Using Chemobiosynthesis and Synthetic Mini-Polyketide Synthases To Produce Pharmaceutical Intermediates in Escherichia coli

Kosan Biosciences Inc., Hayward, California 94545, USA.
Applied and Environmental Microbiology (Impact Factor: 3.67). 08/2010; 76(15):5221-7. DOI: 10.1128/AEM.02961-09
Source: PubMed


Recombinant microbial whole-cell biocatalysis is a valuable approach for producing enantiomerically pure intermediates for
the synthesis of complex molecules. Here, we describe a method to produce polyketide intermediates possessing multiple stereogenic
centers by combining chemobiosynthesis and engineered mini-polyketide synthases (PKSs). Chemobiosynthesis allows the introduction
of unnatural moieties, while a library of synthetic bimodular PKSs expressed from codon-optimized genes permits the introduction
of a variety of ketide units. To validate the approach, intermediates for the synthesis of trans-9,10-dehydroepothilone D were generated. The designer molecules obtained have the potential to greatly reduce the manufacturing
cost of epothilone analogues, thus facilitating their commercial development as therapeutic agents.

Download full-text


Available from: John R Carney, Jan 23, 2014
  • Source
    • "Next, more than three million bases of PKS genes were tested to validate the platform. These efforts produced a variety of novel valuable compounds (Menzella et al., 2007; Menzella et al., 2010). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The efficient production of functional proteins in heterologous hosts is one of the major bases of modern biotechnology. Unfortunately, many genes are difficult to express outside their original context. Due to their apparent "silent" nature, synonymous codon substitutions have long been thought to be trivial. In recent years, this dogma has been refuted by evidence that codon replacement can have a significant impact on gene expression levels and protein folding. In the past decade, considerable advances in the speed and cost of gene synthesis have facilitated the complete redesign of entire gene sequences, dramatically improving the likelihood of high protein expression. This technology significantly impacts the economic feasibility of microbial-based biotechnological processes by, for example, increasing the volumetric productivities of recombinant proteins or facilitating the redesign of novel biosynthetic routes for the production of metabolites. This review discusses the current applications of this technology, particularly those regarding the production of small molecules and industrially relevant recombinant enzymes. Suggestions for future research and potential uses are provided as well.
    Full-text · Article · Feb 2014 · Frontiers in Microbiology
  • [Show abstract] [Hide abstract]
    ABSTRACT: Antibiotics and mycotoxins share many remarkable similarities in structure, metabolic roles and biosynthesis, indicating their differences are primarily in the minds and economies of man, not the perspectives of producing organisms. Antibiotic/mycotoxin biosynthetic gene packages appear to have been assembled by transposon-mediated processes combining genes acquired horizontally from plants and other soil microbes with genes from the producing organism’s own genome. Probable gene sources include those for the synthesis and secretion of plant phytoalexins, toxins, allelochemicals, pheromones, germination inhibitors and pigments, and bacterial quorum-sensors and siderophores. Understanding antibiotic origins may assist in the discovery and creation of new antibiotics.
    No preview · Article · Jan 2011 · Toxin Reviews
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Bacteria have long been used for the synthesis of a wide range of useful proteins and compounds. The developments of new bioprocesses and improvements of existing strategies for syntheses of valuable products in various bacterial cell hosts have their own challenges and limitations. The field of synthetic biology has combined knowledge from different science and engineering disciplines and facilitated the advancement of novel biological components which has inspired the design of targeted biosynthesis. Here we discuss recent advances in synthetic biology with relevance to biosynthesis in bacteria and the applications of computational algorithms and tools for manipulation of cellular components. Continuous improvements are necessary to keep up with increasing demands in terms of complexity, scale, and predictability of biosynthesis products.
    Full-text · Article · Feb 2012 · FEBS letters
Show more