• Title/Summary/Keyword: Carbon Source

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Characterization and N Terminal Amino Acid Sequence Analysis of Catechol 1,2-Dioxy-genase from Benzoate Degrading Acinetobacter sp. KS-1 (Benzoate 분해세균 Acinetobacter sp. kS-1에서 분리된 catechol 1,2-dioxygenase의 특성 및 N 말단 아미노산 서열 분석)

  • 오계헌;송승열;김승일;윤경하
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.74-80
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    • 2002
  • The purpose of this work was to investigate the characterization and sequence of catechol 1,2-dioxygenase (Cl,2O) purified from Acinetobacter sp. KS-1 which was grown on benzoate as a sole carbon source. Cl,2O demonstrated its enzyme activity to catechol and 4-methylcatechol. The optimum temperature of Cl,2O was $35^{\circ}C$, and the optimal pH was in the range from pH 7.5 to 9.0. $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of Cl,2O. Molecular weight of the enzyme was determined to approximately 36 kDa by SDS-PAGE and 7-terminal amino acid sequence of Cl,2O was analyzed as $^{1}MNYQQIDALVKQMNVDTAKG^{20}$and exhibited 95% sequence homology with that of Cl,2O from Acinetobacter radioresistens In addition, trypsin digestion and peptide mapping were performed for internal sequencing analysis. Molecular weights of three digested peptide fragments were analyzed as 966.3 Da, 1933.8 Da and 2081.7 Da by MALDI-TOF, which were matched with each internal sequences $^{1}SQSDFNLRR^{9}\, ^{1}HGNRPSHVHYFNSAPGYR^{18}\, ^{1}TIEGPLYVAGAPESVGFAR^{19}$) of. A. radioresistens. PCR product was amplified with the degenerated primers derived from N-terminal and each internal amino acid sequences.

Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1 (Methylophaga aminosulfidovorans SKI bfmo 오페론의 클로닝 및 구조 분석)

  • Lim Hyun Sook;Goo Jae Whan;Kim Lee Hyun;Kim Si Wouk;Cho Eun Hee
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.1-7
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    • 2005
  • Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP) can utilize trimethylamine as a sole carbon, nitrogen, and energy source. The bacterial flavin-containing monooxygenase (bFMO) gene was identified in the strain and the recombinant enzyme expressed in E. coli oxidized trimethylamine. To study the function and regulation of the bfmo, over 8,000 nucleotide sequences of the neighboring regions including the bfmo were determined. Three open reading frames proceeding to the bfmo gene encoded analogues to highly conserved nitrate/nitrite sensing two-component system regulators and a methyl accepting protein. Two small open reading frames just downstream of the bfmo gene showed no similar proteins of known functions but the sequences were conserved among other bacteria. Reverse transcription-polymerase chain reaction analysis showed that the six putative genes consisted of three transcription units. The three regulatory genes located upstream of the bfmo gene formed two separate transcription units. The bfmo and the two downstream genes were transcribed from a single promoter.

Production of Rubber-Elastic Polyhydroxyalkanoates by Pseudomonas sp. HJ-2 (Pseudomonas sp. HJ-2를 이용한 고무탄성 Polyhydroxyalkanoate의 생산)

  • 정정욱;최강욱;김영백;이영하
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.155-160
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    • 2000
  • Pseudomnas sp. HJ-2 is capable of producing a rubber-elastic polyhydroxyalkanoate (PHA) consisting of 3- hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyheptanoate (3HHp) from heptanoic acid as the sole carbon source. The polyester produced was a blend of poly(3HB-co-3HV) and poly(3HHp). Although the mixing of poly(3HHp) fraction to poly(3HB-co-3HV) resulted in a decrease of modulus, the sole fraction of poly(3HB-co-3HV) with a high molar fraction of 3HV was shown to be an elastomer with the maximum percent strain of 740%. The biomass yield and the PHA synthesis were relatively high when the initial heptanoic acid concentration was 40 mM, and were significantly decreased when the substrate concentration exceeded 50 mM. The accumulation of PHA was stimulated by deficiency of nitrogen and phosphorus in the medium. The PHA contents and its monomeric compositions were greatly affected by pH and oxygen transfer rate. At pH 7.5, poly(3HB-~0.38% 3HV) was produced from heptanoic acid and a mixture of 95% 3HHp and 5% 3HV was produced at pH 8.0. Increased conten1 of 3HHp in the polyesters with lhe increasing oxygen transfer rate by agitation speed a1 a fixed aeration rate was observed.

