• Title/Summary/Keyword: Cofactor

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Omega Rhodopsins: A Versatile Class of Microbial Rhodopsins

  • Kwon, Soon-Kyeong;Jun, Sung-Hoon;Kim, Jihyun F.
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.633-641
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    • 2020
  • Microbial rhodopsins are a superfamily of photoactive membrane proteins with the covalently bound retinal cofactor. Isomerization of the retinal chromophore upon absorption of a photon triggers conformational changes of the protein to function as ion pumps or sensors. After the discovery of proteorhodopsin in an uncultivated γ-proteobacterium, light-activated proton pumps have been widely detected among marine bacteria and, together with chlorophyll-based photosynthesis, are considered as an important axis responsible for primary production in the biosphere. Rhodopsins and related proteins show a high level of phylogenetic diversity; we focus on a specific class of bacterial rhodopsins containing the '3 omega motif.' This motif forms a stack of three non-consecutive aromatic amino acids that correlates with the B-C loop orientation and is shared among the phylogenetically close ion pumps such as the NDQ motif-containing sodium-pumping rhodopsin, the NTQ motif-containing chloride-pumping rhodopsin, and some proton-pumping rhodopsins including xanthorhodopsin. Here, we reviewed the recent research progress on these 'omega rhodopsins,' and speculated on their evolutionary origin of functional diversity.

Purification and Characterization of Manganese Superoxide Dismutase from Staphylococcus sciuri

  • Song, Chi-Hyun;Park, Eun-Kyung;Suh, Hyung-Joo;Lee, Yong-Se;Choi, Jang-Won;Ra, Kyung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.271-275
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    • 1999
  • The intracellular superoxide dismutase (SOD) from Staphylococcus sciuri was isolated to homogeneity by continuous steps, including ammonium sulfate fractionation, DEAE-ion-exchange chromatography, gel filtration, and phenyl hydrophobic gel chromatography. Pure SOD had a specific activity of 4,625 U/mg and was purified 158-fold with a yield of 31 % from a cell free extract. The molecular weight of the purified SOD was determined to be approximately 35.5 kDa by gel filtration and the enzyme was also shown to be composed of dimeric subunits on denaturing SDS-PAGE. The enzyme activity remained stable at pH 5 to 11 and also to heat treatment of up to $50^{\circ}C$ at pH 7.8, with 80% relative activity. The enzyme was insensitive to cyanide, hydrogen peroxide, and azide, indicating that it is a manganese-containing SOD. The EPR spectrum showed the enzyme containing manganese as a cofactor.

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Physicochemical and Biological Characteristics of Flavonoids Isolated from the Heartwoods of Rhus verniciflua (옻나무 목질부에서 분리된 플라보노이드의 이화학적 및 생물학적 특징)

  • Kwon, Sang-Hyuk;Kim, Gap-Tae;Lee, Kyung-Tae;Choi, Jung-Hye;Choi, Jong-Won;Park, Kun-Young;Park, Hee-Juhn
    • Korean Journal of Pharmacognosy
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    • v.31 no.3
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    • pp.345-350
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    • 2000
  • From the heartwood of Rhus verniciflua, four known flavonoids (1-4) were isolated along with an unknown one (5). Compounds 1-4 were identified to be garbanzol, sulfuretin, fisetin and fustin by NMR data. NMR data of 1-4 were fully assigned by the aids of 2D-NMR spectra. Among these compounds, only sulfuretin had significant cytotoxic and antioxidant ability at high concentrations. In addition, it seems likely that the 5-hydroxy-lacking flavonoids could not influence on the activity of laccase with cofactor of cupric cation, which catalyzes oxidative coupling reaction, in this plant.

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Protective Effects of Ulva lactuca Methanol Extracts against the Ultraviolet B-induced DNA Damage (자외선 B에 의해 유도되는 DNA 상해에 대한 참갈파래 메탄올 추출물의 보호 효과)

