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

• Published in: Scientific reports 2018-04, Vol.8 (1), p.5856-11, Article 5856
• Main Authors: Kloss, Ramona, Limberg, Michael H., Mackfeld, Ursula, Hahn, Doris, Grünberger, Alexander, Jäger, Vera D., Krauss, Ulrich, Oldiges, Marco, Pohl, Martina
• Format: Article
• Language: English
• Subjects: 631/1647; 631/45/603; 631/61/252; Batch Cell Culture Techniques - methods; Biocatalysis; Biocatalysts; Bioreactors - microbiology; Cadaverine; Cadaverine - biosynthesis; Carboxy-Lyases - genetics; Carboxy-Lyases - isolation & purification; Carboxy-Lyases - metabolism; Cell culture; Corynebacterium glutamicum - metabolism; E coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli Proteins - genetics; Escherichia coli Proteins - isolation & purification; Escherichia coli Proteins - metabolism; Humanities and Social Sciences; Inclusion bodies; Inclusion Bodies - enzymology; Lysine; Lysine - metabolism; Lysine decarboxylase; Metabolic Engineering - methods; multidisciplinary; Purification; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-24070-2

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 ( Ec LDCc-CatIBs) to process L-lysine-containing culture supernatants from Corynebacterium glutamicum . Ec LDCc-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 Ec LDCc-CatIBs was demonstrated in (repetitive) batch experiments starting from L-lysine concentrations of 0.1 M and 1 M. Ec LDC-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 Ec LDCc-CatIBs, respectively. This resulted in DAP titres of up to 88.4 g l −1 and space-time yields of up to 660 g DAP l −1 d −1 per gram dry Ec LDCc-CatIBs. The new process for DAP production can therefore compete with the currently best fermentative process as described in the literature.