• 한국당과학회:학술대회논문집
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• 2006.11a
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• pp.52-52
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• 2006

• 한국당과학회:학술대회논문집
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• 2006.11a
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• pp.51-51
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• 2006

• Molecules and Cells
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• v.20 no.1
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• pp.90-96
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• 2005

A cluster of genes for ribostamycin (Rbm) biosynthesis was isolated from Streptomyces ribosidificus ATCC 21294. Sequencing of 31.892 kb of the genomic DNA of S. ribosidificus revealed 26 open reading frames (ORFs) encoding putative Rbm biosynthetic genes as well as resistance and other genes. One of ten putative Rbm biosynthetic genes, rbmA, was expressed in S. lividans TK24, and shown to encode 2-deoxy-scyllo-inosose (DOI) synthase. Acetylation of various aminoglycoside-aminocyclitol (AmAcs) by RbmI confirmed it to be an aminoglycoside 3-N-acetyltransferase. Comparison of the genetic control of ribostamycin and butirosin biosynthesis pointed to a common biosynthetic route for these compounds, despite the considerable differences between them in genetic organization.

• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.828-831
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• 2003

Although most of the DOS containing aminoglycosides are produced by Streptomyces, very little information is available about their biosynthesis. In the present paper, we report a method to isolate DOI synthase, a key enzyme in the biosynthesis of DOS, from aminoglycoside producer Streptomyces. PCR primers based on conserved region of DOI synthases were specific and reliable for the isolation of the biosynthetic genes of DOS containing aminoglycosides or the screening of the aminoglycoside producers. The use of DOI synthase as a probe could save both time and cost of cloning aminoglycoside biosynthetic genes.

• Proceedings of the PSK Conference
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• 2003.10a
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• pp.81-84
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• 2003

Aminoglycoside antibiotics are among the chlinically important antibiotics and a major structural feature is the existence of characteristic aminocyclitol aglycones [1]. Considering the structural features, aminoglycosides are classified in to two classes. The first are those containing fully substituted aminocyclitol such as streptomycin, hygromycin, fortimycin, etc. (omitted)

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.116-120
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• 2006

We constructed four recombinant plasm ids to enhance the production of clavulanic acid (CA) in Streptomyces clavuligerus NRRL3585: (1) pIBRHL1, which includes ccaR, a pathway-specific regulatory gene involved in cephamycin C and CA biosynthesis; (2) pIBRHL2, containing claR, again a regulatory gene, which controls the late steps of CA biosynthesis; (3) pGIBR containing afsR-p, a global regulatory gene from Streptomyces peucetius; and (4) pKS, which harbors all of the genes (ccaR/ claR/ afsR-p). The plasmids were expressed in S. clavuligerus NRRL3585 along with the $ermE^*$ promoter. All of them enhanced the production of CA; 2.5-fold overproduction for pIBRHL1, 1.5-fold for pIBRHL2, 1.6-fold for pGIBR, and 1.5-fold for pKS compared to the wild type.

• The Transactions of The Korean Institute of Electrical Engineers
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• v.58 no.1
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• pp.147-154
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• 2009

This paper presents a grating~integrated SPR (Surface Plasmon Resonance) sensor chip for simple and inexpensive biomolecule detection. The grating-integrated SPR sensor chip has two sensing channels having a nano grating for SPR coupling. An external mirror is used for multi channel SPR sensing. The present sensor chip replaces bulky and expensive optical components, such as fiber-optic switches or special shaped prisms, resulting in a simple and inexpensive wavelength modulated multi-channel SPR sensing system. We fabricate a SPR sensor chip integrated with 835 nm-pitch gratings by a micromolding technique to reduce the fabrication cost. In the experimental characterization, the refractive index sensitivity of each sensing channel is measured as $321.8{\pm}8.1nm$/RI and $514.3{\pm}8.lnm$/RI, respectively. 0.5uM of the target biomolecule (streptavidin) was detected by a $1.13{\pm}0.16nm$ shift of the SPR dip in the 10%-biotinylated sample channel, while the SPR dip in the reference channel for environmental perturbation monitoring remained at the same position. From the experimental results, multi-channel biomolecule detection capability of the present grating-integrated SPR sensor chip has been verified. On the basis of the preliminary experiments, we successfully measured the binding reaction rate for the $2\;nM{\sim}200\;nM$ monoclonal-antibiotin, thus verifying biomolecule concentration detectability of the present SPR sensor chip. The binding reaction rates measured from the present SPR sensor chip agredd well with those from a commercialized SPR sensor.

• BMB Reports
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• v.30 no.5
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• pp.337-341
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• 1997

In order to investigate the role of a small GTP-binding protein RhoA in lysophosphatidic acid (LPA)-induced stress fiber formation, C3 ADP-ribosyltransferase was prepared by expressing in E. coli and then applied to Rat-2 fibroblasts. C3 transferase isolated from E. coli was as effective as the toxin from Clostridium botulinum in ADP-ribosylation of RhoA. Incubation of the cells with C3 transferase for 2 days induced ADP-ribosylation of RhoA by a dose-dependent manner, with a sub-maximal induction at $25\;{\mu}g/ml$. As expected, LPA-induced stress fiber formation was completely blocked by pre-incubation with C3 transferase for 2 days. However, exogenously added C3 transferase had no significant effect on the formation of phosphatidylethanol by LPA. These results suggested that phospholipase D was not activated by RhoA in the LPA-induced stress fiber formation.

• Journal of Microbiology and Biotechnology
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• v.16 no.12
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• pp.1928-1934
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• 2006

The rubradirin biosynthetic gene cluster harbors 58 ORFs within a 105.6-kb sequence, which includes all of the genes responsible for the synthesis of rubradirin, as well as the primary genes relevant to regulatory, resistance, and transport functions. This gene cluster also harbors a resistance-mediating ABC transporter, RubT1, which is located at the most upstream position in the cluster. In the present study, RubT1 was expressed heterologously in E. coli, and the resistance affinity of RubT1 was determined by an antibacterial activity test, as well as by HPLC and ESI-MS analyses. Evidence clearly demonstrates that RubTl mediates rubradirin resistance as an ABC transporter.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1059-1065
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• 2012

Biocatalytic transfer of oxygen in isolated cytochrome P450 or whole microbial cells is an elegant and efficient way to achieve selective hydroxylation. Cytochrome P450 CYP105P2 was isolated from Streptomyces peucetius that showed a high degree of amino acid identity with hydroxylases. Previously performed homology modeling, and subsequent docking of the model with flavone, displayed a reasonable docked structure. Therefore, in this study, in a pursuit to hydroxylate the flavone ring, CYP105P2 was co-expressed in a two-vector system with putidaredoxin reductase (camA) and putidaredoxin (camB) from Pseudomonas putida for efficient electron transport. HPLC analysis of the isolated product, together with LC-MS analysis, showed a monohydroxylated flavone, which was further established by subsequent ESI/MS-MS. A successful 10.35% yield was achieved with the whole-cell bioconversion reaction in Escherichia coli. We verified that CYP105P2 is a potential bacterial hydroxylase.