假交替单胞菌κ-卡拉胶酶的酶解工艺优化及产物分析  

Optimization of the Enzymatic Hydrolysis Process Using Pseudoalteromonas sp.κ-Carrageenase and Analysis of the Hydrolysates

作　　者：李鹤宾 牟明静 洪璇 吴婷 陈艳红[2] 姜泽东[2] 朱艳冰[2] LI Hebin;MOU Mingjing;HONG Xuan;WU Ting;CHEN Yanhong;JIANG Zedong;ZHU Yanbing(Department of Pharmacy,Xiamen Medical College,Xiamen 361023,China;College of Ocean Food and Biological Engineering,Jimei University,Xiamen 361021,China)

机构地区：[1]厦门医学院药学系,福建厦门361023 [2]集美大学海洋食品与生物工程学院,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2022年第5期417-423,共7页Journal of Jimei University：Natural Science

基　　金：福建省自然科学基金项目(2020J01679);漳州市科技重大专项(ZZ2019ZD15)。

摘　　要：为探索假交替单胞菌κ-卡拉胶酶的酶解工艺,以κ-卡拉胶为底物,还原糖生成量为评价指标,对酶解工艺进行优化。采用3,5-二硝基水杨酸法和苯酚-硫酸法分别测定还原糖和总糖含量,计算酶解产物的平均聚合度,并利用质谱鉴定酶解产物。结果显示,假交替单胞菌κ-卡拉胶酶降解κ-卡拉胶的优化工艺条件为:加酶量为0.35 U(反应体系5 mL),反应温度为40℃,反应pH=8.0,κ-卡拉胶底物质量浓度为9 g/L。在此条件下,酶解反应240 min后产生的还原糖质量浓度为1.531 g/L,酶解产物的平均聚合度为2。该κ-卡拉胶酶酶解反应的K=2.07 g/L,V=7.25 U/mg。质谱分析显示,假交替单胞菌κ-卡拉胶酶降解κ-卡拉胶的产物为κ-卡拉胶二糖和κ-卡拉胶四糖。In order to explore the enzymatic hydrolysis process using κ-carrageenase from Pseudoalteromonas sp.,κ-carrageenan was used as the substrate and the amount of reducing sugar production was used as the evaluation index to optimize the enzymatic hydrolysis process.The contents of the reducing sugar and total sugar were determined by the methods of 3,5-dinitrogen salicylic acid and phenol-sulfuric acid, respectively.The average polymerization degree of the enzymatic hydrolysates was calculated, and the hydrolysis products were analyzed by mass spectrometry.The results showed that the optimal process for the κ-carrageenan degradation by Pseudoalteromonas sp.κ-carrageenase were as follows: enzyme amount of 0.35 U(5 mL reaction system),reaction temperature of 40 ℃,reaction pH value of 8.0,and κ-carrageenan substrate concentration of 9 g/L.Under the conditions, the concentration of the reducing sugar produced was 1.531 g/L,and the average polymerization degree of the enzymatic hydrolysates was 2 after reaction for 240 min.The Kvalue of the κ-carrageenase was 2.07 g/L,and the Vvalue was 7.25 U/mg.Mass spectrometry analysis showed that the products of κ-carrageenan degradation by Pseudoalteromonas sp.κ-carrageenase were κ-carrageenan disaccharide and κ-carrageenan tetraose.

关 键 词：Κ-卡拉胶 酶解 工艺优化 聚合度 κ-卡拉胶寡糖 

分 类 号：TQ925[轻工技术与工程—发酵工程]