Metabolic Engineering of Corynebacterium glutamicum for the High-Level Production of Cadaverine That Can Be Used for the Synthesis of Biopolyamide 510

Abstract

Fermentative production of cadaverine from renewable resources may support a sustainable biorefinery process to produce carbon-neutral nylons such as biopolyamide 510 (PA510). Cost-competitive production of cadaverine is a key factor in the successful commercialization of PA510. In this study, an integrated biological and chemical process involving cadaverine biosynthesis, purification, and its polymerization with sebacic acid was developed to produce bio-PA510. To stably express ldcC from Escherichia coli in an engineered Corynebacterium glutamicum PKC strain, an expired industrial L-lysine-producing strain, ldcC, was integrated into the chromosome of the C. glutamicum PKC strain by disrupting lysE and controlling its expression via a strong synthetic H30 promoter. Cadaverine was produced at a concentration of 103.78 g/L, the highest titer to date, from glucose by fed-batch culture of this engineered C. glutamgicum PKC strain. Fermentation-derived cadaverine was purified to polymer-grade biocadaverine with high purity (99%) by solvent extraction with chloroform and two-step distillation. Finally, biobased PA510 with good thermal properties (T m 215 °C and T c 158 °C) was produced by polymerization of purified cadaverine with sebacic acid. The hybrid biorefinery process combining biological and chemical processes demonstrated in this study is a useful platform for producing biobased chemicals and polymers.

Recent advances in metabolic engineering of Corynebacterium glutamicum as a potential platform microorganism for biorefinery

“…Because lysine decarboxylase is a key enzyme for cadaverine production, codon optimization, chromosomal integration ( bioD for ATP‐dependent dethiobiotin synthetase; lysE for l ‐lysine exporter), and expression under strong promoters (P tuf and P H30 ) greatly affect the final titer of cadaverine produced by recombinant C. glutamicum (Fig. ) . When ldcC from E. coli W3110 was codon‐optimized for C. glutamicum and integrated into the bioD locus for the chromosomal‐based expression of LDC, the final titer of cadaverine was 88 g L −1 from fed‐batch fermentation.…”

“…To improve the production of cadaverine, industrial l ‐lysine‐overproducing C. glutamicum PKC was used as a host for the chromosomal integration of E. coli ldcC under the control of a strong H30 promoter into lysE locus. This enabled the stable expression of LDC and deletion of l ‐lysine export activity, which led to increased cadaverine production (104 g L −1 ) . To improve the export of cadaverine, the overexpression of a putative putrescine exporter gene ( cgmA) enhanced the extracellular cadaverine concentration .…”

Development of an integrated process for the production of high‐purity cadaverine from lysine decarboxylase

“…21 Kim et al reported that ldcC from E. coli was integrated into the chromosome of the C. glutamicum PKC strain, which was expressed through a strong synthetic promoter H30, and the engineered strain produced a concentration of 103.78 g L -1 cadaverine from glucose by fed-batch culture. 22 To our understanding, these research results about the bioproduction of cadaverine suggest their suitability for scaling-up to industrial production.…”

Cadaverine Production From L-Lysine With Chitin-Binding Protein-Mediated Lysine Decarboxylase Immobilization

“…Polyamides (PA) are an essential polymer and widely used in engineering plastics, sportswear, sutures and catheters, owing to the excellent mechanical and thermal strength (Park et al, 2014;Jiang and Loos, 2016;Kim et al, 2018). Polyamides are mostly produced from chemicals extracted from fossil fuels (6.6 million tons per year), which contributes to the greenhouse effect and serious environmental pollution (Hong et al, 2004;Kind et al, 2011).…”