Antibiotic overproduction in Streptomyces coelicolor A3 2 mediated by phosphofructokinase deletion.
Streptomycetes are exploited for production of a wide range of secondary metabolites, and there is much interest in enhancing the level of production of these metabolites. Secondary metabolites are synthesized in dedicated biosynthetic routes, but precursors and co-factors are derived from the primary metabolism. High level production of antibiotics in streptomycetes therefore requires engineering of the primary metabolism. Here we demonstrate this by targeting a key enzyme in glycolysis, phosphofructokinase, leading to improved antibiotic production in Streptomyces coelicolor A3(2). Deletion of pfkA2 (SCO5426), one of three annotated pfkA homologues in S. coelicolor A3(2), resulted in a higher production of the pigmented antibiotics actinorhodin and undecylprodigiosin. The pfkA2 deletion strain had an increased carbon flux through the pentose phosphate pathway, as measured by (13)C metabolic flux analysis, establishing the ATP-dependent PfkA2 as a key player in determining the carbon flux distribution. The increased pentose phosphate pathway flux appeared largely because of accumulation of glucose 6-phosphate and fructose 6-phosphate, as experimentally observed in the mutant strain. Through genome-scale metabolic model simulations, we predicted that decreased phosphofructokinase activity leads to an increase in pentose phosphate pathway flux and in flux to pigmented antibiotics and pyruvate. Integrated analysis of gene expression data using a genome-scale metabolic model further revealed transcriptional changes in genes encoding redox co-factor-dependent enzymes as well as those encoding pentose phosphate pathway enzymes and enzymes involved in storage carbohydrate biosynthesis.
Borodina
Irina
I
Center for Microbial Biotechnology, BioCentrum-DTU, Technical University of Denmark, 2800 Kongens Lyngby, Denmark.
Siebring
Jeroen
J
Department of Microbiology, Groningen Biotechnology and Biomolecular Sciences Institute, University of Groningen, P. O. Box 14, 9750 AA, Haren, The Netherlands.
Zhang
Jie
J
Center for Microbial Biotechnology, BioCentrum-DTU, Technical University of Denmark, 2800 Kongens Lyngby, Denmark.
Smith
Colin P
CP
Functional Genomics Laboratory, Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey GU2 7XH, United Kingdom.
van Keulen
Geertje
G
Biological Sciences, School of the Environment and Society, Swansea University, Swansea SA2 8PP, United Kingdom.
Dijkhuizen
Lubbert
L
Department of Microbiology, Groningen Biotechnology and Biomolecular Sciences Institute, University of Groningen, P. O. Box 14, 9750 AA, Haren, The Netherlands.
Nielsen
Jens
J
Center for Microbial Biotechnology, BioCentrum-DTU, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. Electronic address: jn@biocentrum.dtu.dk.
D000595:Amino Acid Sequence; D000880:Anthraquinones / Q000737:chemistry; D001483:Base Sequence; D002244:Carbon / Q000737:chemistry; D005636:Fructosephosphates / Q000737:chemistry; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D016678:Genome; D019298:Glucose-6-Phosphate / Q000737:chemistry; D008954:Models, Biological; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010732:Phosphofructokinase-1 / Q000235:genetics* / Q000502:physiology*; D011353:Prodigiosin / Q000031:analogs & derivatives / Q000737:chemistry; D048372:Streptomyces coelicolor / Q000378:metabolism*; D000095622:Benzoisochromanequinones
D000880:Anthraquinones; D002244:Carbon; D005636:Fructosephosphates; D019298:Glucose-6-Phosphate; D010732:Phosphofructokinase-1; D011353:Prodigiosin; C047347:undecylprodigiosin; C027618:fructose-6-phosphate; C013390:actinorhodin; D000095622:Benzoisochromanequinones