• 제목/요약/키워드: pyrH

검색결과 24건 처리시간 0.021초

젖산탈수소효소 eye-specific C4 동위효소의 생화학적 특성: 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides) (Biochemical Properties of Lactate Dehydrogenase Eye-Specific C4 Isozyme: Lepomis macrochirus and Micropterus salmoides)

  • 염정주;구보라
    • 생명과학회지
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    • 제22권2호
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    • pp.209-219
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    • 2012
  • 눈조직의 젖산탈수소효소(LDH, EC 1.1.1.27) eye-specific $C_4$ 동위효소의 특성을 native-PAGE, Western blotting, 면역침강반응 및 효소 역학을 이용하여 연구하였고, LDH eye-specific $C_4$ 동위효소의 활성을 측정하는 조건을 제시하였다. 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides) 눈조직의 세포기질에서 LDH eye-specific $C_4$ 동위효소가 확인되었으며 $B_4$ 동위효소보다 $A_4$ 동위효소에 유사하였다. 파랑볼우럭 눈조직의 LDH/CS는 9월에 증가하여 혐기적 대사가 높게 이루어졌다. 파랑볼우럭과 큰입우럭 눈조직 미토콘드리아 LDH의 전기영동상은 세포기질 LDH와 유사하였다. 눈조직의 LDH eye-specific $C_4$ 동위효소는 Preparative native-PAGE에 의해 정제되었다. 파랑볼우럭과 큰입우럭의 LDH eye-specific $C_4$ 동위효소의 활성은 피루브산 0.2 mM과 0.1 mM 이상의 농도에서 각각 감소되었고, 피루브산 10 mM에서 5.2%, 15.8% 활성이 남아 저해 정도가 크게 나타났다. 그리고 눈조직의 LDH 활성도 젖산 22 mM과 24 mM 이상의 농도에서 각각 감소되어졌다. 파랑 볼우럭 eye-specific $C_4$ 동위효소의 $Km^{PYR}$는 0.088 mM, 큰입우럭 eye-specific $C_4$ $Km^{PYR}$는 0.033 mM였으며, 세포기질과 미토콘드리아 eye-specific $C_4$ 동위효소는 ${\alpha}$-ketobutyric acid에서 활성이 높게 나타났다. 눈조직과 eye-specific $C_4$ 동위효소의 활성은 피루브산 0.5 mM과 Tris-HCl 완충액, pH 7.5에서 측정하는 것이 적합한 것으로 확인되었다. 실험 결과, 큰입우럭 LDH eye-specific $C_4$ 동위효소는 파랑볼우럭 LDH eye-specific $C_4$보다 피루브산에 대한 친화력이 큰 것으로 확인되었고, 더 낮은 피루브산의 농도에서 최대 활성에 이르고, 더 높은 젖산의 농도에서 최대 활성을 나타냈다. 따라서 LDH eye-specific $C_4$ 동위효소에 의해 생성된 에너지가 포식 행동의 초기 반응에 사용되는 것으로 사료된다.

Assessment of Reliability when Using Diagnostic Binary Ratios of Polycyclic Aromatic Hydrocarbons in Ambient Air PM10

  • Pongpiachan, Siwatt
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권18호
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    • pp.8605-8611
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    • 2016
  • The reliability of using diagnostic binary ratios of particulate carcinogenic polycyclic aromatic hydrocarbons (PAHs) as chemical tracers for source characterisation was assessed by collecting PM10 samples from various air quality observatory sites in Thailand. The major objectives of this research were to evaluate the effects of day and night on the alterations of six different PAH diagnostic binary ratios: An/(An + Phe), Fluo/(Fluo + Pyr), B[a]A/(B[a]A + Chry), B[a]P/(B[a]P + B[e]P), Ind/(Ind + B[g,h,i]P), and B[k]F/Ind, and to investigate the impacts of site-specific conditions on the alterations of PAH diagnostic binary ratios by applying the concept of the coefficient of divergence (COD). No significant differences between day and night were found for any of the diagnostic binary ratios of PAHs, which indicates that the photodecomposition process is of minor importance in terms of PAH reduction. Interestingly, comparatively high values of COD for An/(An + Phe) in PM10 collected from sites with heavy traffic and in residential zones underline the influence of heterogeneous reactions triggered by oxidising gaseous species from vehicular exhausts. Therefore, special attention must be paid when interpreting the data of these diagnostic binary ratios, particularly for cases of low-molecular-weight PAHs.

