Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

M. C. Posewitz; P. W. King; S. L. Smolinski; R. Davis Smith; A. R. Ginley; M. L. Ghirardi; M. Seibert

doi:10.1042/BST0330102

Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

M. C. Posewitz
, P. W. King
, S. L. Smolinski
, R. Davis Smith
, A. R. Ginley
, M. L. Ghirardi
, M. Seibert

National Renewable Energy Laboratory
Colorado School of Mines

Research output: Contribution to journal › Article › peer-review

83 Scopus Citations

Abstract

The eukaryotic green alga, Chlamydamonas reinhardtii, produces H ₂ under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H₂ production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H₂-sensor films for clones defective in H ₂ production. Two mutants of particular interest were fully characterized. One mutant, hydEF-1, is unable to assemble an active [FeFe]-hydrogenase. This is the first reported C. reinhardtii mutant that is not capable of producing any H₂. The second mutant, sta7-10, is not able to accumulate insoluble starch and has significantly lowered H ₂-photoproduction rates in comparison with the wild-type. In hydEF-1, anaerobiosis induces transcription of the two reported C. reinhardtii hydrogenase genes, HydA1 and HydA2, indicating a normal transcriptional response to anaerobiosis. In contrast, the transcription of both hydrogenase genes in sta7-10 is significantly attenuated.

Original language	American English
Pages (from-to)	102-104
Number of pages	3
Journal	Biochemical Society Transactions
Volume	33
Issue number	1
DOIs	https://doi.org/10.1042/BST0330102 https://doi.org/10.1042/BST0330102
State	Published - 2005

NLR Publication Number

NREL/JA-590-38657

Keywords

Chlamydomonas reinhardtii
H photoproduction
Hydrogenase
Insertional mutagenesis
Metalloprotein assembly
Starch

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title = "Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii",

abstract = "The eukaryotic green alga, Chlamydamonas reinhardtii, produces H 2 under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H2 production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H2-sensor films for clones defective in H 2 production. Two mutants of particular interest were fully characterized. One mutant, hydEF-1, is unable to assemble an active [FeFe]-hydrogenase. This is the first reported C. reinhardtii mutant that is not capable of producing any H2. The second mutant, sta7-10, is not able to accumulate insoluble starch and has significantly lowered H 2-photoproduction rates in comparison with the wild-type. In hydEF-1, anaerobiosis induces transcription of the two reported C. reinhardtii hydrogenase genes, HydA1 and HydA2, indicating a normal transcriptional response to anaerobiosis. In contrast, the transcription of both hydrogenase genes in sta7-10 is significantly attenuated.",

keywords = "Chlamydomonas reinhardtii, H photoproduction, Hydrogenase, Insertional mutagenesis, Metalloprotein assembly, Starch",

author = "Posewitz, \{M. C.\} and King, \{P. W.\} and Smolinski, \{S. L.\} and Smith, \{R. Davis\} and Ginley, \{A. R.\} and Ghirardi, \{M. L.\} and M. Seibert",

year = "2005",

doi = "10.1042/BST0330102",

language = "American English",

volume = "33",

pages = "102--104",

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T1 - Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

AU - Posewitz, M. C.

AU - King, P. W.

AU - Smolinski, S. L.

AU - Smith, R. Davis

AU - Ginley, A. R.

AU - Ghirardi, M. L.

AU - Seibert, M.

PY - 2005

Y1 - 2005

N2 - The eukaryotic green alga, Chlamydamonas reinhardtii, produces H 2 under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H2 production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H2-sensor films for clones defective in H 2 production. Two mutants of particular interest were fully characterized. One mutant, hydEF-1, is unable to assemble an active [FeFe]-hydrogenase. This is the first reported C. reinhardtii mutant that is not capable of producing any H2. The second mutant, sta7-10, is not able to accumulate insoluble starch and has significantly lowered H 2-photoproduction rates in comparison with the wild-type. In hydEF-1, anaerobiosis induces transcription of the two reported C. reinhardtii hydrogenase genes, HydA1 and HydA2, indicating a normal transcriptional response to anaerobiosis. In contrast, the transcription of both hydrogenase genes in sta7-10 is significantly attenuated.

AB - The eukaryotic green alga, Chlamydamonas reinhardtii, produces H 2 under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H2 production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H2-sensor films for clones defective in H 2 production. Two mutants of particular interest were fully characterized. One mutant, hydEF-1, is unable to assemble an active [FeFe]-hydrogenase. This is the first reported C. reinhardtii mutant that is not capable of producing any H2. The second mutant, sta7-10, is not able to accumulate insoluble starch and has significantly lowered H 2-photoproduction rates in comparison with the wild-type. In hydEF-1, anaerobiosis induces transcription of the two reported C. reinhardtii hydrogenase genes, HydA1 and HydA2, indicating a normal transcriptional response to anaerobiosis. In contrast, the transcription of both hydrogenase genes in sta7-10 is significantly attenuated.

KW - Chlamydomonas reinhardtii

KW - H photoproduction

KW - Hydrogenase

KW - Insertional mutagenesis

KW - Metalloprotein assembly

KW - Starch

UR - https://www.scopus.com/pages/publications/14644393653

U2 - 10.1042/BST0330102

DO - 10.1042/BST0330102

M3 - Article

C2 - 15667277

AN - SCOPUS:14644393653

SN - 0300-5127

VL - 33

SP - 102

EP - 104

JO - Biochemical Society Transactions

JF - Biochemical Society Transactions

IS - 1

ER -

Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

Abstract

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Keywords

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