Controlling cis/trans isomerism of monounsaturated fatty acids via a recombinant cytochrome c-type cis/trans fatty acid isomerase

Park, J.-Y., Jung, Y.-S., Charalampopoulos, D. ORCID: https://orcid.org/0000-0003-1269-8402, Park, K.-M. ORCID: https://orcid.org/0000-0002-4706-8681 and Chang, P.-S. ORCID: https://orcid.org/0000-0001-9645-7010 (2024) Controlling cis/trans isomerism of monounsaturated fatty acids via a recombinant cytochrome c-type cis/trans fatty acid isomerase. Food Control, 160. 110319. ISSN 1873-7129 doi: 10.1016/j.foodcont.2024.110319

Abstract/Summary

Cytochrome c-type cis/trans fatty acid isomerase (CTI) has been proposed to control cis/trans isomerism of unsaturated fats in lipid-related food products. A gene encoding wildtype CTI from Pseudomonas putida KT2440 was introduced into the pET26b/pEC86 co-expression system, and the heme C cofactor was covalently bound into the expressed CTI protein through in vivo cytochrome c maturation. The recombinant CTI, purified from Escherichia coli BL21(DE3), catalyzed the cis/trans isomerization of three edible monounsaturated fatty acids. It exhibited strong substrate selectivity for palmitoleic acid (C16:1, cis-Δ9), reaching an 80.93 ± 1.78% conversion at reaction equilibrium. Notably, its promiscuity for other fatty acids (oleic acid: 29.21 ± 5.01% and cis-vaccenic acid: 51.21 ± 0.05%) was observed. Under the optimum reaction conditions (pH 7.5 and 15 °C), the kinetic parameters (Vmax, Km, and kcat) of CTI were derived as 0.035 mM·min−1, 0.267 mM, and 0.141 sec−1, respectively, and the final catalytic efficiency (kcat/Km) was calculated as 5.26 × 102 M−1·sec−1. Furthermore, structural properties of CTI were analyzed using deep learning-based protein structure prediction, suggesting the potential for specificity variability by altering loop dynamics and helix interactions surrounding the heme-binding motif. The following results would provide theoretical and practical information for CTI enzymes as novel promising industrial catalysts to control cis/trans isomerism of lipids in food products.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/114807
Identification Number/DOI	10.1016/j.foodcont.2024.110319
Refereed	Yes
Divisions	Life Sciences > School of Chemistry, Food and Pharmacy > Department of Food and Nutritional Sciences > Food Research Group
Publisher	Elsevier
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