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Relationship among growth parameters for Clostridium butyricum, hydA gene expression, and biohydrogen production in a sucrose-supplemented batch reactor

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Abstract

This study was undertaken to identify the relationship between the performance of dark H2 fermentation and expression of the key functional gene (i.e., hydrogenase gene) involved in the bioH2 production process. Clostridium butyricum CGS5 isolated from anaerobic sewage sludge was used as the model strain for this study. Copy number of the hydrogenase gene (hydA) and mRNA transcripts (cDNA hydA) (after amplification) and the total DNA and RNA (before amplification) were measured over the course of the growth of strain CGS5. Cell concentration was also determined by optical density and converted to dry weight. After amplification, the hydA gene increased 1,500-fold during late exponential growth phase after normalization to the copy number at time 0, and cDNA from mRNA transcripts of hydA also increased 500-fold after normalization. mRNA transcripts of hydA lagged behind the increase of total DNA and RNA, and increases in hydA more closely mimicked those of total DNA. Increases in both of these parameters corresponded with hydrogen production. Transcripts of 16s ribosomal RNA reached a maximum value earlier (38 h) than did those of hydA (47 h). All molecular characteristics matched those for sucrose utilization, growth, and hydrogen production. These experiments indicated that transcription as measured by cDNA can be related to hydrogen production and possesses the potential to be used as tool for process control.

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Acknowledgments

We thank the EISG grant 06-54324A/05-01 and the UC Pacific Rim Research Program grant 04-1299 for the funding to carry out this work. We also thank the financial support by Taiwan’s National Science Council grant 96-ET-7-006-004-ET and 094-2917-I-000-001.

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Correspondence to Betty H. Olson or Jo-Shu Chang.

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Wang, M., Olson, B.H. & Chang, J. Relationship among growth parameters for Clostridium butyricum, hydA gene expression, and biohydrogen production in a sucrose-supplemented batch reactor. Appl Microbiol Biotechnol 78, 525–532 (2008). https://doi.org/10.1007/s00253-007-1317-x

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Keywords

  • Biohydrogen
  • Clostridium butyricum
  • Expression of hydA gene
  • Hydrogenase
  • RT-PCR
  • qPCR