명태(Theragra chalcogramma) 종묘에서 분리된 Vibrio anguillarum의 생화학적, 분자생물학적 특성 (Biochemical and Molecular Characteristics of Vibrio anguillarum Isolated from Alaska Pollock Theragra chalcogramma Seedlings)

  • 전찬혁;남우화;김정호
    • 한국수산과학회지
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    • 제49권3호
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    • pp.359-366
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    • 2016
  • The health of Alaska pollock Theragra chalcogramma seedlings was monitored during February and April 2015. After microscopic examination for parasites, 20 samples sets were made by pooling 50 individuals for each sample set. Then, they were homogenized and examined for viral and bacterial pathogens. No parasites or viruses were detected using either microscopy or PCR. Colonies suspected of belonging to the genus Vibrio were isolated from Tryptic Soya Agar and Thiosulfate Citrate Bile Salts Sucrose Agar plate incubations, and identified as Vibrio anguillarum based on biochemical and physiological examinations and PCR amplification of the 16S rDNA, recA, and pyrH genes. Although there was no mortality during the sampling period, 65.0% (13/20) of the pooled samples were PCR-positive for V. anguillarum. To prevent possible outbreaks, the pathogenic potential of V. anguillarum should be investigated in the future.

Phytotoxin Production of Nigrospora sphaerica Pathogenic on Turfgrasses

  • Park, Gyung-Ja;Kim, Jin-Cheol;Shon, Mi-Jeong;Kim, Heung-Tae;Cho, Kwang-Yun
    • The Plant Pathology Journal
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    • 제16권3호
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    • pp.137-141
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    • 2000
  • A causal fungus of turfgrass blight was isolated from the infected leaves of zoysiagrass (Zoysia japonica Steud.) and identified as Nigrospora sphaerica (Sacc.) Mason by using a light misroscope. Its conidia are large (14-20 ${\mu}{\textrm}{m}$ diameter), shiny, black, aseptate, and smooth-walled spheres. The fungus caused typical blighting symptoms on the two turfgrass plants of bermudagrass (Cynodon dactylon (L.) Pers.) and bentgrass (Agrostis palustris Huds.). The fungus was found to produce a phytotoxic subtance to be associated with the pathogenic mechanism. A phytotoxin was isolated from the liquid cultures of N. sphaerica by repeated silica gel column chromatography and its structure was determined to be 5, 6-dihydro-5-hydroxy-6-propenyl-2H-pyr-2-one (T-3 compound). It was not a host-specific toxin showing phytotoxic effects to various plants inclusing turfgrasses in the leaf-wounding assay, the whole plant test, and the cellular leakage test. The compound caused leaf tip dieback symptoms in turfgrass plants similar to those caused by the pathogen. Thus, T-3 compound is thought to be involved in the development of Nigrospora blight.

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가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성 (Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus)

