• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.561-565
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• 2018

The late-stage doxorubicin biosynthesis pathway acting enzyme (DoxA) from Streptomyces peucetius CYP129A2 exhibited substrate promiscuity towards the stilbene group of compounds such as resveratrol. DoxA along with two accessory enzymes ferrdoxin reductase and ferredoxin from spinach hydroxylated resveratrol at the 3'-position in vitro to produce piceatannol. The product was identified by HPLC-PDA and high-resolution HR-qTOF-ESI/MS analyses in positive mode. The ESI/MS fragments resembled the hydroxylated product of resveratrol.

• Bulletin of the Korean Chemical Society
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• 제20권5호
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• pp.543-546
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• 1999

Heme assignment of the 1HNMR spectrum of cytochrome c3 of D. vulgaris Miyazaki F was established [Reference: 12, 13]. The major reduction of the heme turned out to take place in the other of heme 4, 1, 2 and 3 (in the sequential numbering). The Hemes with the smallest and greatest solvent accessibility were reduced at the highest and lowest potentials in average, respectively. A cooperation interheme interaction was attributed to a pait of the closest hemes, namely, hemes 1 and 2. This assignment can provide the physicochemical bases for the elucidation of electron transfer of this protein.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1200-1210
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• 2004

Metabolic flux analysis was established by adapting previous stoichiometric model developed during growth with cellulose to cell grown with cellobiose for further direct comparison of the bacterial metabolism. In carbon limitation with cellobiose, a shift from acetate-ethanol fermentation to ethanol-lactate fermentation is observed and the pyruvate overflow is much higher than with cellulose. In nitrogen limitation with cellobiose, the cellodextrin and exopolysaccharide overflows are much higher than on cellulose. In carbon and nitrogen saturation with cellobiose, the cellodextrin, exopolysaccharide, and free amino acids overflows reach the highest levels observed but all remain limited on cellulose. By completely shunting the cellulosome, the use of cellobiose allows to reach much higher carbon consumption rates which, in return, highlights the metabolic limitation of C. cellulolyticum. Therefore, the physical nature of the carbon source has a profound impact on the metabolism of C. cellulolyticum and most probably of other cellulolytic bacteria. For cellulolytic bacteria, the use of soluble carbon substrate must carefully be taken into consideration for the interpretation of results. Direct comparison of metabolic flux analysis from cellobiose and cellulose revealed the importance of cellulosome, phosphoglucomutase and pyruvate-ferredoxin oxidoreductase in the distribution of carbon flow in the central metabolism. In the light of these findings, future directions for improvement of cellulose catabolism by this bacterium are discussed.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.237-243
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• 2020

A novel psychrotolerant Psychrobacillus strain PB01, isolated from an Antarctic iceberg, was comparatively analyzed with five related strains. The complete genome of strain PB01 consists of a single circular chromosome (4.3 Mb) and a plasmid (19 Kb). As potential low-temperature adaptation strategies, strain PB01 has four genes encoding cold-shock proteins, two genes encoding DEAD-box RNA helicases, and eight genes encoding transporters for glycine betaine, which can serve as a cryoprotectant, on the genome. The pan-genome structure of the six Psychrobacillus strains suggests that strain PB01 might have evolved to adapt to extreme environments by changing its genome content to gain higher capacity for DNA repair, translation, and membrane transport. Notably, strain PB01 possesses a complete TCA cycle consisting of eight enzymes as well as three additional Helicobacter pylori-type enzymes: ferredoxin-dependent 2-oxoglutarate synthase, succinyl-CoA/acetoacetyl-CoA transferase, and malate/quinone oxidoreductase. The co-existence of the genes for TCA cycle enzymes has also been identified in the other five Psychrobacillus strains.

• BMB Reports
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• 제45권4호
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• pp.239-241
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• 2012

Iron availability is limited in the environment and most bacteria have developed a system to acquire iron from host hemoproteins. Heme oxygenase plays an important role by degrading heme group and releasing the essential nutrient iron. The structure of Bacillus subtilis HmoB was determined to 2.0 ${\AA}$ resolution. B. subtilis HmoB contains a typical antibiotic biosynthesis monooxygenase (ABM) domain that spans from 71 to 146 residues and belongs to the IsdG family heme oxygenases. Comparison of HmoB and IsdG family proteins showed that the C-terminal region of HmoB has similar sequence and structure to IsdG family proteins and contains conserved critical residues for heme degradation. However, HmoB is distinct from other IsdG family proteins in that HmoB is about 60 amino acids longer in the N-terminus and does not form a dimer whereas previously studied IsdG family heme oxygenases form functional homodimers. Interestingly, the structure of monomeric HmoB resembles the dimeric structure of IsdG family proteins. Hence, B. subtilis HmoB is a heme oxygenase with a novel structural feature.

