| inactivation, complementation, and heterologous expression of encp, a novel bacterial phenylalanine ammonia-lyase gene. | the enzyme phenylalanine ammonia-lyase, which catalyzes the nonoxidative deamination of l-phenylalanine to trans-cinnamic acid, is ubiquitously distributed in plants. we now report its characterization for the first time in a bacterium. the phenylalanine ammonia-lyase homologous gene encp from the "streptomyces maritimus" enterocin biosynthetic gene cluster was functionally characterized and shown to encode the first enzyme in the pathway to the enterocin polyketide synthase starter unit benzoyl ... | 2002 | 12082112 |
| molecular cloning and sequence analysis of the clorobiocin biosynthetic gene cluster: new insights into the biosynthesis of aminocoumarin antibiotics. | the biosynthetic gene cluster of the aminocoumarin antibiotic clorobiocin was cloned by screening of a cosmid library of streptomyces roseochromogenes ds 12.976 with two heterologous probes from the novobiocin biosynthetic gene cluster. sequence analysis revealed 27 orfs with striking similarity to the biosynthetic gene clusters of novobiocin and coumermycin a(1). inactivation of a putative aldolase gene, clor, by in-frame deletion led to the abolishment of the production of clorobiocin. feeding ... | 2002 | 12480894 |
| plant-like biosynthetic pathways in bacteria: from benzoic acid to chalcone. | although phenylpropanoids and flavonoids are common plant natural products, these major classes of biologically active secondary metabolites are largely absent from bacteria. the ubiquitous plant enzymes phenylalanine ammonia-lyase (pal) and chalcone synthase (chs) are key biosynthetic catalysts in phenylpropanoid and flavonoid assembly, respectively. until recently, few bacterial counterparts were known, thus reflecting the dearth of these plant natural products in bacteria. this review highlig ... | 2002 | 12502351 |
| characterization of benzoyl coenzyme a biosynthesis genes in the enterocin-producing bacterium "streptomyces maritimus". | the novel benzoyl coenzyme a (benzoyl-coa) biosynthesis pathway in "streptomyces maritimus" was investigated through a series of target-directed mutations. genes involved in benzoyl-coa formation were disrupted through single-crossover homologous recombination, and the resulting mutants were analyzed for their ability to biosynthesize the benzoyl-coa-primed polyketide antibiotic enterocin. inactivation of the unique phenylalanine ammonia-lyase-encoding gene encp was previously shown to be absolu ... | 2003 | 12511484 |
| mutasynthesis of enterocin and wailupemycin analogues. | inactivation of the novel phenylalanine ammonia lyase gene encp, whose product is a key component in the biosynthetic pathway to benzoyl-coenzyme a (coa) in the bacterium streptomyces maritimus, resulted in the loss of production of the benzoate-primed polyketides enterocin and wailupemycin g. a series of cinnamate and benzoate derivatives were administered to the deltaencp mutant, resulting in the formation of novel analogues bearing p-fluorobenzoate, 2- and 3-thiophenecarboxylate, and cyclohex ... | 2003 | 12889947 |
| context-dependent behavior of the enterocin iterative polyketide synthase; a new model for ketoreduction. | heterologous expression and mutagenesis of the enterocin type ii polyketide synthase (pks) system suggest for the first time that the association of an extended set of proteins and substrates is needed for the effective production of the enterocin-wailupemycin polyketides. in the absence of its endogenous ketoreductase (kr) encd in either the enterocin producer "streptomyces maritimus" or the engineered host s. lividans k4-114, the enterocin minimal pks is unable to produce benzoate-primed polyk ... | 2004 | 15123240 |
| encm, a versatile enterocin biosynthetic enzyme involved in favorskii oxidative rearrangement, aldol condensation, and heterocycle-forming reactions. | the bacteriostatic natural product enterocin from the marine microbe "streptomyces maritimus" has an unprecedented carbon skeleton that is derived from an aromatic polyketide biosynthetic pathway. its caged tricyclic, nonaromatic core is derived from a linear poly-beta-ketide precursor that formally undergoes a favorskii-like oxidative rearrangement. in vivo characterization of the gene encm through mutagenesis and heterologous biosynthesis demonstrated that its protein product not only is solel ... | 2004 | 15505225 |
| biochemical characterization of a prokaryotic phenylalanine ammonia lyase. | the committed biosynthetic reaction to benzoyl-coenzyme a in the marine bacterium "streptomyces maritimus" is carried out by the novel prokaryotic phenylalanine ammonia lyase (pal) encp, which converts the primary amino acid l-phenylalanine to trans-cinnamic acid. recombinant encp is specific for l-phenylalanine and shares many biochemical features with eukaryotic pals, which are substantially larger proteins by approximately 200 amino acid residues. | 2005 | 15937191 |
| the essential tyrosine-containing loop conformation and the role of the c-terminal multi-helix region in eukaryotic phenylalanine ammonia-lyases. | besides the post-translationally cyclizing catalytic ala-ser-gly triad, tyr110 and its equivalents are of the most conserved residues in the active site of phenylalanine ammonia-lyase (pal, ec 4.3.1.5), histidine ammonia-lyase (hal, ec 4.3.1.3) and other related enzymes. the tyr110phe mutation results in the most pronounced inactivation of pal indicating the importance of this residue. the recently published x-ray structures of pal revealed that the tyr110-loop was either missing (for rhodosprid ... | 2006 | 16478474 |
| comparative genomics of streptomyces avermitilis, streptomyces cattleya, streptomyces maritimus and kitasatospora aureofaciens using a streptomyces coelicolor microarray system. | dna/dna microarray hybridization was used to compare the genome content of streptomyces avermitilis, streptomyces cattleya, streptomyces maritimus and kitasatospora aureofaciens with that of streptomyces coelicolor a3(2). the array data showed an about 93% agreement with the genome sequence data available for s. avermitilis and also showed a number of trends in the genome structure for streptomyces and closely related kitasatospora. a core central region was well conserved, which might be predic ... | 2008 | 17588127 |
