Stability comparison of four lipases and catalytic mechanism during the synthesis of 1,3‐di‐oleic‐2‐medium chain triacylglycerols in a trace water‐in‐oil system: Experimental analyses and computational simulations

Journal of Food BiochemistryVolume 45, Issue 5 e13667 FULL ARTICLE Stability comparison of four lipases and catalytic mechanism during the synthesis of 1,3-di-oleic-2-medium chain triacylglycerols in a trace water-in-oil system: Experimental analyses and computational simulations Bin Peng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: Data curation, Writing - original draftSearch for more papers by this authorTing Luo, Corresponding Author Ting.Luo@utsouthwestern.edu orcid.org/0000-0002-3394-7861 State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Correspondence Ting Luo and Ze-Yuan Deng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi 330047, China. Email: zeyuandeng@hotmail.com (Z.-Y. D.) and Ting.Luo@utsouthwestern.edu (T. L.) Contribution: Writing - review & editingSearch for more papers by this authorFang Chen, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China School of Public Health, Nanchang University, Nanchang, China Contribution: Formal analysis, Writing - review & editingSearch for more papers by this authorMei Wang, The State Centre of Quality Supervision and Inspection for Camellia Products, Ganzhou, China Contribution: Formal analysis, SoftwareSearch for more papers by this authorJin-Heng Fu, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: Resources, VisualizationSearch for more papers by this authorLiu-Feng Zheng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: Data curation, Methodology, ValidationSearch for more papers by this authorJing Li, orcid.org/0000-0002-2271-7761 State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: ​Investigation, Project administrationSearch for more papers by this authorZe-Yuan Deng, Corresponding Author zeyuandeng@hotmail.com orcid.org/0000-0001-5650-1507 State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Correspondence Ting Luo and Ze-Yuan Deng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi 330047, China. Email: zeyuandeng@hotmail.com (Z.-Y. D.) and Ting.Luo@utsouthwestern.edu (T. L.) Contribution: Funding acquisition, Visualization, Writing - review & editingSearch for more papers by this author Bin Peng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: Data curation, Writing - original draftSearch for more papers by this authorTing Luo, Corresponding Author Ting.Luo@utsouthwestern.edu orcid.org/0000-0002-3394-7861 State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Correspondence Ting Luo and Ze-Yuan Deng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi 330047, China. Email: zeyuandeng@hotmail.com (Z.-Y. D.) and Ting.Luo@utsouthwestern.edu (T. L.) Contribution: Writing - review & editingSearch for more papers by this authorFang Chen, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China School of Public Health, Nanchang University, Nanchang, China Contribution: Formal analysis, Writing - review & editingSearch for more papers by this authorMei Wang, The State Centre of Quality Supervision and Inspection for Camellia Products, Ganzhou, China Contribution: Formal analysis, SoftwareSearch for more papers by this authorJin-Heng Fu, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: Resources, VisualizationSearch for more papers by this authorLiu-Feng Zheng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: Data curation, Methodology, ValidationSearch for more papers by this authorJing Li, orcid.org/0000-0002-2271-7761 State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Contribution: ​Investigation, Project administrationSearch for more papers by this authorZe-Yuan Deng, Corresponding Author zeyuandeng@hotmail.com orcid.org/0000-0001-5650-1507 State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Correspondence Ting Luo and Ze-Yuan Deng, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi 330047, China. Email: zeyuandeng@hotmail.com (Z.-Y. D.) and Ting.Luo@utsouthwestern.edu (T. L.) Contribution: Funding acquisition, Visualization, Writing - review & editingSearch for more papers by this author First published: 09 April 2021 https://doi.org/10.1111/jfbc.13667Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat Abstract In the present study, a kind of structured lipids, namely 1,3-di-oleic-2-medium chain (OMO) triacylglycerols, were synthesized through lipase-catalyzed reactions using coconut oil and rapeseed acid as materials in a trace water-in-oil system. Experimental analysis and computational simulations were undertaken to compare the stability of four lipases including Lipozyme RMIM, Lipozyme TLIM, Novozym 435, and Aspergillus oryzae immobilized lipase (AOIM), and illustrate catalytic mechanism of Novozym 435 during the synthesis of OMO. Fourier transform infrared and molecular dynamics simulation results demonstrated that a decrease in ordered structure (α-helix and β-sheet) led to a reduction in enzyme activity. Compared with Lipozyme RMIM and Novozym 435, Lipozyme TLIM and AOIM exhibited better stability due to a short-chain lid in TLIM, which covers activity sites, and hydrogen bonds formed between activity center of AOIM and water. Among four lipases, AOIM exhibited best catalytic performance: a OMO yield of 30.7% at 3 hr and a good stability of long term (48 hr). Density functional theory results demonstrated that specifically, during the synthesis of OMO triacylglycerol, the addition of Novozym 435 (derived from Candida antarctica lipase B, CALB) substantially lowered reaction barriers (64.4 KJ/mol with CALB vs. 332.7 KJ/mol with no lipase), aiding in the generation of OMO because of the formations of transitional tetrahedral intermediates. A trace water-in-oil system was a green and efficient alternative for lipase-catalyzed production of OMO, and this study provided crucial insights into the stability/instability and catalytic mechanisms of lipase in the synthesis of structured lipids. Practical applications We compared the stability of Lipozyme RMIM, Lipozyme 435, Lipozyme TLIM, and AOIM during the synthesis of the OMO triacylglycerols in a trace water-in-oil system, where exhibited a high catalytic activity of lipase in water–oil interface. AOIM had the highest stability and showed the best catalytic performance due to the formation of hydrogen bonds. Besides, for the first time, the transition tetrahedral structure was revealed in the enzymatic synthesis of medium- and long-chain triacylglycerols. This study provides a rational approach to compare lipase stability and a possible hint to choose appropriate enzyme in a specific catalytic condition. Open Research DATA AVAILABILITY STATEMENT The data that support the findings of this study are openly available in Figure 1 and Supplementary data Table 2 at https://doi.org/10.1016/j.foodchem.2019.126130 Supporting Information Filename Description jfbc13667-sup-0001-Supinfo.docxWord document, 3.5 MB Supplementary Material Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article. Volume45, Issue5May 2021e13667 RelatedInformation