Translational and transcriptional analysis of <b><i>Sulfolobus solfataricus</i></b> P2 to provide insights into alcohol and ketone utilisation

Chong, Poh Kuan; Burja, Adam M.; Radianingtyas, Helia; Fazeli, Alireza; Wright, Phillip C.

doi:10.1002/pmic.200600746

Cited by 20 publications

(45 citation statements)

References 53 publications

(39 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In S. solfataricus, besides Sso2536, 12 additional open reading frames (ORFs) encoding putative ADHs have been identified and annotated (adh-1 to -13). All but adh-9 are actively expressed, as demonstrated by proteome analyses (17,18). The presence of multiple ADH ORFs indicates a metabolic relevance of alcohol metabolism by this organism (16).…”

mentioning

confidence: 99%

MarR-Like Transcriptional Regulator Involved in Detoxification of Aromatic Compounds in Sulfolobus solfataricus

et al. 2007

View full text Add to dashboard Cite

A DNA binding protein, BldR, was identified in the crenarchaeon Sulfolobus solfataricus as a protein 5-to 10-fold more abundant in cells grown in the presence of toxic aldehydes; it binds to regulatory sequences located upstream of an alcohol dehydrogenase gene (Sso2536). BldR is homologous to bacterial representatives of the MarR (multiple antibiotic resistance) family of transcriptional regulators that mediate response to multiple environmental stresses. Transcriptional analysis revealed that the bldR gene was transcribed in a bicistronic unit composed of the genes encoding the transcriptional regulator (Sso1352) and a putative multidrug transporter (Sso1351) upstream. By homology to bacterial counterparts, the bicistron was named the mar-like operon. The level of mar-like operon expression was found to be increased at least 10-fold in response to chemical stress by aromatic aldehydes. Under the same growth conditions, similar enhanced in vivo levels of Sso2536 gene transcript were also measured. The gene encoding BldR was expressed in E. coli, and the recombinant protein was purified to homogeneity. DNA binding assays demonstrated that the protein is indeed a transcription factor able to recognize site specifically both the Sso2536 and mar-like promoters at sites containing palindromic consensus sequences. Benzaldehyde, the substrate of ADH Ss , stimulates DNA binding of BldR at both promoters. The role of BldR in the auto-activation as well as in the regulation of the Sso2536 gene, together with results of increased operon and gene expression under conditions of exposure to aromatic aldehydes, indicates a novel coordinate regulatory mechanism in cell defense against stress by aromatic compounds.A chimeric nature of the transcription machinery with eukaryote-like basal factors and bacterium-like regulative components has been found in all members of the domain Archaea (50, 60). In fact, in most cases, homologs of bacterial regulators function in the context of the archaeal basal transcriptional apparatus, which resembles that of Eukarya (8, 26). Transcription initiation is mediated by a single RNA polymerase (RNAP) and two general transcription factors, TATA element binding protein (TBP) and transcription factor B (TFB), a homologue of the transcription factor TFIIB, which binds to the B recognition element (BRE) and determines the directionality of the transcription (7). The complex containing RNAP, TBP, and TFB is sufficient to initiate transcription in cell-free systems (31, 49), although an additional factor, transcription factor E, is required to increase transcription from some promoters (6, 29).A few homologs of eukaryal transcriptional regulators (33) and unique archaeal regulators (57) as well as non-sequencespecific DNA binding proteins (5, 30) have been investigated for their contribution to the regulation of archaeal genes.Homologs of bacterial transcriptional regulators are more common in Archaea, and representatives that have been studied experimentally include the Lrp homologs (13) that can fun...

show abstract

mentioning

confidence: 99%

MarR-Like Transcriptional Regulator Involved in Detoxification of Aromatic Compounds in Sulfolobus solfataricus

et al. 2007

View full text Add to dashboard Cite

show abstract

“…Comparative lipidomics analysis has been applied to study the ethanologenic yeast response to different inhibitors, such as furfural, acetic acid, and phenol (69). The results of comparative proteome analysis (8,23) and small-molecule metabolite profiling of ethanologenic yeast during industrial fermentation (13) have been previously reported, enhancing the molecular understanding of physiological adaptation of industrial strains for optimizing the performance of industrial bioethanol fermentation. However, the specifics of global protein expression in response to the presence of biomass conversion inhibitors have not yet been quantitatively measured for ethanologenic yeast.…”

mentioning

confidence: 99%

Comparative Proteomic Analysis of Tolerance and Adaptation of EthanologenicSaccharomyces cerevisiaeto Furfural, a Lignocellulosic Inhibitory Compound

