The gene locus for acetyl-CoA carboxylase (ACC) involved in the primary metabolism was identified from the genomic library of Streptomyces toxytricini which produces a lipase inhibitor lipstatin. The 7.4 kb cloned gene was comprised of 5 ORFs including accD1, accA1, hmgL, fadST1, and stsF. In order to confirm the biochemical characteristics of AccA1, the gene was overexpressed in Escherichia coli cells, and the recombinant protein was purified through Ni2+ affinity chromatography. Because most of the expressed AccAl was biotinylated by host E. coli BirA in the presence of D-biotin, the non-biotinylated apo-AccA1 was purified after gene induction without D-biotin, followed by exclusion of holo-AccA1 using streptavidin beads. The separated apo-AccA1 was post-translationally biotinylated by S. toxytricini biotin apo-protein ligase (BPL) in a time- and enzyme-dependent manner. This result supports that this gene cluster of S. toxytricini encodes the functional ACC enzyme subunits to be biotinylated.
Streptomyces toxytricini produces lipstatin, a specific inhibitor of pancreatic lipase, which is derived from two fatty acid moieties with eight and 14 carbon atoms. The pccB gene locus in 10.6 kb fragment of S. toxytricini chromosomal DNA contains three genes for acyl-coenzyme A carboxylase (ACCase) complex accA3, pccB, and pccE that are presumed to be involved in secondary metabolism. The pccB gene encoding a β subunit of ACCase [carboxyltransferase (CT)] was identified upstream of pccE gene for a small protein of ε subunit. The accA3 encoding the α subunit of ACCase [biotin carboxylase (BC)] was also identified downstream of pccB gene. When the pccB and pccE genes were inactivated by homologous recombination, the lipstatin production was reduced as much as 80%. In contrast, the accumulation of another compound, tetradeca-5.8-dienoic acid (the major lipstatin precursor), was 4.5-fold increased in disruptant compared with wild-type. It implies that PccB of S. toxytricini is involved in the activation of octanoic acid to hexylmalonic acid for lipstatin biosynthesis.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-010-2587-2) contains supplementary material, which is available to authorized users.
Deep blue emitting copolymers were synthesized by uniting the Eumelanin-inspired indole core with fluorene and carbazole units via Suzuki polymerization. The resulting polymers, PIF and PIC, showed deep blue emission in the range of 416-418 nm and quantum yields of 0.39-0.60. Both polymers exhibited an intense and stable electrogenerated chemiluminescence. Interestingly, deep HOMO levels of 25.71 and 25.61 eV were observed for PIF and PIC, respectively. Solution processed polymer light emitting diodes (PLEDs) were fabricated using the PIF as a guest. PLEDs emitted deep blue light at 418 nm, with the luminous efficiency peaking at 1 Cd/A, given that the photopic response at that wavelength is 0.0151. The electroluminescence of PIF displayed a Commission Internationale de l'Eclairage coordinates of (0.16, 0.07) with a maximum external quantum efficiency of 1.1%. Hence, these materials prove to be promising candidates for the fabrication of deep blue PLEDs.
Four novel conjugated polymers containing the eumelanin-inspired indole core have been successfully synthesized using common cross coupling reactions. These polymers differed by the arylene and the carbon-carbon bond linkage. Optoelectronic experiments of these polymers suggest that the ethynylene linkage contributed to the red-shifted absorption spectra and blue-shifted emission spectra when compared to the vinylene linkage polymers. Furthermore, the optical bandgaps of the poly(indoylenearyleneethynylene)s (PIAEs) were smaller compared to the poly(indoylenearylenevinylene)s (PIAVs). Surprisingly, the HOMOs of these polymers were less affected by the nature of the carbon-carbon linkage. However, the LUMOs of the PIAEs were lower in comparison to the PIAVs. These eumelanin-inspired PIAEs and PIAVs are good fluorophores with fluorescence quantum yields ranging from 0.12 to 0.67 and have good thermal stability for applications such as in organic light-emitting diodes.
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