2013
DOI: 10.1186/1754-1611-7-17
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Dual organism design cycle reveals small subunit substitutions that improve [NiFe] hydrogenase hydrogen evolution

Abstract: BackgroundPhotosynthetic microorganisms that directly channel solar energy to the production of molecular hydrogen are a potential future biofuel system. Building such a system requires installation of a hydrogenase in the photosynthetic organism that is both tolerant to oxygen and capable of hydrogen production. Toward this end, we have identified the [NiFe] hydrogenase from the marine bacterium Alteromonas macleodii “Deep ecotype” that is able to be heterologously expressed in cyanobacteria and has tolerance… Show more

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Cited by 13 publications
(32 citation statements)
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References 44 publications
(57 reference statements)
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“…In pIY107, the C-terminus of HynS is modified with a strep-tag [8,30,31] and the N-terminus of HynL is modified with a His 6 -tag. Details of the construction of pIY107 will be published elsewhere, but we provide its sequence in the Additional file 1.…”
Section: Methodsmentioning
confidence: 99%
See 2 more Smart Citations
“…In pIY107, the C-terminus of HynS is modified with a strep-tag [8,30,31] and the N-terminus of HynL is modified with a His 6 -tag. Details of the construction of pIY107 will be published elsewhere, but we provide its sequence in the Additional file 1.…”
Section: Methodsmentioning
confidence: 99%
“…The upstream amplicon was generated by primers BC000AvrF and BC001AvrR; the downstream amplicon was generated by primers BC002AvrF and IY171HynSR [8]; sequences and descriptions of primers are provided in Additional file 1: Table S4. A three-piece Gibson isothermal assembly was then performed using both amplicons and pIY107 doubly digested with AgeI and AvrII.…”
Section: Methodsmentioning
confidence: 99%
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“…The wild‐type enzyme favors hydrogen uptake activity over hydrogen evolution. However, a mutant with two substitutions in the Fe‐S cluster exhibited four times greater hydrogen evolution activity than the wild type .…”
Section: Optimization Of Hydrogen Production By the Genetic Modificatmentioning
confidence: 95%
“…Liang et al (2009) knocked out membrane-bound [NiFe] hydrogenase (MBH) in photosynthetic bacterium Allochromatium vinosum to improve the H 2 production rate. Similarly, H 2 evolution activities were increased by the mutation of hupW, which encodes putative hydrogenase-specific protease in Nostoc strain PCC 7120 of cyanobacterium (Lindberg, Devine, Stensj€ o, & Lindblad, 2011), inactivation of functional uptake hydrogenase (hupS) in nitrogen-fixing cyanobacterium Anabaena siamensis TISTR 8012 (Khetkorn et al, 2012), and substitution of amino acid, which alters the ligation of Fe-S clusters of [NiFe] uptake hydrogenase even in marine bacterium Alteromonas macleodii (Yonemoto, Matteri, Nguyen, Smith, & Weyman, 2013).…”
Section: Hydrogen Production By Photofermentationmentioning
confidence: 98%