• 제목/요약/키워드: Pseudomonas sp

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Pseudomonas sp.로부터 알칼리내성 amylase의 정제 및 특성 확인 (Purification and Characterization of Alkali-resistant Amylases from Pseudomonas sp.)

  • 이정은;전덕영
    • 한국식품과학회지
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    • 제40권1호
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    • pp.70-75
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    • 2008
  • 두 개의 amylase를 호알칼리성 Pseudomonas sp. KFCC 10818의 배양액으로부터 정제하여 그 특성을 조사하였다. 정제된 효소의 분자량은 각각 50 kDa과 75 kDa이었다. 이 효소들의 최적반응온도는 각각 $35^{\circ}C$$40^{\circ}C$였으며 50 kDa의 효소는 칼슘이온에 의하여 효소활성이 두 배로 촉진되었다. 이 두 효소는 최적 pH가 6-8 부근이었으며 pH 12의 조건에서도 효소활성을 유지하는 알칼리내성을 나타냈다. Maltooligosaccharide이나 soluble starch로부터 maltose와 maltotriose를 최종 효소반응산물로 생산하였다. 두 amylase는 N-말단 아미노산 서열이 각각 QTVPKTTFV와 DTVPGNAFQ로 분석되었다.

Pseudomonas sp. DJ77 균주에서 Extradiol Dioxygenase를 암호화하는 phnQ 유전자의 클로닝과 대장균에서의 발현 (Cloning of phnQ Gene Encoding Extradiol Dioxygenase from Pseudomonas sp. DJ77 and Its Expression in Escherichia coli)

  • 신희정;박용춘;민경희;김치경;임재윤;김영창
    • 미생물학회지
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    • 제33권1호
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    • pp.22-26
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    • 1997
  • Pseudomonas sp. DJ77의 게놈 library로부터 phenanthrene 분해에 관련된 유전자를 포함하는 약 5-kb의 XhoI 절편을 pBLUESCRIPT SK(+)로 클로닝하였으며, 이 재조합 plasmid를 pUPX5라 명명하였다. 이 재조합 균주에 catechol과 2,3-dihydroxybiphenyl 용액을 분무하면 노란색의 meta-cleavage 화합물이 생성됨을 관찰 할 수 있었다. 그리고 효소 활성을 측정한 결과 catechol에서보다 2,3-dihydroxybiphenyl에 더 큰 활성을 나타냈다. 이 부분에 존재하는 2,3-dihydroxybiphenyl 1,2-dioxygenase 유전자의 위치를 결정하고, 이 유전자를 phnQ라 명명하였다.

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Pseudomonas sp. EL-G527에 의한 환경친화성 생물계면활성제의 생산최적조건 (Optimal Culture Conditions for Production of Environment-Friendly Biosurfactant by Pseudomonas sp. EL-G527)

  • 차미선;임은경;이근희;조순자;손홍주;이상준
    • 한국환경과학회지
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    • 제11권3호
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    • pp.177-182
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    • 2002
  • A biosurfactant-producing microorganism was isolated from activated sludge by enrichment culture when grown on a minimal salt medium containing n-hexadecane as a sole carbon source. This microorganism was identified as Pseudomonas sp. and it was named Pseudomonas sp. EL-G527. It's optimal culture condition is 2% n-hexadecane, 0.2% NH$_4$NO$_3$, 0.3% KH$_2$PO$_4$, 0.3% $K_2$HPO$_4$, 0.02% MgSO$_4$ㆍ7$H_2O$, 0.0025% CaCk$_2$ㆍ6$H_2O$, 0.0015% FeSO$_4$ㆍ7$H_2O$ in 1$\ell$ distilled water and initial pH 7.0. Cultivation was initiated with a 2% inoculum obtained from starter cultures grown in 30 $m\ell$ of the same medium in 250 $m\ell$ flask. They were cultivated at 3$0^{\circ}C$ in reciprocal shaking incubator and the highest biosurfactant production was observed after 4 days.

Cloning and Sequence Analysis of the xyIL Gene Responsible for 4CBA-Dihydrodiol Dehydrogenase from Pseudomonas sp. S-47

  • 박동우;이상만;가종옥;김지경
    • 미생물학회지
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    • 제38권4호
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    • pp.275-275
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    • 2002
  • Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as carbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.

