Engineering a Feruloyl–Coenzyme A Synthase for Bioconversion of Phenylpropanoid Acids into High-Value Aromatic Aldehydes

Abstract

Aromatic aldehydes find extensive applications in food, perfume, pharmaceutical, and chemical industries. However, a limited natural enzyme selectivity has become the bottleneck of bioconversion of aromatic aldehydes from natural phenylpropanoid acids. Here, based on the original structure of feruloyl–coenzyme A (CoA) synthetase (FCS) from Streptomyces sp. V-1, we engineered five substrate-binding domains to match specific phenylpropanoid acids. FcsCIAE407A/K483L, FcsMAE407R/I481R/K483R, FcsHAE407K/I481K/K483I, FcsCAE407R/I481R/K483T, and FcsFAE407R/I481K/K483R showed 9.96-, 10.58-, 4.25-, 6.49-, and 8.71-fold enhanced catalytic efficiency for degrading CoA thioesters of cinnamic acid, 4-methoxycinnamic acid, 4-hydroxycinnamic acid, caffeic acid, and ferulic acid, respectively. Molecular dynamics simulation illustrated that novel substrate-binding domains formed strong interaction forces with substrates’ methoxy/hydroxyl group and provided hydrophobic/alkaline catalytic surfaces. Five recombinant E. coli with FCS mutants were constructed with the maximum benzaldehyde, p-anisaldehyde, p-hydroxybenzaldehyde, protocatechualdehyde, and vanillin productivity of 6.2 ± 0.3, 5.1 ± 0.23, 4.1 ± 0.25, 7.1 ± 0.3, and 8.7 ± 0.2 mM/h, respectively. Hence, our study provided novel and efficient enzymes for the bioconversion of phenylpropanoid acids into aromatic aldehydes.

Simplification of Corticosteroids Biosynthetic Pathway by Engineering P450BM3

“…15 The B-factor and free energy analysis using MD simulation allows rapid identification of residues that obstruct substrate entry into the protein pocket as well as investigation of the forces exerted by residues within the pocket on the substrate. 34,39,40 Introduction of mutations R47I and R47L in P450BM3 can increase hydrophobicity and expand the channel, thereby facilitating better access of m-alkylphenol to the heme center. 28 Consistent with the findings of this study, P25A-P329A-E435D of VD-BM3 and A328G-L437G-L439G of FA11a1-BM3 can create larger, more accommodating channel surfaces that facilitate the access of more steroids to the heme center.…”

Efficient De Novo Biosynthesis of Curcumin in Escherichia coli by Optimizing Pathway Modules and Increasing the Malonyl-CoA Supply

“…In situ biosynthesis of methionine by strengthening the S-adenosyl methionine cycle could be utilized to reduce the cost of methionine and potentially improve the curcumin titer. − Modifying curcumin synthase to change its substrate specificity would be challenging because of the similarity of molecular structures of curcumin derivatives . As a potential alternative, feruloyl-CoA synthetase (FCS) has been modified to improve its catalytic efficiency toward phenylpropanoid acids by constructing substrate-binding domains; the modified FCS has a stronger preference for ferulic acid . In addition, curcumin could also be used to stress engineering strains and to explore curcumin transporters by transcriptomic analysis.…”

“…18 As a potential alternative, feruloyl-CoA synthetase (FCS) has been modified to improve its catalytic efficiency toward phenylpropanoid acids by constructing substrate-binding domains; the modified FCS has a stronger preference for ferulic acid. 49 In addition, curcumin could also be used to stress engineering strains and to explore curcumin transporters by transcriptomic analysis.…”

Efficient Production of Chlorogenic Acid in Escherichia coli Via Modular Pathway and Cofactor Engineering

“…Docking of substrates and enzymes was carried out using AutoDock 4.2.6. For the method of molecular docking, refer to the previous study . Visualization of proteins and molecules was done using PyMol 2.6.0.…”

“…For the method of molecular docking, refer to the previous study. 28 Visualization of proteins and molecules was done using PyMol 2.6.0. 2.6.…”