Engineering biotin prototrophic Corynebacterium glutamicum strains for amino acid, diamine and carotenoid production.
The Gram-positive Corynebacterium glutamicum is auxotrophic for biotin. Besides the biotin uptake system BioYMN and the transcriptional regulator BioQ, this bacterium possesses functional enzymes for the last three reactions of biotin synthesis starting from pimeloyl-CoA. Heterologous expression of bioF from the Gram-negative Escherichia coli enabled biotin synthesis from pimelic acid added to the medium, but expression of bioF together with bioC and bioH from E. coli did not entail biotin prototrophy. Heterologous expression of bioWAFDBI from Bacillus subtilis encoding another biotin synthesis pathway in C. glutamicum allowed for growth in biotin-depleted media. Stable growth of the recombinant was observed without biotin addition for eight transfers to biotin-depleted medium while the empty vector control stopped growth after the first transfer. Expression of bioWAFDBI from B. subtilis in C. glutamicum strains overproducing the amino acids l-lysine and l-arginine, the diamine putrescine, and the carotenoid lycopene, respectively, enabled formation of these products under biotin-depleted conditions. Thus, biotin-prototrophic growth and production by recombinant C. glutamicum were achieved.
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany. Electronic address: petra.peters-wendisch@uni-bielefeld.de.
P
P
Peters-Wendisch
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany.
S
S
Götker
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany.
S A E
SA
Heider
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany.
G
G
Komati Reddy
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany.
A Q
AQ
Nguyen
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany.
K C
KC
Stansen
Genetics of Prokaryotes, Faculty of Biology and CeBiTec, Bielefeld University, Bielefeld, Germany.
V F
VF
Wendisch
D000596:Amino Acids / Q000378:metabolism; D001710:Biotin / Q000235:genetics* / Q000378:metabolism*; D002338:Carotenoids / Q000378:metabolism; D048230:Corynebacterium glutamicum / Q000235:genetics* / Q000378:metabolism*; D003959:Diamines / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D000077276:Lycopene
D000596:Amino Acids; D003959:Diamines; D002338:Carotenoids; D001710:Biotin; D000077276:Lycopene
Amino acid production; Biotin biosynthesis; Carotenoid production; Corynebacterium glutamicum; Diamine production