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Cytochrome P450 monooxygenase analysis in free-living and symbiotic microalgae Coccomyxa sp. C-169 and Chlorella sp. NC64A

  • Mthakathi, Ntsane Trevor;Kgosiemang, Ipeleng Kopano Rosinah;Chen, Wanping;Mohlatsane, Molikeng Eric;Mojahi, Thebeyapelo Jacob;Yu, Jae-Hyuk;Mashele, Samson Sitheni;Syed, Khajamohiddin
    • ALGAE
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    • v.30 no.3
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    • pp.233-239
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    • 2015
  • Microalgae research is gaining momentum because of their potential biotechnological applications, including the generation of biofuels. Genome sequencing analysis of two model microalgal species, polar free-living Coccomyxa sp. C-169 and symbiotic Chlorella sp. NC64A, revealed insights into the factors responsible for their lifestyle and unravelled biotechnologically valuable proteins. However, genome sequence analysis under-explored cytochrome P450 monooxygenases (P450s), heme-thiolate proteins ubiquitously present in species belonging to different biological kingdoms. In this study we performed genome data-mining, annotation and comparative analysis of P450s in these two model algal species. Sixty-nine P450s were found in two algal species. Coccomyxa sp. showed 40 P450s and Chlorella sp. showed 29 P450s in their genome. Sixty-eight P450s (>100 amino acid in length) were grouped into 32 P450 families and 46 P450 subfamilies. Among the P450 families, 27 P450 families were novel and not found in other biological kingdoms. The new P450 families are CYP745-CYP747, CYP845-CYP863, and CYP904-CYP908. Five P450 families, CYP51, CYP97, CYP710, CYP745, and CYP746, were commonly found between two algal species and 16 and 11 P450 families were unique to Coccomyxa sp. and Chlorella sp. Synteny analysis and gene-structure analysis revealed P450 duplications in both species. Functional analysis based on homolog P450s suggested that CYP51 and CYP710 family members are involved in membrane ergosterol biosynthesis. CYP55 and CYP97 family members are involved in nitric oxide reduction and biosynthesis of carotenoids. This is the first report on comparative analysis of P450s in the microalgal species Coccomyxa sp. C-169 and Chlorella sp. NC64A.

Isolation of Marine Bacteria Killing Red Tide Microalgae II. Isolation and Algicidal Properties of Pseudomonas sp. LG-2 Possessing Killing Activity for Dinoflagellate, Prorocentrum micans (적조생물 살조세균 탐색 II. 적조생물 Prorocentrum micans 살조세균 Pseudomonas sp. LG-2의 분리와 살조특성)

  • LEE Won-Jae;PARK Young-Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.6
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    • pp.852-858
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    • 1998
  • We have isolated a bacterial strain that tends to kill P. micans from the mixed culture of p. minns plus seawater filtrate (poresize, 0.8 $\mu$m) collected at Masan bay in July 1996, in which the mixed culture grown in the f/2 medium. According to the experimental results of the isolated bacterium such as fatty acids analysis, morphological and biochemical characteristic tests, the strain was supposed to be a Pseudomonas and then it was named as Pseudomonas sp. LG-2. The killing effect of Pseudomonas sp. LG-2 against P. micans was proportionally increased with the concentrations of culture filtrate (pore size, 0.8 $\mu$m) is well as with the number of bacterium inoculated. In the mixed culture inoculated with $1.3\times10^6$ cells/ml of Pseudomonas sp. LG-2, the number of P. micans (2,000 cells/ml) was gradually decreased and then killed below 100 cells/ml within 7 days. In addition, the culture filtrate with $30\%$ of final concentration revealed a significant killing effect against P. micans around 3 days after culture. In the relationship between killing effects and growth stage of Pseudomonas sp. LG-2, the culture filtrate at lag phase has little effects on P. micans. In constant, the culture filtrate at mid-log phase showed the killing effect by decreasing P. micans to 112 in number within 5 days. In particular, the culture filtrate at stationary phase showed a significant killing effect against P. micans in which the majority of it was killed after 3 day culture. The species specificity of killing effects of Pseudomonas sp. LG-2 against 5 species of dinoflagellate was only found in P. micans and Scrippsiella trochoidea.

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Descriptions of Four New Species of Predatory Nematodes (Mononchida) From Korea (韓國産 捕食線蟲(Mononchida: Nematoda)의 4 新種 기재)

