title: Inactivation of phosphomannose isomerase gene abolishes sporulation and antibiotic production in Streptomyces coelicolor.
abstract: Phosphomannose isomerases (PMIs) in bacteria and fungi catalyze the reversible conversion of D-fructose-6-phosphate to D-mannose-6-phosphate during biosynthesis of GDP-mannose, which is the main intermediate in the mannosylation of important cell wall components, glycoproteins, and certain glycolipids. In the present study, the kinetic parameters of PMI from Streptomyces coelicolor were obtained, and its function on antibiotic production and sporulation was studied. manA (SCO3025) encoding PMI in S. coelicolor was deleted by insertional inactivation. Its mutant (S. coelicolor∆manA) was found to exhibit a bld-like phenotype. Additionally, S. coelicolor∆manA failed to produce the antibiotics actinorhodin and red tripyrolle undecylprodigiosin in liquid media. To identify the function of manA, the gene was cloned and expressed in Escherichia coli BL21 (DE3). The purified recombinant ManA exhibited PMI activity (K(cat)/K(m) (mM(-1) s(-1) = 0.41 for D-mannose-6-phosphate), but failed to show GDP-D-mannose pyrophosphorylase [GMP (ManC)] activity. Complementation analysis with manA from S. coelicolor or E. coli resulted in the recovery of bld-like phenotype of S. coelicolor∆manA. SCO3026, another ORF that encodes a protein with sequence similarity towards bifunctional PMI and GMP, was also tested for its ability to function as an alternate ManA. However, the purified protein of SCO3026 failed to exhibit both PMI and GMP activity. The present study shows that enzymes involved in carbohydrate metabolism could control cellular differentiation as well as the production of secondary metabolites.
authors.affiliation: Department of Microbial Engineering, College of Engineering, Konkuk University, Seoul, South Korea.
authors.forename: Thangamani
authors.initials: T
authors.lastname: Rajesh
authors.forename: Eunjung
authors.initials: E
authors.lastname: Song
authors.forename: Ji-Nu
authors.initials: JN
authors.lastname: Kim
authors.forename: Bo-Rahm
authors.initials: BR
authors.lastname: Lee
authors.forename: Eun-Jung
authors.initials: EJ
authors.lastname: Kim
authors.forename: Sung-Hee
authors.initials: SH
authors.lastname: Park
authors.forename: Yun-Gon
authors.initials: YG
authors.lastname: Kim
authors.forename: Dongwon
authors.initials: D
authors.lastname: Yoo
authors.forename: Hyung-Yeon
authors.initials: HY
authors.lastname: Park
authors.forename: Yun-Hui
authors.initials: YH
authors.lastname: Choi
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
authors.forename: Yung-Hun
authors.initials: YH
authors.lastname: Yang
mesh_terms: D000880:Anthraquinones / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D015870:Gene Expression; D005816:Genetic Complementation Test; D007700:Kinetics; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics* / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D011353:Prodigiosin / Q000031:analogs & derivatives / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013171:Spores, Bacterial / Q000254:growth & development*; D048372:Streptomyces coelicolor / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D000095622:Benzoisochromanequinones
chemical_list: D000880:Anthraquinones; D000900:Anti-Bacterial Agents; D008359:Mannose-6-Phosphate Isomerase; D011353:Prodigiosin; D011994:Recombinant Proteins; C047347:undecylprodigiosin; C013390:actinorhodin; D000095622:Benzoisochromanequinones