Effects of methylmalonyl-CoA mutase gene knockouts on erythromycin production in carbohydrate-based and oil-based fermentations of Saccharopolyspora erythraea

  title={Effects of methylmalonyl-CoA mutase gene knockouts on erythromycin production in carbohydrate-based and oil-based fermentations of Saccharopolyspora erythraea},
  author={Andrew R. Reeves and Igor A. Brikun and William H. Cernota and Benjamin I. Leach and Melissa C. Gonzalez and J. Mark Weber},
  journal={Journal of Industrial Microbiology and Biotechnology},
In carbohydrate-based fermentations of Saccharopolyspora erythraea, a polar knockout of the methylmalonyl-CoA mutase (MCM) gene, mutB, improved erythromycin production an average of 126% (within the range of 102–153% for a 0.95 confidence interval). In oil-based fermentations, where erythromycin production by the wild-type strain averages 184% higher (141–236%, 0.95 CI) than in carbohydrate-based fermentations, the same polar knockout in mutB surprisingly reduced erythromycin production by 66… 
Engineering of the methylmalonyl-CoA metabolite node for increased erythromycin production in oil-based fermentations of Saccharopolyspora erythraea
The combined results showed that increased erythromycin production only occurred in strain FL2385 containing a duplication of the entire MCM operon including mutR and a predicted stem-loop structure overlapping the 3′ terminus of the mutR coding sequence.
An erythromycin process improvement using the diethyl methylmalonate-responsive (Dmr) phenotype of the Saccharopolyspora erythraea mutB strain
The Saccharopolyspora erythraea mutB knockout strain, FL2281, was found to carry a diethyl methylmalonate-responsive (Dmr) phenotype, which represents a new class of strain improvement phenotype and a theory to explain the biochemical mechanism for the Dmr phenotype is proposed.
Blocking the flow of propionate into TCA cycle through a mutB knockout leads to a significant increase of erythromycin production by an industrial strain of Saccharopolyspora erythraea
A high erystromycin producing mutant strain Saccharopolyspora erythraea HL3168 E3-ΔmutB was constructed by deleting mutB (SACE_5639) gene encoding the beta subunit of methylmalonyl-CoA mutase of an industrial strain of S. eryTHraeaHL3 168 E3 to characterize the physiological parameters.
Fermentation optimization and industrialization of recombinant Saccharopolyspora erythraea strains for improved erythromycin a production
Analysis of intra- and extracellular metabolites and key enzyme activities in central carbon metabolism revealed that the pool of TCA cycle intermediates was enhanced by the addition of corn steep liquor and induced an increase in erythromycin biosynthesis.
Effects of the Methylmalonyl-CoA Metabolic Pathway on Ansamitocin Production in Actinosynnema pretiosum
Investigating two key enzymes related to the methylmalonyl-CoA metabolic pathway suggested that eliminating the bypass pathways and favoring the precursor synthetic pathway could effectively increase ansamitocin production in A. pretiosum.
Knockout of the Erythromycin Biosynthetic Cluster Gene, eryBI, Blocks Isoflavone Glucoside Bioconversion during Erythromycin Fermentations in Aeromicrobium erythreum but Not in Saccharopolyspora erythraea
This study showed that isoflavone metabolism could be blocked in A. erythreum by ery BI knockout but that eryBI knockout was not sufficient to block is oflav one metabolism in S. eries, suggesting that other β-glucosidases are present.
PccD Regulates Branched-Chain Amino Acid Degradation and Exerts a Negative Effect on Erythromycin Production in Saccharopolyspora erythraea
The results demonstrated that PccD controlled the supply of precursors for biosynthesis of erythromycin via regulating the BCAA degradation and propionyl-CoA assimilation and exerted a negative effect on erystromycin production.
A combined approach of classical mutagenesis and rational metabolic engineering improves rapamycin biosynthesis and provides insights into methylmalonyl-CoA precursor supply pathway in Streptomyces hygroscopicus ATCC 29253
The results demonstrated that the combined approach involving traditional mutagenesis and metabolic engineering could be successfully applied to the diagnosis of yield-limiting factors and the enhanced production of industrially and clinically important polyketide compounds.