  • 박은미;염정주
    • 생명과학회지
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    • 제20권2호
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    • pp.260-268
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    • 2010
  • 가물치(Channa argus) 조직의 젖산탈수소효소 동위효소(EC 1.1.1.27, lactate dehydrogenase, LDH)를 정제하고 생화학적, 면역학적 및 역학적 방법으로 특성을 연구하였다. LDH 활성은 골격근이 380.4 units로 가장 높고 심장 13.4, 눈 3.5, 뇌 조직 5.4 units이었으며, 심장의 CS 활성은 20.7 unit로 가장 높고, LDH/CS는 골격근 172.9, 심장 0.6, 눈 0.32, 뇌 0.47이고, 단백질 양은 골격근 14.7 mg/g이며, 특이활성(units/mg)은 골격근 25.88, 심장 0.79, 눈 0.31, 뇌 1.38 units/mg이었으므로 골격근은 혐기적이고, 심장은 호기적이었다. LDH $A_4$, $B_4$, eye-specific $C_4$에 대한 항혈청을 사용한 Western blot, 면역침강반응 및 native-polyacrylamide gel electrophoresis에 의해 $A_4$, $A_3B$, $A_2B_2$, $AB_3$$B_4$가 모든 조직에서 확인되었고, 눈 조직에서 $C_4$$AC_3$, $A_2C_2$, $AC_3$, 뇌 조직에서 $A_3C$도 확인되었다. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ 동위효소는 affinity chromatography와 Preparative PAGE Cell에 의해 정제되었다. LDH $A_4$ 동위효소는 $NAD^+$ 유입 후 정제되었고, eye-specific $C_4$$A_4$에 이어 용출되기 시작하였으며 $B_4$는 buffer 유입 후 용출되었다. 정제한 결과 $A_4$$B_4$ 및 eye-specific $C_4$와 분자구조의 일부가 유사하였지만 $B_4$$C_4$는 서로 다른 것으로 나타났으므로, 하부단위체 A는 보존적이고, 하부단위체 B는 A보다 더 빠르게 진화된 것으로 보인다. 피루브산 10 mM에서 $A_4$ 동위효소 39.98%, $A_2B_2$ 21.28%, $B_4$ 19.67% 및 eye-specific $C_4$ 16.87%의 활성이 남아있었고, 피루브산에 대한 $Km^{PYR}$$A_4$ 0.17 mM, $B_4$ 0.27 mM, eye-specific $C_4$ 0.133 mM였다. $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$$AB_3$의 최적 pH는 각각 pH 6.50, pH 8.5, pH 5.5, pH 6.0-6.5, 5.0 및 pH 7.5였고, 동질사량체 $A_4$와 이질사량체 동위효소들은 넓은 pH 영역에서 안정하였다. 특히 골격근은 LDH 활성이 크므로 활동성이 크며, 눈조직에서 피루브산 친화력이 강한 eye-specific $C_4$에 의해 피루브산 대사가 빠르게 일어나고, 이어서 $A_4$에 의해 젖산이 산화되어지는 것으로 사료되므로, 종의 생태환경 및 먹이 획득 양식에 따라 LDH-C 발현, 기질에 대한 친화도 및 대사 시간이 다른 것으로 사료된다.

The High Performance Liquid Chromatography (HPLC) Analysis of Polycyclic Aromatic Hydrocarbons (PAHs) in Oysters from the Intertidal and Subtidal Zones of Chinhae Bay, Korea

  • Lee Ki Seok;Noh Il;Lim Cheol Soo;Chu Su Dong
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • 제2권1호
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    • pp.57-68
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    • 1998
  • Polycyclic aromatic hydrocarbons (PAMs) are ubiquitous contaminants in marine environments. PAMs enter estuarine and nearshore marine environment via several routes such as combustion of fossil fuels, domestic and industrial effluents and oil spills. PAHs have been the focus of numerous studies in the world because they are potentially carcinogenic, mutagenic, and teratogenic to aquatic organisms and humans from consuming contaminated food. However, one can hardly find any available data on PAM content in marine organisms in Korea. The present study was carried out in order to determine PAM content in oysters from the intertidal and subtidal zones of Chinhae Bay, which is located in near urban communities and an industrial complex, and the bay is considered to be a major repositories of PAHs. 16 PAHs were analyzed by High Performance Liquid Chromatography (HPLC) with uv/vis and fluorescence detectors in oysters: they are naphthalene (NPTHL), acenaphthylene (ANCPL), acenaphthylene (ACNPN), fluorene (FLURN), phenanthrene (PHEN), anthracene (ANTHR), fluoranthene (FLRTH), pyrene (PYR), benzo(a)anthracene (BaA), chrysene (CHRY), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP), dibenz(a,h)anthracene(DahA), benzo(g,h,i)peryne (BghiP) and indeno(1,2,3,-cd)pyrene (I123cdP). The PAM contents in oysters from the intertidal and subtidal zones of Chinhae Bay ranged from < 0.1 to 992.0 ug/kg (mean $69.8\pm9.8$ ug/kg).

