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Published online ahead of print on 16 April 2009 as doi:10.1099/mic.0.025536-0
Microbiology 2009;155:1717.

Microbiology (2009), DOI 10.1099/mic.0.025536-0
© 2009 Society for General Microbiology
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Microbiology 0 (2009), mic.0.025536; DOI  10.1099/mic.0.025536-0
© 2009 Society for General Microbiology

Systematic characterization of a novel gal operon in Thermoanaerobacter tengcongensis

Z. Qian1, B. Meng1, Q. Wang2, Z. Wang1, C. Zhou1, Q. Wang1, S. Tu1, L. Lin1, Y. Ma2 and S. Liu3,4

1 Beijing Institute of Genomics;
2 Institute of Microbiology;
3 Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China

ABSTRACT

On the basis of Thermoanaerobacter tengcongensis genome, a novel type of gal operon was deduced. The gene expression and biochemical properties of this operon were further characterized. RT-PCR analysis of the intergenic regions suggested that the transcription of gal operon was continuous. With gene cloning and enzyme activity assays, TTE1929, TTE1928 and TTE1927 were identified to be GalT, GalK and GalE, respectively. Results elicited from polarimeter assay revealed that TTE1925, a hypothetical protein, was a novel mutarotase, termed as MR-Tt. TTE1926 was identified as the regulator which could bind to two operators in the operon promoter. The transcriptional start sites were mapped, which suggested that two promoters existed in this operon. Expression of the gal genes was significantly induced by galactose, whereas only MR-Tt expression was detected in the glucose cultured T. tengcongensis at both mRNA and protein levels. Moreover, the abundance of gal proteins was examined at different temperatures. At temperatures ranging from 60 to 80 °C, the protein content of MR-Tt was relatively stable, but that of the other gal proteins was dramatically decreased. The operator-binding complexes were isolated and identified by EMSA-LC MS/MS, implying that several regulation proteins, such as GalR and sensory histidine kinase, might participate in the regulation of gal operon.

4 E-mail: siqiliu{at}genomics.org.cn






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