Significance Microbes have the genetic capacity to produce large numbers of specialized compounds, yet only a small fraction these in laboratory. Here we introduce platform that allows efficient production natural product molecules from uncharacterized gene collections. We used transformation-associated recombination yeast directly clone and express an orphan biosynthetic cluster for lipopeptide antibiotic taromycin A. With this direct cloning approach, single genomic capture expression...

Abstract Bacilli are ubiquitous low G+C environmental Gram-positive bacteria that produce a wide assortment of specialized small molecules. Although their natural product biosynthetic potential is high, robust molecular tools to support the heterologous expression large gene clusters in Bacillus hosts rare. Herein we adapt transformation-associated recombination (TAR) yeast design single genomic capture and vector for antibiotic production subtilis . After validating this direct cloning...

Epsilon-poly-L-lysine (epsilon-PL) is produced by Streptomyces albulus NBRC14147 as a secondary metabolite and can be detected only when the fermentation broth has an acidic pH during stationary growth phase. Since strain produces epsilon-PL-degrading enzymes, original chain length of epsilon-PL polymer product synthesized synthetase (Pls) unclear. Here, we report on identification gene encoding enzyme (PldII), which plays central role in degradation this strain. A knockout mutant pldII was...

Axially chiral biaryl compounds are frequently encountered in nature where they exhibit diverse biological properties. Many biphenols that have C–C or C–O linkages installed by cytochrome P450 oxygenases control the regio- and stereoselectivity of intermolecular coupling reaction. In contrast, bipyrrole-coupling enzymology has not been observed. Marinopyrroles, produced a marine-derived streptomycete, first 1,3′-bipyrrole natural products. On basis marinopyrrole's unusual bipyrrole...

Cyanosporasides are marine bacterial natural products containing a chlorinated cyclopenta[a]indene core of suspected enediyne polyketide biosynthetic origin. Herein, we report the isolation and characterization novel cyanosporasides C–F (3–6) from actinomycetes Salinispora pacifica CNS-143 Streptomyces sp. CNT-179, highlighted by unprecedented C-2′ N-acetylcysteamine functionalized hexose group 6. Cloning, sequencing, mutagenesis homologous ∼50 kb cyanosporaside gene clusters both bacteria...

Abstract Salinamide A belongs to a rare class of bicyclic depsipeptide antibiotics in which the installation (4‐methylhexa‐2,4‐dienoyl)glycine handle across hexadepsipeptide core contributes its chemical complexity and biological properties. Herein, we report genetic biochemical basis for salinamide construction marine bacterium Streptomyces sp. CNB‐091, involves novel intermolecular transesterification reaction catalyzed by type I thioesterase. Heterologous expression studies revealed...

Hyaluronic acid (HA) is used in a wide range of medical applications, where its performance and therapeutic efficacy are highly dependent on molecular weight. In the microbial production HA, it has been suggested that high level intracellular ATP enhances productivity weight HA. Here, we report heterologous HA an ε-poly-l-lysine producer, Streptomyces albulus, which potential to generate at level. The hasA gene from Streptococcus zooepidemicus, encodes synthase, was refactored expressed...

ε-Poly-l-lysine (ε-PL), consisting of 25 to 35 l-lysine residues with linkages between the α-carboxyl groups and ε-amino groups, is produced by Streptomyces albulus NBRC14147. ε-PL synthetase (Pls) a membrane protein six transmembrane domains (TM1 TM6) as well both an adenylation domain thiolation domain, characteristic nonribosomal peptide synthetases. Pls directly generates chain length diversity (25- 35-mer), but processes that control during polymerization reaction are still not fully...

D-Galactose, which did not serve as a growth substrate, was found to induce an L-arabinose isomerase of similar properties the L-arabinose-induced isomerase. In both cases pH profiles, stability, optimum temperature, heat substrate specificity, metal ion requirements, mobility on polyacrylamide gel electrophoresis, and kinetic induced isomerases were identical. It appears possible that D-galactose incorporated into cells by permease system alos D-galactose.

Bacteria produce polycationic homopoly(amino acid)s, which are characterized by isopeptide backbones. Although the biological significance of acid)s remains unclear, increasing attention has recently been focused on their potential use to achieve cellular internalization. Here, for first time, we provide direct evidence that two representative bacterial isopeptides, ε-poly-L-α-lysine (ε-PαL) and ε-oligo-L-β-lysine (ε-OβL), were internalized into mammalian cells cell-membrane penetration then...

The induction of D-xylose, D-ribose, L-arabinose, and D-lyxose isomerases by various sugars was studied to determine the configuration necessary for induction. D-Xylose isomerase only induced whereas D-ribose L-rhamnose, L-lyxose. L-arabinose D-galactose, L-arabitol, D-fucose, dulcitol, D-lyxose, D-mannose, myoinositol. Some compounds such as D- or L-fucose which do not support growth are still able serve inducers pentose isomerases.

Abstract Salinamide A belongs to a rare class of bicyclic depsipeptide antibiotics in which the installation (4‐methylhexa‐2,4‐dienoyl)glycine handle across hexadepsipeptide core contributes its chemical complexity and biological properties. Herein, we report genetic biochemical basis for salinamide construction marine bacterium Streptomyces sp. CNB‐091, involves novel intermolecular transesterification reaction catalyzed by type I thioesterase. Heterologous expression studies revealed...

Among the four bioactive cationic homo-poly(amino acids) discovered in nature, two are mirror-image isomers of poly(2,4-diaminobutyric acid) (poly-Dab) whose biosynthesis has long been unexplained. Their structural analogy plausibly suggested that they could share a common biosynthetic pathway utilizing ε-poly(l-lysine) synthetase-like enzymology but with an unprecedented process for enantiomeric inversion polymer building blocks. To investigate this possibility, we comparatively explored...

Carpel suppression is a trigger for sexual dimorphism in the dioecious plant Silene latifolia. To clarify what kind of genes are involved carpel this species, we generated bisexual mutant, R025, by C-ion beam irradiation. R025 produces flowers with mature gynoecium and stamens. Genetic analysis attributed trait to mutations on Y chromosome. Scanning electron microscopy (SEM) early floral development revealed that size was different from wild-type males spite male background R025. We also...

We found the occurrence of valine dehydrogenase in cell extract a psychrophilic bacterium, Cytophaga sp. KUC-1, isolated from Antarctic seawater and purified enzyme to homogeneity. The molecular mass was determined be approximately 154 kDa by gel filtration that subunit 43 SDS/PAGE: homotetramer. required NAD+ as coenzyme, catalyzed oxidative deamination L-valine, L-isoleucine, L-leucine reductive amination alpha-ketoisovalerate, alpha-ketovalerate, alpha-ketoisocaproate, alpha-ketocaproate....