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Production of Biosurfactant by Pseudomonas sp. SW1 for Microbial Remediation of Oil Pollution (유류오염방제를 위한 Pseudomonas sp. SW1로부터 생물계면활성제의 생산)

  • Son, Hong-Joo;Suk, Wan-Su;Lee, Geon;Lee, Sang-Joon
    • Korean Journal of Microbiology
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    • v.33 no.3
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    • pp.193-198
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    • 1997
  • Microorganisms capable of producing biosurfactant were isolated from oil-contaminated soils and seawater. Among them, the selected strain SW1 was identified as Pseudomonas sp. by taxonomical characteristic tests, and so tentatively named Pseudomonas sp. SWI. The optimal temperature and initial pH for biosurfactant production were TEX>30^{\circ}C.$ and 7.0, respectively. The optimal medium composilion for the production of biosurfactant by Pseudomonas sp. SW1 were hexadecane of 2.0%, yeast extract of 0.04%, $K_{2}HPO_4$ of 0.02%, $KH_2PO_4$ of 0.03% and $MgSO_4$ center dot $7H_2O$ of 0.04%, respectively. Under the above conditions, minimum wrface tension was 32 mN/m after incubation of 2 days. The biosurfactant was produced during initial stationary phase in the optimal medium. Pseudotnonas sp. SWl utilized various hydrocarbons such as Bunker oils, n-alkanes and branched alkanes as a sole carbon source.

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Simultaneous Removal of Phenol and Hexavalent Chromium by Rhodococcus sp. CP01 (Rhodococcus sp. CP01에 의한 페놀과 6가 크롬이온의 동시 제거)

  • 최광현;오영숙;김병동;최성찬
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.279-284
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    • 2000
  • Simultaneous reduction of Cr(VI) and degradation of phenol was observed in batch and bench-scale continuous stirred tank reactors using Rhodococcus sp. CP01 isolated from leachate. The strain CP01, which was capable of utilizing phenol as a sole source of carbon and energy, completely reduced added hexavalent chromium (0.25 mM) to its trivalent form during 60 hr batch assay under optimal conditions (pH 7.0 and 1,000 mg/L of phenol concentration). The rates of Cr(VI) reduction and phenol degradation were estimated as 4.17 $\mu$M Cr(VI) and 38.4 mg phenol.$L^{-1}{\cdot}hr^{-1}$, respectively. The continuous culture experiment was conducted for 46 days using synthetic feed containing different levels of chromate (0.0625 to 0.25 mM) and phenol(1,000 to 4,000 mg/L). With a hydraulic retention time of 100 hr, Cr(VI) reduction efficiency was mostly 100% for influent Cr(VI) and phenol concentrations of 0.125 mM and 3,000 mg/L, respectively. During quasi-steady-state operation, specific rate of Cr(VI) reduction was calculated as 0.34 mg Cr(VI).g $protein^{-1}{\cdot}hr^{-1}$ which was comparable to reported values obtained by using glucose as growth substrate. The results suggest the potential application of biological treatment for detoxification of wastewater contaminated simultaneously with Cr(VI) and pheonol.

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Geochemical Characteristics of the Continental Shelf and Slope Sediments off the Southeastern Coast of Korea (한국 동남해역 대륙붕과 대륙사면 표층퇴적물의 지화학적 특성)

  • Lee, Chang-Bok;Park, Yong-Ahn;Kang, Hyo-Jin;Kim, Dae-Chul
    • The Korean Journal of Quaternary Research
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    • v.5 no.1
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    • pp.15-31
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    • 1991
  • A total of 90 surface sediment samples, collected from the continental margin area bordering east and southeast coast of Korea, were subject to the geochemical analyses with the aim of filling the gap in our knowledge of this environment. The analyzed items included the major elements (Al, Fe, Mg, Ca, K, Na, p and Mn), organic carbon, and some trace metals (Ba, Co, Cu, Sr and Zn). The sediment grain-size exerted a predominant influence on the contents of most elements, with the exceptions of Ca, K, Sr and Ba. The Ca and Sr contents, being closely interrelated each other, were mainly controlled by the calcium carbonate content. The K content, on the other hand, appeared to be influenced by both illite and feldspar. The Ba content showed a certain relationship with that of K, suggesting a common source of these two elements; potassium feldspar. The R-mode factor analysis result also reaffirmed the above-mentioned controlling factors on the sediment geochemistry. The grain-size dependency of trace metals obscures their areal distribution pattern from the total contents. However, with the metal/aluminum ratios we could differentiate the subtle difference in the metal enrichment. Hence, sediments of the southern coastal area appear to receive some anthropogenic inputs of metals, though the effect is still negligible.

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Production of $\beta$-Mannanase and $\beta$-Mannosidase from Sporolactobacillus sp. M201. (Sporolactobacillus sp. M201 균주에 의한 $\beta$-Mannanase와 $\beta$-Mannosidase의 생산)

  • 박원식;김화영;최용진
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.232-237
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    • 1998
  • A bacterial strain producing high levels of an extracellular ${eta}$-mannanase and intracellular ${eta}$-mannosidase and ${alpha}$-galactosidase was isolated from soil. The strain isolated was identified as a strain of Sporolactobacillus sp. and designated as Sporolactobacillus sp. M20l. Synthesis of ${eta}$-mannanase by Sporolactobacillus sp. M20l was induced by sucrose, maltose, or locust bean gum. The highest induction rate was obtained with 2% locust bean gum added to the culture medium as a sole carbon source. On the other hand, induction of ${eta}$-mannosidase was observed only with locust bean gum. The optimal media for the enzyme production were established as follows: for ${eta}$-mannanase; 2% locust bean gum, 0.5% peptone, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 6.0), and for ${eta}$-mannosidase; 2% locust bean gum, 0.5% yeast extract, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 5.0). The optimal culture temperatures for production of ${eta}$-mannanase and ${eta}$-mannosidase were found to be 37$^{\circ}C$ and 3$0^{\circ}C$, respectively. Under the optimal culture conditions, the production of ${eta}$-mannanase and ${eta}$-mannosidase reached the highest levels of 10.6 units/ml and 1.35 units/ml after 30 h and 24 h cultivation, respectively.

