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.

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BackgroundRecent interest has been focused on the production of platform chemicals from renewable biomass due to increasing concerns on global warming and depletion of fossil fuel reserves. Microbial production of platform chemicals in biorefineries has been suggested to be a promising solution for these problems. Gamma-aminobutyrate (GABA), a versatile bulk chemical used in food and pharmaceutical industry, is also used as a key monomer for nylon 4. GABA can be biologically produced by decarboxylation of glutamate.ResultsIn this study, we examined high glutamate-producing Corynebacterium glutamicum strains as hosts for enhanced production of GABA from glucose and xylose as carbon sources. An Escherichia coli gadB mutant with a broad pH range of activity and E. coli xylAB genes were expressed under the control of a synthetic H36 promoter. When empty fruit bunch (EFB) solution was used as carbon source (45 g/L glucose and 5 g/L xylose), 12.54 ± 0.07 g/L GABA was produced by recombinant C. glutamicum H36GD1852 expressing E. coli gadB mutant gene and xylAB genes. Batch fermentation of the same strain resulted in the production of 35.47 g/L of GABA when EFB solution was added to support 90 g/L glucose and 10 g/L xylose.ConclusionsThis is the first report of GABA production by recombinant C. glutamicum strains from co-utilization of glucose and xylose from EFB solution. Recombinant C. glutamicum strains developed in this study should be useful for an efficient and sustainable production of GABA from lignocellulosic biomasses.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0977-9) contains supplementary material, which is available to authorized users.

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Here, we report on a highly sensitive method for the detection of P(3HB) accumulation in Escherichia coli cells based on the automated flow cytometry system using fluorescent dyes. E. coli containing P(3HB) were stained with either BODIPY or Nile red fluorescent dye, and their staining properties were analyzed under a variety of conditions. Compared with Nile red, BODIPY was much more sensitive in staining P(3HB) and overall demonstrated a more rapid staining of cells, a greater resistance to photobleaching, and greater cell viability. In addition, we also successfully monitored heterogeneity in P(3HB) accumulation within a cell population using BODIPY staining and flow cytometry. We believe this optimized staining method using BODIPY in combination with screening by high-speed flow cytometer will be helpful in the engineering of host cells toward an enhanced production of bioplastics.