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큰입우럭(Micropterus salmoides) 조직의 젖산탈수소효소 및 Monocarboxylate 수송체(MCT) 1, 2, 4 (Lactate Dehydrogenase and Monocarboxylate Transporters 1, 2, and 4 in Tissues of Micropterus salmoides)

  • 염정주;연준희
    • 생명과학회지
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    • 제22권1호
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    • pp.98-109
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    • 2012
  • 큰입우럭(Micropterus salmoides) 조직의 젖산탈수소효소(EC 1.1.1.27, LDH)의 특성 및 골격근과 심장조직의 monocarboxylate 수송체 1, 2, 4의 발현을 연구하였다. Native-PAGE 결과골격근에서 LDH $A_4$, 심장, 간, 눈 및 뇌조직에서 $A_4$, $A_2B_2$, $B_4$, 눈조직에서 eye-specific $C_4$ 동위효소가 발현되었다. 9월에 심장조직에서 LDH $B_4$ 동위효소의 활성이 강하고 다른 조직에서는 $A_4$의 활성이 강했으나, 11월에 심장조직에서 $A_4$ 동위효소의 활성이 강하게 확인되었다. 골격근과 심장조직에서 LDH/CS로 조직의 혐기적 대사 비율이 높게 확인되었으며, 면역 침강 후 native-PAGE에 의해 LDH eye-specific $C_4$ 동위효소가 $B_4$보다 $A_4$ 동위효소에 더 유사한 것으로 확인되었다. LDH $A_4$ 동위효소가 affinity chromatography에 의해 정제되었고, 하부단위체 A의 분자량은 37.200이었다. 피루브산 10 mM에서 조직 LDH의 활성이 11.05-28.32% 남아 저해 정도가 컸고, 눈조직 LDH의 $K_m^{PYR}$이 낮았으며, 조직의 최적 pH는 7.5~8.0이였다. 골격근 미토콘드리아에서 LDH $A_4$ 동위효소, 심장조직의 미토콘드리아에서 $B_4$$A_2B_2$ 동위효소가 확인되었고, 골격근과 심장조직의 원형질막과 미토콘드리아에서 MCT 1, 2, 4가 확인되었다. 골격근 MCT 1, 2, 4 골격근 MCT 1 60 kDa, MCT 2 54~38 kDa, MCT 4 63 kDa, 심장조직 MCT 1 57 kDa, MCT 2 54~38 kDa 및 MCT 4 55.5 kDa이었다. 실험 결과, 큰입우럭이 저산소 조건에 적응되어져 혐기적 대사가 우세하고, 활성이 큰 골격근과 심장조직에서 원형질막과 미토콘드리아 MCT 1, 2, 4를 통해 젖산과 피루브산이 유입되고 유출되며 LDH에 의해 에너지 생성을 효율적으로 조정하는 것으로 사료된다.

마늘의 alcohol 침지 중 휘발성 향기성분과 침출유리아미노산 함량 (Elution Profiles of Volatile Compounds and Free Amino Acids during Alcohol Soaking of Garlic(Allum sativum L.))

