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| Construction of Rhodopseudomonas palustris hupL gene deficient mutant with higher efficient of H2 production. |
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Abstract Application of Rhodopseudomonas palustris CQU01 isolated from silt as initial strain to construct a hupL gene deficient mutant for promoting its H2-production. The amplified hupS and hupC in bilaterally of hupL of R. palustris were used as double-exchange arms in homologous recombination. They were inserted into pMD18-T for sequencing. The confirmed hupS, hupC and Kmr cassette were recombined into the pSUP202, constructing a suicide vector pBPZ for knocking-out the structure gene hupL of uptake hydrogenase. The pBPZ was identified based on enzyme digestion, amplification, sequencing and then transformed into R. palustris CQU01 via conjunction. A mutant strain was obtained by screened on Km culture plate and confirmed by PCR assay. The assay for total volume of H2 produced and growth patterns showed that the volume of H2 produced by the mutant strain was 50% higher than that of the wild strain of R. palustris and had similar growth patterns with the wild strain. It indicated that the hupL deficient mutant of R. palustris CQU012 may have a promising capability in the treatment of waste water.
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Received: 10 October 2008
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