  • Choi, Young-Eoun;Khan, Zakaullah;Lee, Sung-Min
    • The Korean Journal of Soil Zoology
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    • v.4 no.2
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    • pp.89-100
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    • 1999
  • Four new and a known spacies of the order Mononchida were described and illustrated. Iotonchus obtusus sp. n. was 2.8 mm long, a=33, b=4.2, c=61, V=68%, buccal cavity=61${\times}$45 mm, and is characterized by having basally situated dorsal tooth, presence of vulval papillae and in having short, hemispherical tail with thick cuticle at terminus. Miconchus vulvapapillatum sp. . was 2.7-3.6 mm long, a=29-36, b=4.1-4.5, c=18.4-21, V=65-69%, buccal cavity=53-61${\times}$29-33 mm, spicules=132-137 mm, ventromedian supplements 28-31, and was characterized by having 5-8 pre- and post vulval papillae in contiguous series, and three pairs of vulval glands. Clarkus koreanus sp. n. was 1.1-1.3 mm long, a=27.5-28.8, b=3.5-3.9, c=12-14.5, V=60-64%, buccal cavity=24-28${\times}$13.5-15 mm, and was characterized by well offset lip region, amphids situated well below to dorsal tooth apex, and vulva elevated, with vulval flap. Coomansus ulsani sp. n. was 1.2-1.5 mm long, a=23.5-26, b=3.4-3.8, c=13.6-14.8, V=65-68%, buccal cavity=36-39${\times}$21-23 mm and was characterized by well offset lip region and a thin longitudinal ridge on vertical walls of stoma.

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Physicochemical and Dyeing Properties of Microbial Prodiginine from Zooshikella sp. (미생물 Prodiginine 색소의 물리화학적 특성 및 섬유염색성)

  • Kim, Yong-Sook;Choi, Jong-Myoung
    • Journal of the Korean Society of Clothing and Textiles
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    • v.35 no.4
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    • pp.431-441
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    • 2011
  • Microbial colorants produced from Zooshikella sp. were developed as a reddish dye for fabrics. The reddish colorants were extracted from cell mass of Zooshikella sp. using 100% ethanol and were identified as prodiginine by 1H-NMR and FT-IR analysis. Microbial prodiginine had a maximum spectrophotomatric absorbance at 530nm and were chemically stable and 30 to $60^{\circ}C$. The microbial prodiginine could dye natural fibers such as cotton, silk, and wool as well as synthetic fibers such as nylon. The maximum K/S values of the dyed fiber were shown at 540 run with a color appearance of RP (reddish purple). Silk and nylon had an excellent dyeability among the experimental fibers. The optimum pH for the dyeing of experimental fibers was at pH 3.0 and dyeability was improved as the temperature increased. The cover change of dyed multifiber fabrics with the microbial prodiginine were measured after washing with detergents and a dry cleaning solvent for the selection of a proper fabric against microbial prodiginine. Among the experimental fibers, silk and nylon did not show significant color change after washing. Therefore, under the criteria of dyeability, silk and nylon were excellent fabrics for being dyed by microbial prodiginine.

Ecological Distribution of Endomycorrhizal Fungi in Pogil-do in Tadohae-haesang National Park

  • Seo, Hyun-Chang;Kim, Chang-Jin;Kim, Shin-Duk
    • Journal of Microbiology and Biotechnology
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    • v.2 no.1
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    • pp.66-71
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    • 1992
  • The ecological distribution of endomycorrhizas in evergreen woody species native to the evergreen forest ecosystem of Tadohae-haesang National Park in southern Korea in February, 1989 was studied. The abundance and diversity of vesicular-arbuscular (VA) mycorrhizal fungi were also determined. The spore densities ranged from 14 to 326 per 100 g of soil. Most of the spores of mycorrhizal fungi collected from 25 soil samples belonged to the genera Glomus and Gigaspora. The frequency and number of spores in Camellia japonica varied with location. Spores belonging to the genus Gigaspora were not found in Camellia japonica in Yesongri evergreen forests adjacent to the sea. Glomus sp. was the major constituent of the spore assemblage at this site. The most abundant species in Camellia japonica in the Yesongri evergreen forests in Pogildo was Glomus borealis. In the soil of a mountain at Buwhangri, in the central location of the island at an elevation of 250 m, Gigaspora sp. was present and Glomus sp. was a major constituent of the spore assemblage. In the urban area of Haenam spore densities were much higher than in the Pogildo area. The most abundant species in Camellia japonica in the urban area of Haenam was Gigaspora sp..

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Purification and Characterization of the Intracellular Alginase from Vibrio sp. AL-145 (알긴산 분해균 Vibrio sp. AL-145가 생산하는 균체내 효소의 정제 및 특성)

  • Joo, Dong-Sik;Lee, Jung-Suk;Park, Jung-Je;Cho, Soon-Yeong;Ahn, Chang-Bum;Lee, Eung-Ho
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.432-438
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    • 1995
  • The intracellular alginase from Vibrio sp. AL-145 was purified by ion chromatography on DEAE-Cellulose column, Q-Sepharose column, and gel filtration on Sephadex G-100 column. The optimum pH and temperature for the activity of the purified intracellular enzyme were 8.0 and 37$\circ$C, respectively. The enzyme was stable at the pH range of 7.5-8.5, and at 30$\circ$C for 30 min. The molecular weight of the intracellular enzyme was estimated to be about 23, 000 daltons by SDS-polyacrylamide gel electrophoresis. NaCl was required for enzyme activity and the optimum concentration was 0.5 M. The activity of intracellular enzyme was inhibited by Co$^{2+}$, Hg$^{2+}$, Zn$^{2+}$, 0-phenanthroline, $\rho$-CMB, EDTA and iodoacetate, and stimulated by Ca$^{2+}$, L-cysteine and 2-mercaptoethanol. This enzyme was an alginase specifically degrading alginic acid.

