Systems metabolic engineering of xylose-utilizing Corynebacterium glutamicum for production of 1,5-diaminopentane

• Published in: Biotechnology journal Vol. 8; no. 5; pp. 557 - 570
• Main Authors: Buschke, Nele, Becker, Judith, Schäfer, Rudolf, Kiefer, Patrick, Biedendieck, Rebekka, Wittmann, Christoph
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01-05-2013; WILEY‐VCH Verlag
• Subjects: 13C metabolic flux analysis; Bacterial Proteins; Bio-polyamide; Cadaverine - analysis; Cadaverine - metabolism; Carbon Isotopes - metabolism; Corynebacterium glutamicum; Corynebacterium glutamicum - genetics; Corynebacterium glutamicum - metabolism; Elementary flux modes; Gene Regulatory Networks; Glucose - metabolism; Industrial Microbiology; Membrane Transport Proteins; Metabolic Engineering - methods; Metabolic Networks and Pathways; Systems Biology - methods; Transcriptome; Trehalose - metabolism; Xylose; Xylose - metabolism
• ISSN: 1860-6768; 1860-7314
• DOI: 10.1002/biot.201200367

The sustainable production of industrial platform chemicals is one of the great challenges facing the biotechnology field. Ideally, fermentation feedstocks would rather rely on industrial waste streams than on food‐based raw materials. Corynebacterium glutamicum was metabolically engineered to produce the bio‐nylon precursor 1,5‐diaminopentane from the hemicellulose sugar xylose. Comparison of a basic diaminopentane producer strain on xylose and glucose feedstocks revealed a 30% reduction in diaminopentane yield and productivity on the pentose sugar. The integration of in vivo and in silico metabolic flux analysis by 13C and elementary modes identified bottlenecks in the pentose phosphate pathway and the tricarboxylic acid cycle that limited performance on xylose. By the integration of global transcriptome profiling, this could be specifically targeted to the tkt operon, genes that encode for fructose bisphosphatase (fbp) and isocitrate dehydrogenase (icd), and to genes involved in formation of lysine (lysE) and N‐acetyl diaminopentane (act). This was used to create the C. glutamicum strain DAP‐Xyl1 icdGTG Peftufbp Psodtkt Δact ΔlysE. The novel producer, designated DAP‐Xyl2, exhibited a 54% increase in product yield to 233 mmol mol–1 and a 100% increase in productivity to 1 mmol g–1 h–1 on the xylose substrate. In a fed‐batch process, the strain achieved 103 g L–1 of diaminopentane from xylose with a product yield of 32%. Xylose utilization is currently one of the most relevant metabolic engineering subjects. In this regard, the current work is a milestone in industrial strain engineering of C. glutamicum.
See accompanying commentary by Hiroshi Shimizu DOI: 10.1002/biot.201300097
The sustainable production of chemicals from renewable, non‐food raw materials is one of the great challenges in industrial biotechnology. In this article, the authors report reprogramming of Corynebacterium glutamicum for efficient 1,5‐diaminopentane production from xylose via systems metabolic engineering. Reaching a yield of 32% and a maximum titer of 103 gL–1, the current work reaches a milestone in industrial strain engineering with C. glutamicum.