The properties of hydrogenase from Thiocapsa roseopersicina

Гоготов, И. Н.; Зорин, Н. А.; Serebriakova, Larissa; Kondratieva, E. N.

doi:10.1016/0005-2744(78)90036-0

Cited by 83 publications

(25 citation statements)

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“…Thus, transfer of O 2 -tolerant [NiFe] hydrogenases into cyanobacteria might be one approach to overcome this O 2 sensitivity issue. A small number of O 2 -tolerant hydrogenases has been identified (9,21,47). However, they tend to favor H 2 uptake over evolution.…”

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confidence: 99%

“…In this study, we sought to prove the concept that metagenomically derived environmental DNA can give rise to a functional [NiFe] hydrogenase through expression in a foreign host and that novel [NiFe] hydrogenases from environmental microbes can be studied in a culture-independent manner. We cloned environmental DNA that harbors the genes of a putative novel hydrogenase that shows strong homology to a known O 2 -tolerant hydrogenase, HynSL, from the phototrophic purple sulfur bacterium Thiocapsa roseopersicina (21,28,41,59). We heterologously expressed the two structural genes (hyaA and hyaB) and two accessory genes (hupH and hyaD) of this novel environmental hydrogenase in T. roseopersicina, a foreign host that may already have the necessary machinery required to process the environmental hydrogenase since it carries the homologous hydrogenase HynSL.…”

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confidence: 99%

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Discovery of [NiFe] Hydrogenase Genes in Metagenomic DNA: Cloning and Heterologous Expression in Thiocapsa roseopersicina

Maróti

Ying-kai

Yooseph

et al. 2009

Appl Environ Microbiol

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Using a metagenomics approach, we have cloned a piece of environmental DNA from the Sargasso Sea that encodes an [NiFe] hydrogenase showing 60% identity to the large subunit and 64% to the small subunit of a Thiocapsa roseopersicina O 2 -tolerant [NiFe] hydrogenase. The DNA sequence of the hydrogenase identified by the metagenomic approach was subsequently found to be 99% identical to the hyaA and hyaB genes of an Alteromonas macleodii hydrogenase, indicating that it belongs to the Alteromonas clade. We were able to express our new Alteromonas hydrogenase in T. roseopersicina. Expression was accomplished by coexpressing only two accessory genes, hyaD and hupH, without the need to express any of the hyp accessory genes (hypABCDEF). These results suggest that the native accessory proteins in T. roseopersicina could substitute for the Alteromonas counterparts that are absent in the host to facilitate the assembly of a functional Alteromonas hydrogenase. To further compare the complex assembly machineries of these two [NiFe] hydrogenases, we performed complementation experiments by introducing the new Alteromonas hyaD gene into the T. roseopersicina hynD mutant. Interestingly, Alteromonas endopeptidase HyaD could complement T. roseopersicina HynD to cleave endoproteolytically the C-terminal end of the T. roseopersicina HynL hydrogenase large subunit and activate the enzyme. This study refines our knowledge on the selectivity and pleiotropy of the elements of the [NiFe] hydrogenase assembly machineries. It also provides a model for functionally analyzing novel enzymes from environmental microbes in a culture-independent manner.

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confidence: 99%

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confidence: 99%

Discovery of [NiFe] Hydrogenase Genes in Metagenomic DNA: Cloning and Heterologous Expression in Thiocapsa roseopersicina

Maróti

Ying-kai

Yooseph

et al. 2009

Appl Environ Microbiol

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“…The putative [NiFe] hydrogenase HynSL from AltDE displayed 60% identity to the large subunit and 64% to the small subunit of the stable [NiFe] hydrogenase (HynSL) from the purple sulfur bacterium Thiocapsa roseopersicina (12,20,30,54). In a previous study, we identified an Alteromonas [NiFe] hydrogenase from the Sargasso Sea, which is 99% identical to HynSL in AltDE (30).…”

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confidence: 99%

“…The T. roseopersicina [NiFe] hydrogenase HynSL has extraordinary stability at high temperatures (20). We determined whether A. macleodii hydrogenase HynSL is also thermostable.…”

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confidence: 99%

[NiFe] Hydrogenase from Alteromonas macleodii with Unusual Stability in the Presence of Oxygen and High Temperature

Vargas

Weyman

Ying-kai

et al. 2011

Appl Environ Microbiol

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Hydrogenases are enzymes involved in the bioproduction of hydrogen, a clean alternative energy source whose combustion generates water as the only end product. In this article we identified and characterized a [NiFe] hydrogenase from the marine bacterium Alteromonas macleodii "deep ecotype" with unusual stability toward oxygen and high temperature. The A. macleodii hydrogenase (HynSL) can catalyze both H 2 evolution and H 2 uptake reactions. HynSL was expressed in A. macleodii under aerobic conditions and reached the maximum activity when the cells entered the late exponential phase. The higher level of hydrogenase activity was accompanied by a greater abundance of the HynSL protein in the late-log or stationary phase. The addition of nickel to the growth medium significantly enhanced the hydrogenase activity. Ni treatment affected the level of the protein, but not the mRNA, indicating that the effect of Ni was exerted at the posttranscriptional level. Hydrogenase activity was distributed ϳ30% in the membrane fraction and ϳ70% in the cytoplasmic fraction. Thus, HynSL appears to be loosely membrane-bound. Partially purified A. macleodii hydrogenase demonstrated extraordinary stability. It retained 84% of its activity after exposure to 80°C for 2 h. After exposure to air for 45 days at 4°C, it retained nearly 100% of its activity when assayed under anaerobic conditions. Its catalytic activity in the presence of O 2 was evaluated by the hydrogen-deuterium (H-D) exchange assay. In 1% O 2 , 20.4% of its H-D exchange activity was retained. The great stability of HynSL makes it a potential candidate for biotechnological applications.

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“…A T. roseopersicina BBS egy, a Chromatiaceae családba tartozó anaerob fototróf, bíbor kénbaktérium (Bogorov 1974, Gogotov 1976, Pfennig 1983. Az 1-3 μm átmérőjű, nem mozgékony sejtek számára optimális növekedési hőmérséklet 24-28°C között van (Bogorov 1974).…”

Section: Thiocapsa Roseopersicinaunclassified

Fotobiohidrogén termelésében szerepet játszó enzimek bioszintézise és metabolikus kontextusa

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The properties of hydrogenase from Thiocapsa roseopersicina

Cited by 83 publications

References 10 publications

Discovery of [NiFe] Hydrogenase Genes in Metagenomic DNA: Cloning and Heterologous Expression in Thiocapsa roseopersicina

Discovery of [NiFe] Hydrogenase Genes in Metagenomic DNA: Cloning and Heterologous Expression in Thiocapsa roseopersicina

[NiFe] Hydrogenase from Alteromonas macleodii with Unusual Stability in the Presence of Oxygen and High Temperature

Fotobiohidrogén termelésében szerepet játszó enzimek bioszintézise és metabolikus kontextusa

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