title: Production and glucosylation of C50 and C 40 carotenoids by metabolically engineered Corynebacterium glutamicum.
abstract: The yellow-pigmented soil bacterium Corynebacterium glutamicum ATCC13032 is accumulating the cyclic C50 carotenoid decaprenoxanthin and its glucosides. Carotenoid pathway engineering was previously shown to allow for efficient lycopene production. Here, engineering of C. glutamicum for production of endogenous decaprenoxanthin as well as of the heterologous C50 carotenoids C.p.450 and sarcinaxanthin is described. Plasmid-borne overexpression of genes for lycopene cyclization and hydroxylation from C. glutamicum, Dietzia sp., and Micrococcus luteus, in a lycopene-producing platform strain constructed here, resulted in accumulation of these three C50 carotenoids to concentrations of about 3-4 mg/g CDW. Chromosomal deletion of a putative carotenoid glycosyltransferase gene cg0730/crtX in these strains entailed production of non-glucosylated derivatives of decaprenoxanthin, C.p.450, and sarcinaxanthin, respectively. Upon introduction of glucosyltransferase genes from M. luteus, C. glutamicum, and Pantoea ananatis, these hydroxylated C50 carotenoids were glucosylated. We here also demonstrate production of the C40 carotenoids β-carotene and zeaxanthin in recombinant C. glutamicum strains and co-expression of the P. ananatis crtX gene was used to obtain glucosylated zeaxanthin. Together, our results show that C. glutamicum is a potentially valuable host for production of a wide range of glucosylated C40 and C50 carotenoids.
authors.affiliation: Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitätsstr. 25, 33615, Bielefeld, Germany.
authors.forename: Sabine A E
authors.initials: SA
authors.lastname: Heider
authors.forename: Petra
authors.initials: P
authors.lastname: Peters-Wendisch
authors.forename: Roman
authors.initials: R
authors.lastname: Netzer
authors.forename: Marit
authors.initials: M
authors.lastname: Stafnes
authors.forename: Trygve
authors.initials: T
authors.lastname: Brautaset
authors.forename: Volker F
authors.initials: VF
authors.lastname: Wendisch
mesh_terms: D000192:Actinomycetales / Q000201:enzymology / Q000235:genetics; D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D006031:Glycosylation; D060847:Metabolic Engineering / Q000379:methods*; D008837:Micrococcus / Q000201:enzymology / Q000235:genetics; D020636:Pantoea / Q000201:enzymology / Q000235:genetics; D024341:Xanthophylls / Q000378:metabolism*
chemical_list: D024341:Xanthophylls; C014802:sarcinaxanthin; C011431:decaprenoxanthin