Antibacterial Activity of an Ethyl Acetate Extract of Pseudomonas sp. UJ-6 against Methicillin-Resistant Staphylococcus aureus

  • Lee, Dae-Sung;Eom, Sung-Hwan;Je, Jae-Young;Kim, Young-Mog;Lee, Myung-Suk;Kim, Young-Man
    • Fisheries and Aquatic Sciences
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    • 제16권2호
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    • pp.79-84
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    • 2013
  • In an effort to discover an alternative antibiotic for treating infections with methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas sp. UJ-6, a marine bacterium that exhibited antibacterial activity against MRSA, was isolated. The culture broth and its ethyl acetate extract exhibited bactericidal activity against MRSA. The extract also exhibited antibacterial activity against gram-negative bacteria, which were not susceptible to vancomycin. The treatment of MRSA with the extract resulted in abnormal cell lysis. The extract retained >95% of its anti-MRSA activity after heat treatment for 15 min at $121^{\circ}C$. Thus, although most antibiotics are unstable under conditions of thermal stress, Pseudomonas sp. UJ-6 produces a heat-stable anti-MRSA substance. The results of this study strongly suggest that Pseudomonas sp. UJ-6 can be used to develop a novel, heat-stable, broad-spectrum antibiotic for the treatment of MRSA infections.

Mercury Resistance and Removal Mechanisms of Pseudomonas sp. Isolated Mercury-contaminated Site in Taiwan

  • Luo, Kai-Hong;Chen, Ssu-Ching;Liao, Hung-Yu
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제21권5호
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    • pp.16-24
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    • 2016
  • A new strain of Pseudomonas sp. was isolated from mercury (Hg)-contaminated sites in Taiwan. This bacterium removed more than 80% of Hg present in the culture medium at 12 h incubation and was chosen for further analysis of the molecular mechanisms of Hg tolerance/removal abilities in this Pseudomonas sp. We used RNA-seq, one of the next-generation sequencing methods, to investigate the transcriptomic responses of the Pseudomonas sp. exposed to 60 mg/L of Hg2+. We de novo assembled 4,963 contigs, of which 10,533 up-regulated genes and 5,451 down-regulated genes were found to be regulated by Hg. The 40 genes most altered in expression levels were associated with tolerance to Hg stress and metabolism. Functional analysis showed that some Hg-tolerant genes were related to the mer operon, sulfate uptake and assimilation, the enzymatic antioxidant system, the HSP gene family, chaperones, and metal transporters. The transcriptome were analyzed further with Gene Ontology (GO) and Cluster of Orthologous Groups (COGs) of proteins and showed diverse biological functions and metabolic pathways under Hg stress.

Cloning of p-Hydroxybenzoate Degradation Genes and the Overexpression of Protocatechuate 4,5-Dioxygenase from Pseudomonas sp. K82

  • Yoon, Young-Ho;Park, Soon-Ho;Leem, Sun-Hee;Kim, Seung-Il
    • Journal of Microbiology and Biotechnology
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    • 제16권12호
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    • pp.1995-1999
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    • 2006
  • Pseudomonas sp. K82 cultured in p-hydroxybenzoate induces protocatechuate 4,5-dioxygenase (PCD 4,5) for p-hydroxybenzoate degradation. In this study, a 6.0-kbp EcoR1 fragment containing p-hydroxybenzoate degradation genes was cloned from the genome of Pseudomonas sp. K82. Sequence analysis identified four genes, namely, pcaD, pcaA, pcaB, and pcaC genes known to be involved in p-hydroxybenzoate degradation. Two putative 4-hydroxyphenylpyruvate dioxygenases and one putative oxidoreductase were closely located by the p-hydroxybenzoate degradation genes. The gene arrangement and sequences of these p-hydroxybenzoate degradation genes were similar to those of Comamonas testosteroni and Pseudomonas ochraceae. PcaAB (PCD4,5) was overexpressed in the expression vector pGEX-4T-3, purified using a GST column, and confirmed to have protocatechuate 4,5-dioxygenase activity. The N-terminal amino acid sequences of overexpressed PCD4,5 were identical with those of purified PCD4,5 from Pseudomonas sp. K82.