  • Jeong, Seula;Chung, Yuheon;Park, Jong Kun
    • The Korean Journal of Food And Nutrition
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    • v.33 no.3
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    • pp.309-316
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    • 2020
  • In this study, we investigated the protective effects of Ulva lactuca methanol extracts against ultraviolet B (UVB)-induced DNA damage in HaCaT cells. First, the contents of general and antioxidative nutrient contents of Ulva lactuca were measured. The moisture, carbohydrate, crude protein, crude fat and ash were 14.01%, 44.80%, 23.19%, 3.10% and 14.90%, respectively. Magnesium that acts as DNA repair enzyme cofactor was the most abundant mineral followed by Ca, P and Fe. The total phenolic and anthocyanoside contents of Ulva lactuca were 2.69 mg/g and 0.13 mg/g, respectively. Cells treated with Ulva lactuca methanol extracts for 24 hours post UVB exposure increased cell viability in a concentration-dependent manner compared to the non-treated control. Also, Ulva lactuca methanol extracts decreased the levels of UVB-induced DNA damage such as cyclobutane pyrimidine dimer and DNA damage response (DDR) proteins such as p-p53 and p21. These results suggest that Ulva lactuca methanol extracts comprising physiological active substances such as Mg, polyphenols and anthocyanosides promote DNA repair by regulating genes related with DDR.

Clinical Findings of 6-pyruvoyl-tetrahydropterins Synthase (PTPS) Deficiency in Korea (6-pyruvoyl-tetrahydropterins Synthase 결핍증의 임상적 고찰)

  • Yi, Youngsuk;Phil, Bae Seong;Lee, Jeong Ho;Lee, Dong Hwan
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.13 no.1
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    • pp.30-36
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    • 2013
  • 6-pyruvoyltetrahydropterin synthase (PTPS) deficiency is autosomal recessive disorder and the most common type of tetrahydrobiopterin (BH4) deficiency. It is caused by deficiency of PTPS, a cofactor involved in the biosynthesis of BH4 from guanosine triphosphate (GTP). Unlike classical phenylketonuria, which needs restriction of dietary phenylalanine for whole life, BH4 deficiency is treated by tetrahydrobiopterin, levodopa, and 5-hydroxytryptophan replacement. So it is important to make accurate diagnosis and initiate treatment as soon as possible for a better prognosis. There is no retrospective study of Korean patients undergoing long-term treatment for PTPS deficiency. We report 9 Korean patients with PTPS deficiency and their laboratory findings including BH4 loading tests, urine pterin tests, genotypes, dihydropteridine reductase (DHPR) activities and clinical manifestations including medication and developmental delay existence.

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Biotin Requirement for the Growth and Sporulation of Bacillus subtilis SNU816 in a Synthetic medium (Bacillus subtilis SNU816의 合成培地에서의 성장과 포자형성을 위한 Biotin 要求性에 관하여)

  • Lee, Oh-Hyoung
    • Korean Journal of Microbiology
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    • v.22 no.3
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    • pp.135-142
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    • 1984
  • The effect of biotin on the growth and sporulation of Bacillus subtilis SNU816 was investigated. When B. subtilis SNU816 was cultured on glucose as a sole carbon source, the growth was retarded markedly and usually ceased at early log phawe. But by addition of biotin to this medium, normal, rapid growth was restored. The growth rate was increased proportionally according to the concentration of exogenous biotin until it reached to 0.05㎍/ml, at which about three fold rapid growth was achieved. Also biotin was required for optimum sporulation for it facilitated the complete utilization of both glucose(Glc) and glutamic acid(Glu). Without biotin in Glc+Glu medium, about 40% of glutamic acid was remained unutilized. The dipicolinic acid content of cells cultured in Glc+Glu medium without biotin was markedly small and sporulation was suppressed before free spore release. Since biotin could be partiallyreplaced by one of TCA cycle intermediates such as oxalacetic acid, citric acid, or glutamic acid in enhancing growth in Glc medium, it was postulated that this strain might have a defect in converting pyruvate to oxalacetate which process is known to be mediated by pyruvate carboxylase that requires biotin as a cofactor.

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An FMN-containing NADH-quinone reductase from streptomyces sp (An FMN-Containing NADH-Quinone Reductase from Streptomyces sp.)

