title: Engineering of Corynebacterium glutamicum for growth and L-lysine and lycopene production from N-acetyl-glucosamine.
abstract: Sustainable supply of feedstock has become a key issue in process development in microbial biotechnology. The workhorse of industrial amino acid production Corynebacterium glutamicum has been engineered towards utilization of alternative carbon sources. Utilization of the chitin-derived aminosugar N-acetyl-glucosamine (GlcNAc) for both cultivation and production with C. glutamicum has hitherto not been investigated. Albeit this organism harbors the enzymes N-acetylglucosamine-6-phosphatedeacetylase and glucosamine-6P deaminase of GlcNAc metabolism (encoded by nagA and nagB, respectively) growth of C. glutamicum with GlcNAc as substrate was not observed. This was attributed to the lack of a functional system for GlcNAc uptake. Of the 17 type strains of the genus Corynebacterium tested here for their ability to grow with GlcNAc, only Corynebacterium glycinophilum DSM45794 was able to utilize this substrate. Complementation studies with a GlcNAc-uptake deficient Escherichia coli strain revealed that C. glycinophilum possesses a nagE-encoded EII permease for GlcNAc uptake. Heterologous expression of the C. glycinophilum nagE in C. glutamicum indeed enabled uptake of GlcNAc. For efficient GlcNac utilization in C. glutamicum, improved expression of nagE with concurrent overexpression of the endogenous nagA and nagB genes was found to be necessary. Based on this strategy, C. glutamicum strains for the efficient production of the amino acid L-lysine as well as the carotenoid lycopene from GlcNAc as sole substrate were constructed.
authors.affiliation: Faculty of Biology and CeBiTec, Bielefeld University, 33501, Bielefeld, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Matano
authors.forename: Andreas
authors.initials: A
authors.lastname: Uhde
authors.forename: Jung-Won
authors.initials: JW
authors.lastname: Youn
authors.forename: Tomoya
authors.initials: T
authors.lastname: Maeda
authors.forename: Lina
authors.initials: L
authors.lastname: Clermont
authors.forename: Kay
authors.initials: K
authors.lastname: Marin
authors.forename: Reinhard
authors.initials: R
authors.lastname: Krämer
authors.forename: Volker F
authors.initials: VF
authors.lastname: Wendisch
authors.forename: Gerd M
authors.initials: GM
authors.lastname: Seibold
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D019747:Aldose-Ketose Isomerases / Q000235:genetics / Q000378:metabolism; D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D002338:Carotenoids / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005816:Genetic Complementation Test; D000077276:Lycopene; D008239:Lysine / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering
chemical_list: D026901:Membrane Transport Proteins; D002338:Carotenoids; D000581:Amidohydrolases; C018717:N-acetylglucosamine-6-phosphate deacetylase; C020582:glucosamine-6-phosphate isomerase; D019747:Aldose-Ketose Isomerases; D008239:Lysine; D000077276:Lycopene; D000117:Acetylglucosamine