• Journal of Microbiology and Biotechnology
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• 제16권2호
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• pp.295-298
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• 2006

Actinomycetes are ubiquitous Gram-positive soil bacteria and a group of the most important industrial microorganisms for the biosynthesis of many valuable secondary metabolites as well as the source of various bioconversion enzymes. Cytochrome P450 hydroxylase (CYP), a hemebinding protein, is known to be involved in the modification of various natural compounds, including polyketides, fatty acids, steroids, and some aromatic compounds. Previously, six different novel CYP genes were isolated from a rare actinomycetes called Sebekia benihana, and they were completely sequenced, revealing significant amino acid similarities to previously known CYP genes involved in Streptomyces secondary metabolism. In the present study, these six CYP genes were functionally expressed in Streptomyces lividans, using an $ermE^{*}$ promoter-containing Streptomyces expression vector. Among six CYP genes, two S. benihana CYP genes (CYP503 and CYP504) showed strong hydroxylation activities toward 7-ethoxycoumarin. Furthermore, the recombinant S. lividans containing both the S. benihana CYP506-ferredoxin genes as well as the S. coelicolor feredoxin reductase gene also demonstrated cyclosporin A hydroxylation activity, suggesting potential application of actinomycetes CYPs for the biocatalysts of natural product bioconversion.

• Journal of Photoscience
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• 제3권2호
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• pp.77-83
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• 1996

Photosensitization via the singlet oxygen ($^1O_2$) mechanism by the binuclear iron-sulfur center, denoted as [2Fe-2S], was investigated, using a highly purified ferredoxin (Fd) preparation from spinach leaves. Since the apoprotein of Fd contains a good number of amino acid residues that are readily reactive with $^1O_2$ and thus interfere with the detection of $^1O_2$ generated from [2Fe-2S], we attempted to deprive the $^1O_2$-sensitive residues of their $^1O_2$-scavenging capacity as much as possible by treating Fd with rose bengal plus 550 nm monochromatic light and thereby photooxidatively degrading these residues. The photochemically modified Fd was found to keep the structural integrity of its Fe-S group virtually unaffected by the treatment. By employing chemical trap method for measurement and examining the kinetic effects of azide and deuterium oxide on the reactions of $^1O_2$ with various trap compounds, we were able to demonstrate that [2Fe-2S] indeed acts as a photosensitizer via $^1O_2$. Further, the minimum quantum yield of $^1O_2$ production by [2Fe-2S] was estimated to be 0.0047.

• The Korean Journal of Ecology
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• 제14권1호
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• pp.87-99
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• 1991

Changes of phycobiliproteins(PBP) quantity and ferredoxin-NADP reductase(FNR) activity were investigated in various light illuminated cyanobacteria, Anabaena variabilis. PBP components were increased under blue light illumination, whereas decreased under red light illumination. PBP contents were twofolds in blue light than in red light. In view of the PBP composition, allophycocyanin(APC) in red light was higher 5.5% and phycoerythrocyanin(PEC) in blue light was higher 2.2% than in white light-illuminated PBP. It was suggested that PBP changes in bule light be the results of regulation of photosysthetic efficiency and protection of photosystem, whereas PBP changes in red light be effected by adaptation of adequate harvesting of light energy in photosystem. Changes of FNR activity were highest in red light, and sequenced lower to blue light and green light. It means that light-dependent production rate of NADP is the highest in red light. The difference of values was larger than that of values in comparison of red and blue light. It was suggested that increasing of FNR activity be due not to the function of isozyme, but to the synthesis of enzymes. Because of NAD/NADP regulation-effect to metabolism, it was considered that FNR activity might influence the metabolism indirectly and explain the probability of regulation in pathways of key enzyme activation. FNR activity was directly proportional to intensity of light. Optimum temperature and pH were about 25℃ and 7.5, respectively.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1996년도 식물학심포지움 식물호르몬과 신호전달
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• pp.50-63
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• 1996

Nitrogen (N) is an important regulator of the expression of genes involved in carbon and N assimilation pathways in plants by selectively altering the levels of proteins and/or mRNAs. These in C4 plants include genes for such as phosphoenolpyruvate carboxylase, carbonic anhydrase, and pyruvate-Pi dikinase. The C4 genes are regulated in mesophyll cells by N availability both transcriptionally and posttranscriptionally through cytokinins and glutamine as signals. The level of both the signals is up-regulated by N availability: cytokinins in roots and glutamine in leaves. The level of glutamine is controlled by the differential expression by N of glutamine synthetase and ferrdoxin-dependent glutamate synthase genes which locate in the mesophyll cells of C4 plants. The results is discussed as molecular mechanism for the greater N use efficiency of the plants as well as N partitioning is the photosynthetic cells.

• 한국자기공명학회논문지
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• 제17권2호
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• pp.105-110
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• 2013

HP1492 is a NifU-like protein of Helicobacter pylori (H. pylori) and plays a role as a scaffold which transfer Fe-S cluster to Fe-S proteins like Ferredoxin. To understand how to bind to iron ion or iron-sulfur cluster, HP1492 was expressed and purified in Escherichia coli (E. coli). From the NMR measurement, we could carry out the sequence specific backbone resonance assignment of HP1492. Approximately 91% of all resonances could be assigned unambiguously. By analyzing results of CSI and TALOS from NMR data, we could predict the secondary structure of HP1492, which consists of three ${\alpha}$-helices and three ${\beta}$-sheets. This study is an essential step towards the structural characterization of HP1492.