Lin

Qiao

Yang

2009

Appl Environ Microbiol

100

View full text Add to dashboard Cite

The molecular mechanism involved in tolerance and adaptation of ethanologenic Saccharomyces cerevisiae to inhibitors (such as furfural, acetic acid, and phenol) represented in lignocellulosic hydrolysate is still unclear. Here, 18 O-labeling-aided shotgun comparative proteome analysis was applied to study the global protein expression profiles of S. cerevisiae under conditions of treatment of furfural compared with furfural-free fermentation profiles. Proteins involved in glucose fermentation and/or the tricarboxylic acid cycle were upregulated in cells treated with furfural compared with the control cells, while proteins involved in glycerol biosynthesis were downregulated. Differential levels of expression of alcohol dehydrogenases were observed. On the other hand, the levels of NADH, NAD ؉ , and NADH/NAD ؉ were reduced whereas the levels of ATP and ADP were increased. These observations indicate that central carbon metabolism, levels of alcohol dehydrogenases, and the redox balance may be related to tolerance of ethanologenic yeast for and adaptation to furfural. Furthermore, proteins involved in stress response, including the unfolded protein response, oxidative stress, osmotic and salt stress, DNA damage and nutrient starvation, were differentially expressed, a finding that was validated by quantitative real-time reverse transcription-PCR to further confirm that the general stress responses are essential for cellular defense against furfural. These insights into the response of yeast to the presence of furfural will benefit the design and development of inhibitor-tolerant ethanologenic yeast by metabolic engineering or synthetic biology.

show abstract

“…The presumably zinc-containing L-threonine dehydrogenase from T. kodakaraensis was highly stable, and this dehydrogenase had a half-life at 85°C of more than 2 h (5). In the family of zinc-containing ADHs, those from the hyperthermophilic archaea T. kodakaraensis, P. horikoshii, S. solfataricus, and A. pernix contain both catalytic and structural zincs and have been extensively studied in terms of structure, catalysis, function, or regulation (2,16,19,(25)(26)(27)(28)52). However, no ADHs containing only catalytic zinc have been characterized for Thermococcus species yet.…”

mentioning

confidence: 99%

Characterization of a Zinc-Containing Alcohol Dehydrogenase with Stereoselectivity from the Hyperthermophilic Archaeon Thermococcus guaymasensis

Ying

2011

J Bacteriol

View full text Add to dashboard Cite

An alcohol dehydrogenase (ADH) from hyperthermophilic archaeon Thermococcus guaymasensis was purified to homogeneity and was found to be a homotetramer with a subunit size of 40 ؎ 1 kDa. The gene encoding the enzyme was cloned and sequenced; this gene had 1,095 bp, corresponding to 365 amino acids, and showed high sequence homology to zinc-containing ADHs and L-threonine dehydrogenases with binding motifs of catalytic zinc and NADP ؉ . Metal analyses revealed that this NADP ؉ -dependent enzyme contained 0.9 ؎ 0.03 g-atoms of zinc per subunit. It was a primary-secondary ADH and exhibited a substrate preference for secondary alcohols and corresponding ketones. Particularly, the enzyme with unusual stereoselectivity catalyzed an anti-Prelog reduction of racemic (R/S)-acetoin to (2R,3R)-2,3-butanediol and meso-2,3-butanediol. The optimal pH values for the oxidation and formation of alcohols were 10.5 and 7.5, respectively. Besides being hyperthermostable, the enzyme activity increased as the temperature was elevated up to 95°C. The enzyme was active in the presence of methanol up to 40% (vol/vol) in the assay mixture. The reduction of ketones underwent high efficiency by coupling with excess isopropanol to regenerate NADPH. The kinetic parameters of the enzyme showed that the apparent K m values and catalytic efficiency for NADPH were 40 times lower and 5 times higher than those for NADP ؉ , respectively. The physiological roles of the enzyme were proposed to be in the formation of alcohols such as ethanol or acetoin concomitant to the NADPH oxidation.

show abstract

Translational and transcriptional analysis of Sulfolobus solfataricus P2 to provide insights into alcohol and ketone utilisation

Cited by 20 publications

References 53 publications

MarR-Like Transcriptional Regulator Involved in Detoxification of Aromatic Compounds in Sulfolobus solfataricus

MarR-Like Transcriptional Regulator Involved in Detoxification of Aromatic Compounds in Sulfolobus solfataricus

Comparative Proteomic Analysis of Tolerance and Adaptation of EthanologenicSaccharomyces cerevisiaeto Furfural, a Lignocellulosic Inhibitory Compound

Characterization of a Zinc-Containing Alcohol Dehydrogenase with Stereoselectivity from the Hyperthermophilic Archaeon Thermococcus guaymasensis

Contact Info

Product

Resources

About