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Comparative Analysis of Aniline Dioxygenase Genes from Aniline Degrading Bacteria, Burkholderia sp. HY1 and Delftia sp. HY99. (Aniline 분해균주 Burkholderia sp. HY1과 Delftia sp. HY99에서 유래된 Aniline Dioxygenases 유전자의 비교 분석)

  • Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.104-111
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    • 2007
  • In this study, aniline dioxygenase genes responsible for initial catabolism of aniline in Burkholderia sp. HY1 and Delftia sp. HY99 were cloned and the amino acid sequences were comparatively analyzed, which already have been reported as bacteria utilizing aniline as a sole source of carbon and nitrogen, B. sp. HY1 was found to have at least a plasmid, and the plasmld-cured strain, B. sp. HY1-PC obtained using mitomycin C was tested with wild type strain to investigate whether the former maintained the degradability for aniline. This proved that the aniline oxygenase gene from B. sp. HY1 was located in chromosomal DNA, not in plasmid DNA. Aniline dioxygenase small subunits from B. sp. HY1 and D. sp. HY99 were found, based on 146 amino acids, to share 79% similarity. Notably, ado2 genes from B. sp. HY1 and D. sp. HY99 which were found to be terminal dioxygenase of aniline dioxygenase small subunit showed 99% similarity in the deduced amino acid sequences with tdnA2 of Frateuria sp. ANA-18 and danA2 of D. sp. AN3, respectively. Besides, enzyme assay and amino acid sequence analysis of catechol dioxygenase supported the previous report that B. sp. HY1 might occupy ortho-cleavage pathway using catechol 1,2-dioxygenase, while D. sp. HY99 might occupy catechol 2,3-dioxygenase for meta-cleavage pathway.

Isolation and Characterization of a Diesel-Degrading Bacterium, Gordonia sp. SD8 (디젤 분해 세균 Gordonia sp. SD8 분리 및 특성)

  • Hong, Sun-Hwa;Kim, Ji-Young;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.335-339
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    • 2010
  • A diesel-degrading bacterium, Gordonia sp. SD8, was isolated from soil contaminated with petroleum, and its diesel degradation was characterized in a soil as well as a liquid culture system. SD8 could grow in the mineral salt medium supplemented with diesel as a sole carbon and energy source. The maximum specific growth rate ($0.67{\pm}0.05\;d^{-1}$) and diesel degradation rate ($1,727{\pm}145$ mg-TPH $L^{-1}\;d^{-1}$) of SD8 showed at 20,000 mg-TPH $L^{-1}$ and $30^{\circ}C$, and then this bacterium could degrade high strength of diesel of 40,000 mg-TPH $L^{-1}$. The residual diesel concentration in the inoculated soil with SD8 was 3,724 mg-TPH kg-dry $soil^{-1}$ after 17 days, whereas the diesel concentration in the non-inoculated soil was $8,150{\pm}755$ mg-TPH kg-dry $soil^{-1}$. These results indicate that Gordonia sp. SD8 can serve as a promising microbial resource for the bioremediaion of contaminated soil with petroleum hydrocarbons including diesel.

Characterization of Starch-Utilizing Yeast Saccharomycopsis fibuligera Isolated from Nuruk (누룩으로부터 분리된 전분대사 효모 Saccharomycopsis fibuligera 균주의 생육특성)

  • Choi, Da-Hye;Park, Eun-Hee;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.407-412
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    • 2014
  • A number of Saccharomycopsis fibuligera strains that can hydrolyse and utilize starch as a carbon source were isolated from nuruk, a traditional Korean starter for rice wine fermentation, and their specific growth rates on starch-containing medium were compared to choose the prominent strain. S. fibuligera strain MBY1320 showed a higher growth rate at $42^{\circ}C$ than that of strain S. fibuligera KCTC7806, indicating that S. fibuligera MBY1320 has more thermo-tolerant machinery for starch hydrolysis and utilization than KCTC7806. Although the activity of ${\alpha}$-amylase at $30^{\circ}C$ was significantly lower for S. fibuligera MBY1320 than KCTC7806 (3,812.5 U vs. 14,878.5 U), S. fibuligera MBY1320 showed a much higher glucoamylase activity at $42^{\circ}C$ than S. fibuligera KCTC7806 (5,048.9 U vs. 13,152.3 U). Thus, a new S. fibuligera strain, with a higher starch-hydrolysing activity at elevated temperatures than that of other types of strain, this study reports.