Enzymatic Synthesis of ω-Hydroxydodecanoic Acid By Employing a Cytochrome P450 from Limnobacter sp. 105 MED

Abstract

ω-Hydroxylated fatty acids are valuable and versatile building blocks for the production of various adhesives, lubricants, cosmetic intermediates, etc. The biosynthesis of ω-hydroxydodecanoic acid from vegetable oils is one of the important green pathways for their chemical-based synthesis. In the present study, the novel monooxygenase CYP153AL.m from Limnobacter sp. 105 MED was used for the whole-cell biotransformations. We constructed three-component system that was comprised of CYP153AL.m, putidaredoxin and putidaredoxin reductase from Pseudomonas putida. This in vivo study demonstrated that CYP153AL.m is a powerful catalyst for the biosynthesis of ω-hydroxydodecanoic acid. Under optimized conditions, the application of a solid-state powdered substrate rather than a substrate dissolved in DMSO significantly enhanced the overall reaction titer of the process. By employing this efficient system, 2 g/L of 12-hydroxydodecanoic acid (12-OHDDA) was produced from 4 g/L of its corresponding fatty acid, which was namely dodecanoic acid. Furthermore, the system was extended to produce 3.28 g/L of 12-OHDDA using 4 g/L of substrate by introducing native redox partners. These results demonstrate the utility of CYP153AL.m-catalyzed biotransformations in the industrial production of 12-OHDDA and other valuable building blocks.

Multi-enzymatic cascade reactions with Escherichia coli-based modules for synthesizing various bioplastic monomers from fatty acid methyl esters

“…11 However, saturated fatty acids can be converted directly into ω-HFAs by monooxygenases such as P450 and AlkBGT without the loss of carbon atoms, while unsaturated fatty acids require 4 enzymes ( vide supra ). 16,17 Also the broad substrate specificity of monooxygenases enables the biosynthesis of various carbon chain length products. 18 However, the production of various biopolymer monomers from saturated fatty acids has not been extensively studied, due to the difficulty of functionalization of non-activated carbon.…”

Biosynthesis of Medium-Chain ω-Hydroxy Fatty Acids by AlkBGT of Pseudomonas putida GPo1 With Native FadL in Engineered Escherichia coli

“…For example, these include CYP52A13 and CYP52A17 (Craft et al, 2003; Eschenfeldt et al, 2003), CYP4 (Fer et al, 2008), CYP704B2 (Li et al, 2010), CYP102A1 (Xiao et al, 2018), CYP153 (Bordeaux et al, 2012), and the CYP153A M.aq. -CPR BM3 fusion protein (Scheps et al, 2013; Ahsan et al, 2018; Joo et al, 2019). Some of these enzymes can ω-oxidize terminal methyl groups and have been applied to ω-HFA production, but there is a limitation, in that most of these enzymes are specific for chain lengths of ≥12 (Picataggio et al, 1992).…”

Recent Advances on Biocatalysis and Metabolic Engineering for Biomanufacturing

“…Joo et al reported the biosynthesis of ω-hydroxydodecanoic acid via whole-cell biotransformations using a novel monooxygenase CYP153AL.m from Limnobacter sp. 105 MED [2]. ω-Aminododecanoic acid can be used as Nylon 12 monomers.…”