Catalytically active inclusion bodies of L-lysine decarboxylase from E. coli for 1,5-diaminopentane production

Sci Rep. 2018 Apr 11;8(1):5856. doi: 10.1038/s41598-018-24070-2.

Abstract

Sustainable and eco-efficient alternatives for the production of platform chemicals, fuels and chemical building blocks require the development of stable, reusable and recyclable biocatalysts. Here we present a novel concept for the biocatalytic production of 1,5-diaminopentane (DAP, trivial name: cadaverine) using catalytically active inclusion bodies (CatIBs) of the constitutive L-lysine decarboxylase from E. coli (EcLDCc-CatIBs) to process L-lysine-containing culture supernatants from Corynebacterium glutamicum. EcLDCc-CatIBs can easily be produced in E. coli followed by a simple purification protocol yielding up to 43% dry CatIBs per dry cell weight. The stability and recyclability of EcLDCc-CatIBs was demonstrated in (repetitive) batch experiments starting from L-lysine concentrations of 0.1 M and 1 M. EcLDC-CatIBs exhibited great stability under reaction conditions with an estimated half-life of about 54 h. High conversions to DAP of 87-100% were obtained in 30-60 ml batch reactions using approx. 180-300 mg EcLDCc-CatIBs, respectively. This resulted in DAP titres of up to 88.4 g l-1 and space-time yields of up to 660 gDAP l-1 d-1 per gram dry EcLDCc-CatIBs. The new process for DAP production can therefore compete with the currently best fermentative process as described in the literature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Batch Cell Culture Techniques / methods
  • Biocatalysis
  • Bioreactors / microbiology
  • Cadaverine / biosynthesis*
  • Carboxy-Lyases / genetics
  • Carboxy-Lyases / isolation & purification
  • Carboxy-Lyases / metabolism*
  • Corynebacterium glutamicum / metabolism
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / isolation & purification
  • Escherichia coli Proteins / metabolism*
  • Inclusion Bodies / enzymology*
  • Lysine / metabolism
  • Metabolic Engineering / methods
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Escherichia coli Proteins
  • Recombinant Proteins
  • Carboxy-Lyases
  • lysine decarboxylase
  • Lysine
  • Cadaverine