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ACS Synth Biol ; 5(1): 65-73, 2016 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-26511532

RESUMO

Nylon-6 is a bulk polymer used for many applications. It consists of the non-natural building block 6-aminocaproic acid, the linear form of caprolactam. Via a retro-synthetic approach, two synthetic pathways were identified for the fermentative production of 6-aminocaproic acid. Both pathways require yet unreported novel biocatalytic steps. We demonstrated proof of these bioconversions by in vitro enzyme assays with a set of selected candidate proteins expressed in Escherichia coli. One of the biosynthetic pathways starts with 2-oxoglutarate and contains bioconversions of the ketoacid elongation pathway known from methanogenic archaea. This pathway was selected for implementation in E. coli and yielded 6-aminocaproic acid at levels up to 160 mg/L in lab-scale batch fermentations. The total amount of 6-aminocaproic acid and related intermediates generated by this pathway exceeded 2 g/L in lab-scale fed-batch fermentations, indicating its potential for further optimization toward large-scale sustainable production of nylon-6.


Assuntos
Caprolactama/análogos & derivados , Engenharia Metabólica/métodos , Polímeros/síntese química , Adipatos/metabolismo , Ácido Aminocaproico/metabolismo , Técnicas de Cultura Celular por Lotes , Caprolactama/síntese química , Cromatografia Líquida , Escherichia coli/genética , Escherichia coli/metabolismo , Fermentação , Metaboloma , Ácidos Pimélicos/metabolismo , Proteômica , Espectrometria de Massas em Tandem , Ácidos Tricarboxílicos/metabolismo
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