A High-Efficiency Artificial Synthetic Pathway for 5-Aminovalerate Production From Biobased L-Lysine in Escherichia coli

Abstract

Bioproduction of 5-aminovalerate (5AVA) from renewable feedstock can support a sustainable biorefinery process to produce bioplastics, such as nylon 5 and nylon 56. In order to achieve the biobased production of 5AVA, a 2-keto-6-aminocaproate-mediated synthetic pathway was established. Combination of L-Lysine α-oxidase from Scomber japonicus, α-ketoacid decarboxylase from Lactococcus lactis and aldehyde dehydrogenase from Escherichia coli could achieve the biosynthesis of 5AVA from biobased L-Lysine in E. coli. The H2O2 produced by L-Lysine α-oxidase was decomposed by the expression of catalase KatE. Finally, 52.24 g/L of 5AVA were obtained through fed-batch biotransformation. Moreover, homology modeling, molecular docking and molecular dynamic simulation analyses were used to identify mutation sites and propose a possible trait-improvement strategy: the expanded catalytic channel of mutant and more hydrogen bonds formed might be beneficial for the substrates stretch. In summary, we have developed a promising artificial pathway for efficient 5AVA synthesis.

Evaluation of Heterologous Biosynthetic Pathways for Methanol-Based 5-Aminovalerate Production by Thermophilic Bacillus methanolicus

“…Therefore, the precursor levels in the B. methanolicus strains should not be a limiting factor for production of 5AVA. The RaiP-mediated production is mainly utilized in the L-lysine bioconversion approach, utilizing E. coli strains as whole-cell biocatalysts (Cheng et al, 2018(Cheng et al, , 2020(Cheng et al, , 2021 where high concentrations of the precursor were used; for example, the molar yield of 0.942 was obtained from 120 g l −1 Llysine (Park et al, 2014). However, construction and testing of the B. methanolicus strains M168-20_RaiP Sj , M168-20_RaiP Ps , and M168-20_RaiP Tv (Table 4), based on the L-lysine-over producing mutant M168-20 (Brautaset et al, 2010), did not result in any improved 5AVA production (data not shown).…”

“…Recently, an artificial synthetic pathway for the biosynthesis of 5AVA in E. coli was developed, consisting of three steps: conversion of L -lysine to α-ketolysine via RaiP, decarboxylation of α-ketolysine to produce 5-aminopentanal via α-ketoacid decarboxylase, and oxidation of 5-aminopentanal to 5AVA via aldehyde dehydrogenase. The expression of the artificial pathway resulted in a yield increase of 774% compared to the single gene pathway ( Cheng et al, 2021 ). This approach is potentially a feasible strategy we have shown in our study that E. coli -derived PatD is active as a 5-aminopentanal dehydrogenase in B. methanolicus and participates in 5AVA biosynthesis (see Section “The PatA Pathway Supports 5AVA Accumulation in B. methanolicus ).…”

Utilization of a Wheat Sidestream for 5-Aminovalerate Production in Corynebacterium glutamicum

“…In the first pathway, l -lysine is converted to 5AVA by l -lysine-α-oxidase (RaiP) from Scomber japonicas by oxidative deamination and a spontaneous decarboxylation step (Cheng et al, 2020). Recently, an alternative synthetic route was established starting with RaiP, but the intermediate 2-keto-6-aminocaproate is converted by α-ketoacid decarboxylase (KivD) from Lactococcus lactis and aldehyde dehydrogenase (PadA) from Escherichia coli to 5AVA ( Cheng et al, 2021 ). The second pathway to 5AVA combines oxidative decarboxylation by l -lysine monooxygenase (DavA) using molecular oxygen followed by desamidation by γ-aminovaleramidase (DavB) from P. putida ( Adkins et al, 2013 ).…”

Coproduction of 5-Aminovalerate and δ-Valerolactam for the Synthesis of Nylon 5 From L-Lysine in Escherichia coli

“…The monomers of polyamides are primarily dicarboxylic acids, diamines, lactams, and ω-amino acids ( Radzik et al, 2019 ). Examples of these main platform chemicals range from succinate ( Zhang et al, 2009 ), glutarate ( Zhao et al, 2018 ), to adipate ( Wang F. et al, 2020 ) for dicarboxylic acids; from putrescine, cadaverine ( Rui et al, 2020 ; Xue et al, 2020 ), to 1,6-hexanediamine for diamines; from δ-valerolactam ( Zhang et al, 2017a ), to ε-caprolactam ( Thompson et al, 2020 ) for lactams; from 3-hydroxybutyrate ( Atakav et al, 2021 ; Mierziak et al, 2021 ; Schmid et al, 2021 ), 2-hydroxybutyrate ( Tian et al, 2021 ), to 3-hydroxyhexanoate ( Harada et al, 2021 ) for hydroxyl acids; and from 4-aminobutyrate, 5AVA ( Cheng et al, 2021b ), to 6-aminocaproate ( Turk et al, 2016 ) for ω-amino acids. In this respect, also 5AVA ( Adkins et al, 2013 ) and δ-valerolactam ( Xu et al, 2020 ) are attractive C5 platform chemicals for the production of biopolyamides from renewable biomass.…”

“…Interestingly, Cheng et al proposed that the titer of 5AVA could be improved to 29.12 g/L by adding 4% (v/v) ethanol and 10 mM H 2 O 2 ( Cheng et al, 2018b ). Independently, a three-step route based on RaiP, α-ketoacid decarboxylase (KivD) from Lactococcus lactis , and aldehyde dehydrogenase (PadA) from Escherichia coli ( E. coli ) was established in E. coli with 5AVA titer up to about 52.24 g/L ( Cheng et al, 2021b ).…”