An improved method for rapid evaluation of enzymatic cis/trans isomerization of C18:1 monounsaturated fatty acids

Jun-Young Park; Hyeon-Whoong Choi; Kyung-Min Park; Pahn-Shick Chang

doi:10.1016/j.foodchem.2022.134618

An improved method for rapid evaluation of enzymatic cis/trans isomerization of C18:1 monounsaturated fatty acids

Food Chem. 2023 Mar 15;404(Pt A):134618. doi: 10.1016/j.foodchem.2022.134618. Epub 2022 Oct 17.

Authors

Jun-Young Park¹, Hyeon-Whoong Choi², Kyung-Min Park³, Pahn-Shick Chang⁴

Affiliations

¹ Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea.
² Department of Food Science and Biotechnology, Wonkwang University, Iksan 54538, Republic of Korea.
³ Department of Food Science and Biotechnology, Wonkwang University, Iksan 54538, Republic of Korea. Electronic address: kmpark79@wku.ac.kr.
⁴ Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea; Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea; Center for Agricultural Microorganism and Enzyme, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: pschang@snu.ac.kr.

PMID: 36444044
DOI: 10.1016/j.foodchem.2022.134618

Abstract

Cytochrome c-type cis/trans fatty acid isomerase (CTI) is a promising candidate for directly controlling cis/trans fatty acid isomerism in lipids-related food products like partially hydrogenated vegetable oils. In this study, to establish a sophisticated analysis platform for the CTI assay, we constructed the reversed micelle reaction system and improved the processes of methylation and GC-FID analysis of C_18:1cis/trans monounsaturated fatty acid (MUFA) isomers. Highly stable AOT/isooctane reversed micelles were formed in the presence of periplasmic fractions of Pseudomonas putida KT2440. Using a mid-content cyanopropyl phase DB-FastFAME column, C_18:1cis/trans-MUFAs were analyzed rapidly and resolved with resolution factors over 1.34. Based on the newly established assay, the catalytic activity of the periplasmic fraction was precisely determined, and its kinetic parameters (V_max and K_m) were derived as 0.021 mM·min^-1 and 0.68 mM, respectively. The following results can provide practical information for investigating CTIs in the fields of food and lipid chemistry.

Keywords: Cis/trans fatty acid isomerase; Cis/trans unsaturated fatty acid; DB-FastFAME; Elaidic acid, PubChem CID: 637517; Fatty acid methyl ester; Gas chromatography; Methyl cis-vaccenate, PubChem CID: 536405; Methyl elaidate, PubChem CID: 5280590; Methyl oleate, PubChem CID: 5364509; Methyl trans-vaccenate, PubChem CID: 5364432; Oleic acid, PubChem CID: 445639; Reversed micelle; cis-Vaccenic acid, PubChem CID: 5282761; trans-Vaccenic acid, PubChem CID: 5281127.

MeSH terms

Biological Assay
Fatty Acids
Fatty Acids, Monounsaturated*
Isomerism
Micelles
Trans Fatty Acids*

Substances

Fatty Acids, Monounsaturated
Trans Fatty Acids
Fatty Acids
Micelles