基于HS-GC-IMS和HS-SPME-GC-MS的蛋白酶对豆粕挥发性风味的影响分析

Analysis of Effects of Proteases on Volatile Flavor of Soybean Meal Based on HS-GC-IMS and HS-SPME-GC-MS

为研究不同蛋白酶酶解对豆粕挥发性风味成分的影响,选用4种蛋白酶(碱性蛋白酶、中性蛋白酶、木瓜蛋白酶、风味蛋白酶)对豆粕进行酶解,采用顶空-气相色谱-离子迁移谱(Headspace-gas chromatography-ion mobility spectroscopy,HS-GC-IMS)和顶空固相微萃取-气相色谱-质谱(Headspace solid phase microextraction-gas chromatography-mass spectrometry,HS-SPME-GC-MS)联用技术分析不同豆粕酶解物(Soybean meal hydrolysates,SMH)的挥发性风味成分,并结合主成分分析(Principal component analysis,PCA)、热图聚类和正交偏最小二乘判别法(Orthogonal partial least squares-discriminant analysis,OPLS-DA)对不同SMH进行分析。结果表明:碱性蛋白酶、中性蛋白酶、木瓜蛋白酶和风味蛋白酶酶解豆粕的挥发性风味成分存在较大差异。HS-GC-IMS鉴定出84种挥发性成分,筛选得到33种差异风味物质,发现酶解后酮类物质显著降低而醛类、醇类和酯类物质含量明显增加。PCA结果表明不同SMH之间的风味存在显著差异。最终通过OPLS-DA筛选出贡献较大的挥发性化合物,同时构建出可靠的用以鉴别SMH的模型。HS-SPME-GC-MS检测出103种差异风味物质,可用于区分不同SMH,被检出的挥发性组分中醛类、醇类和酮类等化合物为SMH风味的形成做出主要贡献,明晰了部分风味化合物形成的原因。PCA和聚类热图结果表明不同蛋白酶酶解对豆粕的挥发性风味物质的种类和含量有显著影响,其中,风味蛋白酶和木瓜蛋白酶对豆粕的风味改善最为显著。

To investigate the effect of enzymatic digestion of different proteases on the volatile flavor components of soybean meal,four proteases(alkaline protease,neutral protease,papain,flavor protease) were selected to enzyme soybean meal,using a headspace-gas chromatography-ion mobility spectroscopy(HS-GC-IMS) and headspace solid phase microextraction-gas chromatography-mass spectrometry(HS-SPME-GC-MS) to analyze volatile flavor composition of different soybean meal hydrolysates(SMH),combined with the principal component analysis(PCA),heatmap clustering,and orthogonal partial least squares-discriminant analysis(OPLS-DA) to analyze the different SMH.The results showed that the volatile flavor components of alkaline protease,neutral protease,papain and flavor protease were quite different.HS-GC-IMS identified 84 volatile components,and screened 33 differential flavor substances,which found that the content of ketones and aldehyde,alcohols and esters were increased significantly after enzymatic digestion.The PCA results indicated significant differences in flavor between different SMH.Finally,volatile compounds with large contributions were selected by OPLS-DA,and reliable models used to identify SMH were constructed.HS-SPME-GC-MS detected 103 different flavor substances that could be used to distinguish different SMH.The volatile compounds such as aldehyde,alcohols,and ketones made major contributions to the formation of SMH flavor,which clarified the reasons for the formation of some flavor compounds.PCA and cluster heat map results showed that the enzyme digestion of different proteases had a significant influence on the type and content of volatile flavor substances of soybean meal,among which,flavor protease and papain showed the most significant flavor improvement of soybean meal.