  • 이영근
    • 생명과학회지
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    • 제17권2호통권82호
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    • pp.286-292
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    • 2007
  • 마늘주의 산업화에 앞서 기초 자료로써 활용하고자, 마늘을 소주 등의 술에 침지하였을 때 마늘의 휘발성 기능성분과 유리아미노산이 술로 침출되는 정도 및 형태에 관하여 조사하고자, 생마늘을 20% 주정에 5주간 침지하고 매주 휘발성 향기성분과 유리아미노산들이 침출되는 양적 및 형태적 변화를 조사하였다. 마늘의 주요 유리 아미노산은 L-aspartic acid, L-glutamic acid, L-arginine, L-alanine, L- proline, L-asparagine 및 L-serine이었으며, 유리 아미노산들 모두 침지 기간의경과에 따라 침출량이 증가하였다. 또한, L-threonine, L-proline, L-valine 및 L-leucine 등의 중성아미노산들과 방향족아미노산인 tyrosine과 phenylalanine은 3주 후부터 80% 이상 침출된 반면에, 산성, 염기성 및 함황아미노산들은 5주 후에도 80% 미만이었다. 생마늘의 휘발성 향기성분은 diallyl trisulfide, diallyl disulfide, methyl allyl disulfide, 2-vinyl-4H-1,3-dithiin, 3-vinyl-3,4-dihydro-1,2-dithiin, 3,5-diethyl-1,24-trithiolane, isobutyl isothiocyanate 및 diallyl sulfide 등의 황화합물들이 주요 성분들이었다. 생마늘로부터 주정으로 침출된 휘발성 향기성분은 allyl alcohol, diallyl disulfide, 3,5-diethyl-1,2,4-trithiolane, diallyl trisulfide, 3,4-dimethoxyfuran의 5종이었으며, 침지기간의 경과에 따라 3,5-diethyl-1,2,4-trithiolane는 계속 침출량이 증가하였지만 그 외 4종은 감소하였다. 침 지기간 중 furfural, 5-methylfurfural, 5-hydroxymethylfurfural, dimethyl pyrazine, furfuryl alcohol, 3-hydroxy-2-bytanone과 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one이 새로이 생성되었으며, 침지기간의 경과에 따라 침출주에 대부분 축적되는 양상을 보였다.

Multilocus sequence analysis of the genus Aliivibrio: Identification and phylogeny of Aliivibrio species isolated from cultured walleye pollock (Gadus chalcogrammus) in Korea

  • Nam, U-Hwa;Seo, Hyun-Joon;Jang, Su-Rim;Kim, Mi-Ri;Kim, Jeong-Ho
    • 한국어병학회지
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    • 제32권2호
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    • pp.69-80
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    • 2019
  • We performed MLSA (multilocus sequence analysis) and phenotypic characterization of Aliivibrio species isolated from walleye pollock (Gadus chalcogrammus) maintained in 3 different facilities of Gangwon Province, the east coast of Korea. Of 38 Aliivibrio species identified by 16S rDNA sequences, 12 strains were randomly selected and MLSA was conducted with 5 house-keeping genes (gapA, gyrB, pyrH, recA and rpoA) and 16S rDNA gene. Phylogenetic analysis and homology of the concatenated sequences (4,580 bp) with other Vibrionaceae genera revealed that 4 strains (GNGc16.1, YYGc16.1, YYGc16.2, GSGc18.1) were identified as Aliivibrio logei and one strain (GSGc16.1) as A. wodanis. One strain (GSGc17.1) was tentatively identified as A. logei, but needs further analysis because it did not belong to the same clade with A. logei type strain. 6 strains (GSGc17.2, GNGc16.2, GSGc16.2, GSGc17.3, GSGc18.2, GSGc17.4) need further investigation as potential novel species. Either phenotypic characterization or 16S rDNA sequence alone did not provide enough information for identification of Aliivibrio strains at the species level. A. logei and A. wodanis are generally known as non-pathogenic bacteria, but also known as opportunistic or secondary pathogens of cold water fishes. Cares should be taken to prevent potential outbreaks due to these bacteria, although there was no outbreaks during the sampling period.

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal α-Amylase Gene in Aspergillus oryzae

  • Yin, Yanchen;Mao, Youzhi;Yin, Xiaolie;Gao, Bei;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • 제25권7호
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    • pp.988-998
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    • 2015
  • The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.