| enzymatic total synthesis of enterocin polyketides. | polyketides are clinically important natural products that often require elaborate organic syntheses owing to their complex chemical structures. here we report the multienzyme total synthesis of the streptomyces maritimus enterocin and wailupemycin bacteriostatic agents in a single reaction vessel from simple benzoate and malonate substrates. to our knowledge, our results represent the first in vitro assembly of a complete type ii polyketide synthase enzymatic pathway to natural products. | 2007 | 17704772 |
| in vitro biosynthesis of unnatural enterocin and wailupemycin polyketides. | nature has evolved finely tuned strategies to synthesize rare and complex natural products such as the enterocin family of polyketides from the marine bacterium streptomyces maritimus. herein we report the directed ex vivo multienzyme syntheses of 24 unnatural 5-deoxyenterocin and wailupemycin f and g analogues, 18 of which are new. we have generated molecular diversity by priming the enterocin biosynthesis enzymes with unnatural substrates and have illustrated further the uniqueness of this typ ... | 2009 | 19215142 |
| cinnamic acid production using streptomyces lividans expressing phenylalanine ammonia lyase. | cinnamic acid production was demonstrated using streptomyces as a host. a gene encoding phenylalanine ammonia lyase (pal) from streptomyces maritimus was introduced into streptomyces lividans, and its expression was confirmed by western blot analysis. after 4 days cultivation using glucose as carbon source, the maximal level of cinnamic acid reached 210 mg/l. when glycerol (30 g/l) was used as carbon source, the maximal level of produced cinnamic acid reached 450 mg/l. in addition, using raw sta ... | 2011 | 21424686 |
| Production of Streptoverticillium cinnamoneum transglutaminase and cinnamic acid by recombinant Streptomyces lividans cultured on biomass-derived carbon sources. | Transglutaminase from Streptoverticillium cinnamoneum (StvcMTG) was produced using recombinant Streptomyces lividans. When grown on glycerol and xylose as sole carbon sources, S. lividans/StvcMTG produced 360 and 530mg of StvcMTG per liter, respectively. With starch and xylan, the strain produced 230 and 400mg of StvcMTG per liter, respectively. Recombinant S. lividans/encP, which expresses phenylalanine ammonia lyase from Streptomyces maritimus, produced 160mg/L of cinnamic acid from cellulose. ... | 2012 | 22115528 |
| benzoic acid fermentation from starch and cellulose via a plant-like β-oxidation pathway in streptomyces maritimus. | benzoic acid is one of the most useful aromatic compounds. despite its versatility and simple structure, benzoic acid production using microbes has not been reported previously. streptomyces are aerobic, gram-positive, mycelia-forming soil bacteria, and are known to produce various kinds of antibiotics composed of many aromatic residues. s. maritimus possess a complex amino acid modification pathway and can serve as a new platform microbe to produce aromatic building-block compounds. in this stu ... | 2012 | 22545774 |
| direct capture and heterologous expression of salinispora natural product genes for the biosynthesis of enterocin. | heterologous expression of secondary metabolic pathways is a promising approach for the discovery and characterization of bioactive natural products. herein we report the first heterologous expression of a natural product from the model marine actinomycete genus salinispora. using the recently developed method of yeast-mediated transformation-associated recombination for natural product gene clusters, we captured a type ii polyketide synthase pathway from salinispora pacifica with high homology ... | 2015 | 25382643 |
| the bacterial ammonia lyase encp: a tunable biocatalyst for the synthesis of unnatural amino acids. | enzymes of the class i lyase-like family catalyze the asymmetric addition of ammonia to arylacrylates, yielding high value amino acids as products. recent examples include the use of phenylalanine ammonia lyases (pals), either alone or as a gateway to deracemization cascades (giving (s)- or (r)-α-phenylalanine derivatives, respectively), and also eukaryotic phenylalanine aminomutases (pams) for the synthesis of the (r)-β-products. herein, we present the investigation of another family member, en ... | 2015 | 26390197 |
| discovery, biosynthesis, and rational engineering of novel enterocin and wailupemycin polyketide analogues. | the marine actinomycete streptomyces maritimus produces a structurally diverse set of unusual polyketide natural products including the major metabolite enterocin. investigations of enterocin biosynthesis revealed that the unique carbon skeleton is derived from an aromatic polyketide pathway which is genetically coded by the 21.3 kb enc gene cluster in s. maritimus. characterization of the enc biosynthesis gene cluster and subsequent manipulation of it via heterologous expression and/or mutagene ... | 2013 | 23963911 |
| a peroxisomally localized acyl-activating enzyme is required for volatile benzenoid formation in a petuniaxhybrida cv. 'mitchell diploid' flower. | floral volatile benzenoid/phenylpropanoid (fvbp) biosynthesis is a complex and coordinate cellular process executed by petal limb cells of a petunia×hybrida cv. 'mitchell diploid' (md) plant. in md flowers, the majority of benzenoid volatile compounds are derived from a core phenylpropanoid pathway intermediate by a coenzyme a (coa) dependent, β-oxidative scheme. metabolic flux analysis, reverse genetics, and biochemical characterizations of key enzymes in this pathway have supported this putati ... | 2012 | 22771854 |