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A New IgG Immunoblot Kit for Diagnosis of Toxoplasmosis in Pregnant Women

  • Khammari, Imen;Saghrouni, Fatma;Lakhal, Sami;Bouratbine, Aida;Said, Moncef Ben;Boukadida, Jalel
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.493-499
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    • 2014
  • The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of Toxoplasma gondii tachyzoites and assessed it by testing 154 positive, 100 negative, and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21, and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as the positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II $IgG^{(R)}$ kit recently recommended as a confirmatory test (96.7% of concordance).

Purification and characterization of an alkaline protease produced by a xanthomonas sp. YL-37

  • Lee, Chang-Ho;Kim, Hee-Sik;Seok, Kwon-Gi;Oh, Hee-Mock;kang sang mo;Kwon, Tae-Jong;Yoon, Byung-Dae
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.115-119
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    • 1995
  • The alkaline protease of Xanthomonas sp. YL-37 has been purified, and the properties of the enzyme investigated. The alkaline protease of Xanthomonas sp. YL-37 was purified form crude enzyme by ammonium sulfate fractionation, CM-cellulose ion exchange chromatography, and Sephadex G-100 gel filtration. Through the series of chromatographies, the enzyme was purified to homogenecity with specific activity of 4.23 fold higher than that of the crude broth. The molecular weight of the purified protease has been estimated to be 62 KDa on SDS-polyacrylamide gel electrophoresis. The optimal pH and temperature for alkaline protease activity were 11.0 and 50.deg.C, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to 50.deg.C. Enzyme activity was lost up to 50% on heating at 70.deg.C for 30 minutes. The activity of alkaline protease was inhibited by Cu$\^$2+/, Zn$\^$2+/, Hg$\^$2+/, PMSF, and activated by Mn$\^$2+/ and Ca$\^$2+/. The $K_{m}$ value for casein as a substrate was 4.0 mg/ml.

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Cloning and Molecular Characterization of groESL Heat-Shock Operon in Methylotrophic Bacterium Methylovorus Sp. Strain SS1 DSM 11726

  • Eom, Chi-Yong;Kim, Eung-Bin;Ro, Young-Tae;Kim, Si-Wouk;Kim, Young-Min
    • BMB Reports
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    • v.38 no.6
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    • pp.695-702
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    • 2005
  • The groESL bicistronic operon of a restricted facultative methylotrophic bacterium Methylovorus sp. strain SS1 DSM 11726 was cloned and characterized. It was found to consist of two ORFs encoding proteins with molecular masses of 11,395 and 57,396 daltons, which showed a high degree of homology to other bacterial GroES and GroEL proteins. The genes were clustered in the transcription order groES-groEL. Northern blot analyses suggested that the groESL operon is transcribed as a bicistronic 2.2-kb mRNA, the steady-state level of which was markedly increased by temperature elevation. Primer extension analysis demonstrated one potential transcription start site preceding the groESL operon, which is located 100bp upstream of the groES start codon. The transcription start site was preceded by a putative promoter region highly homologous to the consensus sequences of Escherichia coli ${\sigma}^{32}$-type heat shock promoter, which functioned under both normal and heat shock conditions in E. coli. Heat shock mRNA was maximally produced by Methylovorus sp. strain SS1 approximately 10min after increasing the temperature from 30 to $42^{\circ}C$. The groESL operon was also induced by hydrogen peroxide or salt shock.

Styrene Degradation in a Polyurethane Biofilter Inoculated with Pseudomonas sp. IS-3

  • KIM JAISOO;RYU HEE WOOK;JUNG DONG JIN;LEE TAE HO;CHO KYUNG-SUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1207-1213
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    • 2005
  • In a search for bacteria capable of degrading styrene better than previously isolated strains, bacterium IS-3 was isolated from activated sludge and found to be most closely related to Pseudomonas sp. Styrene degradation by this strain was tested in liquid cultures and polyurethane-packed biofilters. In liquid cultures, the rate of styrene degradation by this bacterium increased from 24.93 to $76.53\;{\mu}mol\;g^{-1}\;DCW\;H^{-1}$ for an initial mass range from 8.7 to $34.8{\mu}mol$. The maximum styrene elimination capacity was 580-635 $g/m^{3}\cdot$h at a space velocity (SV) of 50-200/h. The critical elimination capacities guaranteeing $95\%$ removal of the input styrene were determined to be 635, 170, and 38 $g/m^{3}\cdot$h, respectively, at SVs of 50, 100, and 200/h. Kinetic analysis revealed that the maximum styrene elimination velocity ($V_{m}$) for this biofilter was 1,000 g/m$\cdot$h, and the saturation constant ($K_{m}$) was 454 ppmv. Together, these results suggest that a polyurethane biofilter containing Pseudomonas sp. IS-3 could have potential practical applications for the effective removal of styrene gas.