Efficient production of phenylpropionic acids by an amino‐group‐transformation biocatalytic cascade

Abstract

Phenylpropionic acids are commonly used in the synthesis of pharmaceuticals, cosmetics, and fine chemicals. However, the synthesis of phenylpropionic acids faces the challenges of high cost of substrates and a limited range of products. Here, we present an artificially designed amino-group-transformation biocatalytic process, which uses simple phenols, pyruvate, and ammonia to synthesize diverse phenylpropionic acids. This biocatalytic cascade comprises an amino-group-introduction module and three amino-group-transformation modules, and operates in a modular assembly manner. Escherichia coli catalysts coexpressing enzymes from different modules achieve whole-cell simultaneous one-pot transformations of phenols into the corresponding phenylpropionic acids including (S)-α-amino acids, α-keto acids, (R)-αamino acids, and (R)-β-amino acids. With cofactor recycling, protein engineering, and transformation optimization, four (S)-α-amino acids, four α-keto acids, four (R)-αamino acids, and four (R)-β-amino acids are synthesized with good conversion (68-99%) and high enantioselectivities (>98%). Therefore, the amino-grouptransformation concept provides a universal and efficient tool for synthesizing diverse products.

One-Pot Enzymatic–Chemical Cascade Route for Synthesizing Aromatic α-Hydroxy Ketones

“…A cascade reaction was then designed with PDC, carbonyl reductase (SCR or RCR), and glucose dehydrogenase (GDH) to produce optically pure ( S )- and ( R )-1-phenyl-1,2-ethanediol [( S )- or ( R )-PED] using 1a and 2 as substrates (Figure ). Additionally, based on the literature mining and the enzyme activity, the NADH-dependent SCR from Gluconobacter oxydans , the RCR from Bacillus subtilis 168, and Bm GDH from Bacillus megatherium were used to construct engineered E. coli strains harboring pET-Deut- Go SCR- Bm GDH or pET-Deut- Bs RCR- Bm GDH plasmids ( E. coli WL03 and E. coli WL04, respectively) (Figure S5 and Table ).…”

“…CtPDC1 WT 6.9 ± 0. mining and the enzyme activity, the NADH-dependent SCR from Gluconobacter oxydans, the RCR from Bacillus subtilis 168, 8 and BmGDH from Bacillus megatherium 20 were used to construct engineered E. coli strains harboring pET-Deut-GoSCR-BmGDH or pET-Deut-BsRCR-BmGDH plasmids (E. coli WL03 and E. coli WL04, respectively) (Figure S5 and Table 4).…”

Multienzymatic Cascade for Synthesis of Hydroxytyrosol via Two-Stage Biocatalysis

“…E. coli BL21 (DE3) cells harboring the desired genes were cultured at 37 °C in 100 mL of LB (Luria−Bertani) medium supplemented with the appropriate antibiotics (50 mg/L chloramphenicol, 50 mg/L streptomycin, 100 mg/L ampicillin, 50 mg/L kanamycin, or a combination thereof). 25 IPTG was added to the culture at a final concentration of 0.1 mM once the OD 600 nm (optical density at 600 nm) reached 0.4−0.6. The culture was maintained at 25 °C with constant shaking for an additional 16 h to induce the desired enzymes to overexpress.…”

A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine

“…In this process, DL-HPG is first esterified with thionyl chloride to generate DL-HPG methyl ester, followed by hydrolysis to generate D-HPG (Zhang et al 2011 ). Alternatively, enzyme catalysis provides a promising and efficient approach for synthesizing chiral chemicals (Wiltschi et al 2020 ; Wu et al 2021 ; Xue et al 2018 ), such as ( R )-β-tyrosine, ( R )-phenyllactic acid, and l -homophenylalanine, which are commonly used in the synthesis of pharmaceuticals, cosmetics, and fine chemicals (Song et al 2018 ; Wang et al 2020 ). Accordingly, an enzymatic catalysis strategy involving dual-enzyme synthesis was developed, with this process comprising ring opening of dl -hydroxyphenylhydantoin (DL-HPH) and hydrolysis, catalyzed by d -hydantoinase (Hase; EC 3.5.2.2) and N -carbamoyl- d -amino-acid hydrolase (Case; EC 3.5.1.77), respectively (Aranaz et al 2015 ; Diez et al 2015 ; Liu et al 2019 ).…”