Characterization and purification of a hydrogenase from the eukaryotic green alga <i>Scenedesmus obliquus</i>

Schnackenberg, Jörg; Schulz, Rüdiger; Senger, Horst

doi:10.1016/0014-5793(93)81030-4

Cited by 44 publications

(37 citation statements)

References 30 publications

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“…This geometry is consistent with efficient electron transfer among these prosthetic groups. As already shown in various studies, a correlation exists between the duration of time of the anaerobic adaptation and the increase of hydrogen production (30,32). Reverse transcription-PCR and Northern blot analyses with mRNA of aerobic and anaerobically adapted cells from S. obliquus showed an increased level of hydA transcript after 1 h of induction.…”

Section: Discussionsupporting

confidence: 66%

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A Novel Type of Iron Hydrogenase in the Green AlgaScenedesmus obliquus Is Linked to the Photosynthetic Electron Transport Chain

Florin

Tsokoglou

Happe

2001

Journal of Biological Chemistry

250

183

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Section: Discussionsupporting

confidence: 66%

“…In contrast, a nickel-iron hydrogenase was described for another well examined green alga, Scenedesmus obliquus (32,33). The protein consists of two subunits of about 36 and 55 kDa and might be located in the chloroplast.…”

mentioning

confidence: 98%

A Novel Type of Iron Hydrogenase in the Green AlgaScenedesmus obliquus Is Linked to the Photosynthetic Electron Transport Chain

Florin

Tsokoglou

Happe

2001

Journal of Biological Chemistry

250

183

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“…The activation energy value obtained for the acid phosphatase in this work is also closed to the values reported for other algae enzymes, such as hydrogenase (Schnackenberg et al, 1993) and urate oxidase (Alamillo et al, 1991) …”

Section: Kinetics Studiessupporting

confidence: 68%

Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata

Jonsson¹,

Aoyama

2009

Sci. agric. (Piracicaba, Braz.)

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Pseudokirchneriella subcapitata is a unicellular green algae widely distributed in freshwater and soils. Due to its cosmopolitan characteristic, its use is recommended by national and international protocols in ecotoxicity studies. The alteration of phosphatase activities by agriculture pollutants like heavy metals has been extensively used as a biomarker in risk assessment and biomonitoring. In this study, we compared the extraction of acid phosphatase from P. subcapitata by different procedures and we studied the stability, substrates specificity, kinetics and the effect of Hg 2+ in the crude extract. The freezing and thawing technique associated with probe sonication was the most suitable method of extraction. The enzyme was stable when frozen at -20ºC for at least six months, showed an optimum pH of 5 and a Km value of 0.27 mM for p-nitrophenylphosphate (pNPP) as substrate. Some natural organic substrates were cleaved by a similar extent as the synthetic substrate pNPP. RESUMO: Pseudokirchneriella subcapitata é uma alga verde unicelular amplamente distribuída em corpos d´agua e solos. Devido a sua natureza cosmopolita, seu uso é recomendado por protocolos nacionais e internacionais na realização de estudos de ecotoxicidade. A alteração da atividade de fosfatases por agentes poluentes de origem agrícola, como metais pesados, tem sido largamente usada como um biomarcador na avaliação de risco e biomonitoramento. No presente trabalho foi comparada a extração da fosfatase ácida de P. subcapitata por diferentes métodos e estudada a sua estabilidade, especificidade por substratos, cinética e efeito do Hg 2+ no extrato bruto. O congelamento e descongelamento, associado com ultrassom, foi o método que proporcionou maior rendimento de extração. A enzima, praticamente estável por armazenamento a -20ºC, durante aproximadamente seis meses, demonstrou uma atividade ótima em pH 5 e um valor de Km para o p-nitrofenilfostato (pNPP) de 0,27 mM. Alguns substratos naturais foram hidrolisados com uma intensidade semelhante à do substrato sintético pNPP. Diferentemente dos estudos de exposição a curto prazo (24 horas), observou-se inibição da atividade específica nas culturas expostas durante 7 dias, com um valor de CE 50 (concentração de Hg 2+ que promove 50% de decréscimo da atividade específica) equivalente a 12,63 μM Hg 2+ .

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“…Dihydrogen is produced by some green algae in the light under anaerobic conditions, e.g. in Chlamydomonas reinhardtii (Happe et al, 1994), and in Scenedesmus obliquus (Schnackenberg et al, 1993). Other lower eukaryotes that generate hydrogen gas include the microaerophilic protozoa (Yarlett et al, 1982) and chytrid fungi, e.g.…”

Section: Introductionmentioning

confidence: 99%

Giardia intestinalis, a eukaryote without hydrogenosomes, produces hydrogen

2002

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The microaerophilic flagellated protist Giardia intestinalis, the commonest protozoal agent of intestinal infections worldwide, is of uncertain phylogeny, but is usually regarded as the earliest branching of the eukaryotic clades. Under strictly anaerobic conditions, a mass spectrometric investigation of gas production indicated a low level of generation of dihydrogen (2 nmol min N1 per 10 7 organisms), about 10-fold lower than that in Trichomonas vaginalis under similar conditions. Hydrogen evolution was O 2 sensitive, and inhibited by 100 µM metronidazole. Fluorescent labelling of G. intestinalis cells using monoclonal antibodies to typical hydrogenosomal enzymes from T. vaginalis (malate enzyme, and succinyl-CoA synthetase α and β subunits), and to the large-granule fraction (hydrogenosome-enriched, also from T. vaginalis) gave no discrete localization of epitopes. Cell-free extracts prepared under anaerobic conditions showed the presence of a CO-sensitive hydrogenase activity. This first report of hydrogen production in a eukaryote with no recognizable hydrogenosomes raises further questions about the early branching status of G. intestinalis ; the physiological characterization of its hydrogenase, and its recently elucidated gene sequence, will aid further phylogenetic investigations.

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Characterization and purification of a hydrogenase from the eukaryotic green alga Scenedesmus obliquus

Cited by 44 publications

References 30 publications

A Novel Type of Iron Hydrogenase in the Green AlgaScenedesmus obliquus Is Linked to the Photosynthetic Electron Transport Chain

A Novel Type of Iron Hydrogenase in the Green AlgaScenedesmus obliquus Is Linked to the Photosynthetic Electron Transport Chain

Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata

Giardia intestinalis, a eukaryote without hydrogenosomes, produces hydrogen

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