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Atanas V. Demirev Ji Seon Lee Bhishma R. Sedai Ivan G. Ivanov Doo Hyun Nam 《Journal of microbiology (Seoul, Korea)》2009,47(4):473-478
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. 相似文献
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Atanas V. Demirev Anamika Khanal Bhishma R. Sedai Si Kyu Lim Min Kyun Na Doo Hyun Nam 《Applied microbiology and biotechnology》2010,87(3):1129-1139
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. 相似文献
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Rat Leydig cells were permeabilized and the cytoplasm partially extracted to visualize, describe, and characterize filamentous elements of the cytoskeleton. It was demonstrated by immunofluorescence microscopy that vimentin is abundant within Leydig cells. Ultrastructurally, intermediate filaments in Leydig cells were concentrated at perinuclear sites and comprised bundles that coursed through the cytoplasm. Actin was identified in Leydig cells with the F actin probe, NBD-phallacidin. Fluorescence was strongest at the cortex of the cell. With myosin S-1 subfragments, sparse actin was found positioned almost exclusively in cortical regions of the cell associated with coated pits and in Leydig cell processes. 相似文献
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Karki Bhishma Sharma Sandeep Singh Yadvendra Pal Amrindra 《Plasmonics (Norwell, Mass.)》2022,17(1):71-78
Plasmonics - The sensitivity of a standard surface plasmon resonance (SPR) biosensor in angular interrogation is low. A unique 2-dimensional substantial heterostructure, franckeite/graphene layer,... 相似文献
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