  • Youn, Hong-Duk;Lee, Jin-Won;Youn, Hwan;Lee, Jeong-Kug;Hah, Yung-Chil;Kang, Sa-Ouk
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.206-213
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    • 1996
  • NADH-quinone reductase was purified 22-fold from the cytosolic fraction of Streptomyces sp. Imsnu-1 to apparent hemogenity, with an overall yield of 9%, by the purification procedure consisting of ammonium, sulfate precipitation and DEAE Sephacryl S-200 and DEAE 5 PW chromatographies. Thes molecular mass of the enzyme determined by gel filtration chromatography was found to be 110 kDa. SDS-PAGE revealed that the enzyme consists of two sugunits with a molecular mass of 54 kDa. The enzyme contained 1 mol of FMN per subunit as a cofactor. The $A_{272}$ A$_{457}$ ratio was 6.14 and the molar extinction coefficients were calculated to be 20, 800 and 25, 400M$^{-1}$ $cm^{-1}$ / AT 349 AND 457 nm, respectively. The N-terminal sequence of the enzyme contained the highly conserved fingerprint of ADP-binding domain. The enzyme used NADH as an electron donor and various quinones as electron acceptors. Cytochrome c was practically inactive. Air-stable flavin semiquinone was produced by the addition of NADH to the enzyme. Also, naphthosemiquinone was detected in the reaction mixture containing the enzyme.

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Epoxide Hydrolase-catalyzed Hydrolytic Kinetic Resolution for the Production of Chiral Epoxides (에폭사이드 가수분해효소에 의한 동력학적 가수분해반응을 이용한 광학활성 에폭사이드 생산)

  • 이은열
    • KSBB Journal
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    • v.17 no.4
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    • pp.321-325
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    • 2002
  • Chiral epoxides are valuable intermediates for the asymmetric synthesis of enantiopure bioactive compounds. Microbial epoxide hydrolases (EHs) are newly discovered enzymes and versatile biocatalysts for the preparation of chiral epoxides by enantioselective hydrolysis of cheap and easily available racemic epoxide substrates. EHs are commercially potential biocatalysts due to their characteristics such as high enantioselectivity, cofactor-independent catalysis, and easy-to-Prepare catalysts. In this Paper, recent progresses in biochemistry and molecular biology of EH and developments of novel reaction systems are reviewed to evaluate the commercial feasibility of EH-catalyzed hydrolytic kinetic resolution for the production of chiral epoxides.

Microbial styrene monooxygenase-catalyzed asymmetric synthesis of enantiopure styrene oxide derivatives (미생물 유래 Styrene monooxygenase를 이용한 광학활성 styrene oxide 유도체의 비대칭합성)

  • Lee, Eun-Yeol;Park, Sung-Hoon
    • KSBB Journal
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    • v.24 no.3
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    • pp.239-245
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    • 2009
  • Enantiopure styrene oxide derivatives are versatile building blocks for the synthesis of enantiopure pharmaceuticals. Styrene monooxygenase (SMO) catalyzes an asymmetric addition of an oxygen atom into a double bond of vinylaromatic compounds. SMO is a commercially potential biocatalyst to synthesize a variety of enantiopure epoxides with high enantiopurity and recovery yield. In this paper development of SMO biocatalyst and commercial feasibility of SMO-catalyzed asymmetric synthesis of enantiopure stylers oxide derivatives are reviewed.

Effect of TGF-${\beta}1$ on Osteoclast Differentiation

  • Park, Su-Jin;Ko, Jea-Seung;Kim, Hyun-Man
    • International Journal of Oral Biology
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    • v.30 no.4
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    • pp.135-141
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    • 2005
  • Although it has been known that TGF-${\beta}1$ acts as a crucial cofactor in osteoclast differentiation, its mode of action is still unclear. In the present study, we studied the effect of TGF-${\beta}1$ on the differentiation of osteoclast depending on the developmental stages. Murine bone marrow cells were induced to differentiate into mature osteoclasts in the presence of receptor activator of NF-${\kappa}B$ ligand (RANKL) and macrophage colony stimulating factor (M-CSF). In the early stage of the differentiation TRAP(-) mononuclear precursor cells were obtained from nonadherent M-CSF dependent bone marrow cells, which further differentiated into mature osteoclasts. TGF-${\beta}1$ stimulated osteoclast differentiation, which was stronger when cells were stimulated by TGF-${\beta}1$ in the early stage than the later differentiation. TGF-${\beta}1$ increased the expression of RANK and synergistically stimulated RANKL-induced activation of NF-${\kappa}B$ MAP kinase in TRAP(-) mononuclear precursor cells. These results suggest that activation of osteoclast differentiation by TGF-${\beta}1$ may be ascribed to the both increased expression and activation of RANK in the osteoclast differentiation, especially in the early stage of differentiation.