The macrocyclic core of the antibiotic erythromycin, 6-deoxyerythronolide B (6dEB), is a complex natural product synthesized by soil bacterium Saccharopolyspora erythraea through action multifunctional polyketide synthase (PKS). engineering potential modular PKSs hampered limited capabilities for molecular biological manipulation organisms (principally actinomycetes) in which polyketides have thus far been produced. To address this problem, derivative Escherichia coli has genetically...

Significance Terpenes are generally considered to be plant or fungal metabolites, although a small number of odoriferous terpenes bacterial origin have been known for many years. Recently, extensive genome sequencing and bioinformatic analysis deduced proteins using profile based on hidden Markov model revealed 262 distinct predicted terpene synthases. Although these presumptive synthase genes seem silent in their parent microorganisms, controlled expression an engineered heterologous...

To construct a versatile model host for heterologous expression of genes encoding secondary metabolite biosynthesis, the genome industrial microorganism Streptomyces avermitilis was systematically deleted to remove nonessential genes. A region more than 1.4 Mb stepwise from 9.02-Mb S. linear chromosome generate series defined deletion mutants, corresponding 83.12–81.46% wild-type chromosome, that did not produce any major endogenous metabolites found in parent strain. The suitability mutants...

The crystal structure of pentalenene synthase at 2.6 angstrom resolution reveals critical active site features responsible for the cyclization farnesyl diphosphate into tricyclic hydrocarbon pentalenene. Metal-triggered substrate ionization initiates catalysis, and α-barrel serves as a template to channel stabilize conformations reactive carbocation intermediates through complex cascade. core enzyme may be preserved among greater family terpenoid synthases, possibly implying divergence from...

An industrial microorganism, Streptomyces avermitilis, which is a producer of anthelmintic macrocyclic lactones, avermectins, has been constructed as versatile model host for heterologous expression genes encoding secondary metabolite biosynthesis. Twenty the entire biosynthetic gene clusters metabolites were successively cloned and introduced into S. avermitilis SUKA17 or 22. Almost all transformants carrying cluster produced result introduced. A few unable to produce metabolites, but their...

Modular polyketide synthases catalyze the biosynthesis of medicinally important natural products through an assembly-line mechanism. Although these megasynthases display very precise overall selectivity, we show that their constituent modules are remarkably tolerant toward diverse incoming acyl chains. By appropriate engineering linkers, which exist within and between polypeptides, it is possible to exploit this tolerance facilitate transfer biosynthetic intermediates unnaturally linked...

The x-ray crystal structure of a 194-kDa fragment from module 5 the 6-deoxyerythronolide B synthase has been solved at 2.7 Å resolution. Each subunit homodimeric protein contains full-length ketosynthase (KS) and acyl transferase (AT) domain as well three flanking “linkers.” linkers are structurally defined contribute extensively to intersubunit or interdomain interactions, frequently by means multiple highly conserved residues. also reveals that active site residue Cys-199 KS is separated...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTBiosynthetic origin of the carbon skeleton and oxygen atoms nargenicin A1David E. Cane Chi Ching YangCite this: J. Am. Chem. Soc. 1984, 106, 3, 784–787Publication Date (Print):February 1, 1984Publication History Published online1 May 2002Published inissue 1 February 1984https://pubs.acs.org/doi/10.1021/ja00315a052https://doi.org/10.1021/ja00315a052research-articleACS PublicationsRequest reuse permissionsArticle Views185Altmetric-Citations250LEARN...

The x-ray crystal structure of recombinant trichodiene synthase from Fusarium sporotrichioides has been determined to 2.5-Å resolution, both unliganded and complexed with inorganic pyrophosphate. This reaction product coordinates three Mg 2+ ions near the mouth active site cleft. A comparison liganded structures reveals a ligand-induced conformational change that closes Binding substrate farnesyl diphosphate similarly may trigger this change, which would facilitate catalysis by protecting...

A genetic block was introduced in the first condensation step of polyketide biosynthetic pathway that leads to formation 6-deoxyerythronolide B (6-dEB), macrocyclic precursor erythromycin. Exogenous addition designed synthetic molecules small-scale cultures this null mutant resulted highly selective multimilligram production unnatural polyketides, including aromatic and ring-expanded variants 6-dEB. Unexpected incorporation patterns were observed, illustrating catalytic versatility modular...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTUnified stereochemical model of polyether antibiotic structure and biogenesisDavid E. Cane, Walter D. Celmer, John W. WestleyCite this: J. Am. Chem. Soc. 1983, 105, 11, 3594–3600Publication Date (Print):June 1, 1983Publication History Published online1 May 2002Published inissue 1 June 1983https://pubs.acs.org/doi/10.1021/ja00349a040https://doi.org/10.1021/ja00349a040research-articleACS PublicationsRequest reuse permissionsArticle...

As the first structural elucidation of a modular polyketide synthase (PKS) domain, crystal structure macrocycle-forming thioesterase (TE) domain from 6-deoxyerythronolide B (DEBS) was solved by combination multiple isomorphous replacement and multiwavelength anomalous dispersion refined to an R factor 24.1% 2.8-Å resolution. Its overall tertiary architecture belongs α/β-hydrolase family, with two unusual features unprecedented in this family: hydrophobic leucine-rich dimer interface...

The fungus Botrytis cinerea is the causal agent of economically important gray mold disease that affects more than 200 ornamental and agriculturally plant species. B. a necrotrophic pathogen secretes nonspecific phytotoxins, including sesquiterpene botrydial polyketide botcinic acid. region surrounding previously characterized BcBOT1 gene has now been identified as biosynthetic cluster. Five genes BcBOT2 were shown by quantitative reverse transcription-PCR to be co-regulated through...

Cytochrome P450 170A1 (CYP170A1) is encoded by the sco5223 gene of Gram-positive, soil-dwelling bacterium Streptomyces coelicolor A3(2) as part a two-gene cluster with sco5222 gene. The SCO5222 protein sesquiterpene synthase that catalyzes cyclization farnesyl diphosphate to novel tricyclic hydrocarbon, epi-isozizaene (Lin, X., Hopson, R., and Cane, D. E. (2006) J. Am. Chem. Soc. 128, 6022-6023). presence CYP170A1 (sco5223) suggested epiisozizaene might be further oxidized transcriptionally...

The X-ray crystal structure of recombinant epi-isozizaene synthase (EIZS), a sesquiterpene cyclase from Streptomyces coelicolor A3(2), has been determined at 1.60 Å resolution. Specifically, the wild-type EIZS is that its closed conformation in complex with three Mg2+ ions, inorganic pyrophosphate (PPi), and benzyltriethylammonium cation (BTAC). Additionally, D99N an open, ligand-free 1.90 Comparison these two structures provides first view conformational changes required for substrate...

Methylisoborneol (2) is a volatile organic compound produced by wide variety of Actinomycete soil organisms, myxobacteria, and cyanobacteria. It has an unusually low odor threshold and, together with geosmin, responsible for the characteristic smell moist as well unpleasant taste episodes associated public water supplies contamination various foodstuffs, including fish, wine, beer. Despite considerable interest in detection remediation methylisoborneol, biosynthesis this methylated...