Pseudomonas sp. Aryl Acylamidase의 정제 및 성질 (Purification and Characterization of Aryl Acylamidase from Pseudomonas sp.)

  • 황인균;방원기
    • 한국미생물·생명공학회지
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    • 제26권5호
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    • pp.413-419
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    • 1998
  • Acetaminophen 자화성 Pseudomonas sp.에 존재하는 aryl acylamidase[EC 3.5.1.13]는 ammonium sulfate fractionation, DEAE-Sephacel anion exchange chromatography, Phenyl-Sepharose CL-4B hydrophobic interaction chroamtography 및 Sephadex G-100 gel-permeation chromatography를 통해 순수 정제되었다. 정제된 효소는 SDS-PAGE상에서 분자량을 측정한 결과 56 kDa, Sephadex G-100 gel-permeation chromatography로 측정한 결과 57 kDa이었으며, 따라서 단일한 subunit로 구성된 효소이었다. 정제된 효소의 최대 활성을 위한 pH와 온도는 각각 pH 10.5와 4$0^{\circ}C$이였다. 5$0^{\circ}C$에서 30분간 처리시 34%의 잔존활성을 나타내었다. Acetaminophen과 4'-nitroacetanilide쉐 대한 Km값은 각각 0.10 mM과 0.11 mM 이었다.

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Pseudomonas sp. Inulinase 유전자의 클로닝 및 Escherichia coli에서의 발현 (Molecular Cloning of Pseudomonas sp.Inulinase Gene and its Expresstion in E. coli)

  • 엄수정;권영만;최용진
    • 한국미생물·생명공학회지
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    • 제23권5호
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    • pp.550-555
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    • 1995
  • A strain of Pseudomonas sp. isolated from soil was shown to produce a high level of extracellular endo-inulinase. In this work, the endo-inulinase gene (inu1) of the bacterial strain was cloned into the plasmid pBR322 by using EcoRI restriction endonuclease and E. coli HB101 as a host strain. One out of 7, 000 transformants obtained from the above cloning experiment formed a clear zone around its colony on the selective medium supplemented with 2.0% inulin after a prolonged incubation at 37$\circ$C and subsequent cold shock treatment. The functional clone was found to carry a recombinant plasmid (pKMG50) with a 3.7 kb genomic insert containing the genetic information for the inulinase activity. The inulinase from E. coli HB101/pKMG50 was proved to be an endo-acting enzyme and produced constitutively in the recombinant E. coli cells. Zymogram of the enzyme from the recombinant cells with inulin substrate indicated that the molecular mass of the active protein was 190 Kd, while that of the endo-inulinase from the Pseudomonas strain was 170 Kd. This size discrepancy suggested that the inulinase from the recombinant E. coli HB101 cells might be the initial product of translation, not the mature form produced in the strain of Pseudomonas sp..

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저온.알칼리성 Protease를 생산하는 Pseudomonas sp. RP-222의 분리 및 조효소의 특성 (Isolation of Alkalopsychrotrophic Protease-Producing Pseudomonas sp. RP-222 and Properties of Its Crude Enzyme)

  • 노종수;정영철;성낙계;박석규
    • 한국미생물·생명공학회지
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    • 제19권4호
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    • pp.383-389
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    • 1991
  • 저온에서 높은 활성을 나타내는 알칼리성 protease를 생산하기 위하여 여러가지 시료로부터 집적배양에 의해 저온성 세균을 분리하였다. 분리된 세균은 저온.알칼리성 Pseudomonas sp.인 것으로 판명되었으며, 효소생산을 위한 균생육의 최적 pH는 10.0, 온도 $20^{\circ}C$에서 4일간 배양하였을 때였다. 이 효소 활성의 최적 pH 및 온도는 각각 pH 10.5 및 $40^{\circ}C$였으며, pH 및 열안정성은 각각 pH 7.0~13.0, 온도 $50^{\circ}C$이하의 범위에서 비교적 안정하였다.

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