑锋,许丹,张小军,何鹏飞. 基于两步水解法制备三疣梭子蟹调味品的酶解工艺优化研究[J]. 浙江大学学报（农业与生命科学版）, 2022, 48(3): 321-335.

Yu CHEN,Jianfeng MA,Dan XU,Xiaojun ZHANG,Pengfei HE. Optimization of enzymolysis technology for preparing Portunus trituberculatus seasoning based on a two-step hydrolysis method. Journal of Zhejiang University (Agriculture and Life Sciences), 2022, 48(3): 321-335.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2021.04.282 或 https://www.zjujournals.com/agr/CN/Y2022/V48/I3/321

表1 各种蛋白酶的酶解反应特性

表2 复合蛋白酶的酶解条件

表3 感官评分标准

表4 碱性蛋白酶响应面法优化实验因素与水平

表5 风味蛋白酶响应面法优化实验因素与水平

图1 蛋白酶种类对游离氨基酸含量（A）和感官评分（B）的影响图中序号所代表的含义详见表1。短栅上不同小写字母表示在P＜0.05水平差异有统计学意义。

图2 蛋白酶添加量对游离氨基酸含量的影响

图3 初始水解pH对游离氨基酸含量的影响

图4 水解时间对游离氨基酸含量的影响

表6 一次酶解响应面实验设计方案及结果

表7 二次酶解响应面实验设计方案及结果

表8 一次酶解优化实验方差分析

表9 二次酶解优化实验方差分析

图5 一次酶解实验各因素交互作用对鲜味氨基酸含量影响的曲面图

图6 二次酶解实验各因素交互作用对鲜味氨基酸含量影响的曲面图

表10 三疣梭子蟹蒸煮液与两步酶解优化酶解液中游离氨基酸含量 (mg/mL)

表11 酶解液中的挥发性风味物质

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