Crotonyl-coenzyme A reductase provides methylmalonyl-CoA precursors for monensin biosynthesis by Streptomyces cinnamonensis in an oil-based extended fermentation.
The results demonstrate that the relative contributions of different pathways and enzymes to providing polyketide precursors are thus dependent upon the fermentation conditions and the generally accepted pathways for providing methylmalonyl-CoA forpolyketide production may not be significant.
Insertional Inactivation of Methylmalonyl Coenzyme A (CoA) Mutase and Isobutyryl-CoA Mutase Genes in Streptomyces cinnamonensis: Influence on Polyketide Antibiotic Biosynthesis
C-labeling experiments show that in both mutants butyrate and acetoacetate may be incorporated into the propionate units in monensin A without cleavage to acetate units, suggesting that n-butyryl-CoA may be converted into methylmalonyl- coenzyme A through a carbon skeleton rearrangement for which neither ICM nor MCM alone is essential.
MeaA, a Putative Coenzyme B12-Dependent Mutase, Provides Methylmalonyl Coenzyme A for Monensin Biosynthesis in Streptomyces cinnamonensis
The results demonstrate that the meaA gene product is significantly involved in methylmalonyl-CoA production in S. cinnamonensis and that under the tested conditions the presence of both MeaA and ICM is crucial for monensin production in the WD2 strain.
Molecular analysis and heterologous expression of the gene encoding methylmalonyl—coenzyme a mutase from rifamycin SV-producing strain Amycolatopsis mediterranei U32
The structural gene for MCM from rifamycin SV—producing strain Amycolatopsis mediterranei U32 was isolated by using a heterologous gene probe encoding the MCM of Streptomyces cinnamonensis and a striking cross-species conservation of gene order suggested that ORF5 could also be involved in the metabolism of methylmalonyl-CoA.
MeaB Is a Component of the Methylmalonyl-CoA Mutase Complex Required for Protection of the Enzyme from Inactivation*
It is demonstrated that MeaB forms a complex with methylmalonyl-CoA mutase and stimulates in vitro mutase activity, which supports the hypothesis that MeAB functions to protect methylmalonic aciduria-related mutase from irreversible inactivation.
Role of Crotonyl Coenzyme A Reductase in Determining the Ratio of Polyketides Monensin A and Monensin B Produced byStreptomyces cinnamonensis
It is demonstrated that CCR plays a significant role in providing butyryl-CoA for monensin A biosynthesis and is present in wild-type S. cinnamonensis C730.1 at a level sufficient that the availability of the appropriate substrate (crotonyl- CoA) is limiting.
Ethyl-substituted erythromycin derivatives produced by directed metabolic engineering.
  • D. Stassi, S. Kakavas, L. Katz
  • Biology, Chemistry
    Proceedings of the National Academy of Sciences of the United States of America
  • 1998
A previously unknown chemical structure, 6-desmethyl-6-ethylerythromycin A (6-ethylErA), was produced through directed genetic manipulation of the erythromycin (Er)-producing organism
Engineering precursor flow for increased erythromycin production in Aeromicrobium erythreum.
Organization of a cluster of erythromycin genes in Saccharopolyspora erythraea.
The new Ery phenotype, EryH, was marked by the accumulation of the intermediate 6-deoxyerythronolide B (DEB), suggesting a defect in the operation of the C-6 hydroxylase system, and a block in the synthesis or addition reactions for the first sugar group.
Genetic and biochemical characterization of the α and β components of a propionyl-CoA carboxylase complex of Streptomyces coelicolor A3(2)
Heterologous expression of accA1, accA2 and pccB in Escherichia coli and in vitro reconstitution of enzyme activity confirmed that PccB is the β subunit of a propionyl-CoA carboxylase and that either AccA1 or AccA2 could act as the α component of this enzyme complex.