Comparison of the Gene Expression Patterns of Alcohol Dehydrogenase Isozymes in the Thermotolerant Yeast<i>Kluyveromyces marxianus</i>and Their Physiological Functions

Lertwattanasakul, Noppon; Sootsuwan, Kaewta; Limtong, Savitree; Thanonkeo, Pornthap; Yamada, Mamoru

doi:10.1271/bbb.60622

Cited by 28 publications

(29 citation statements)

References 46 publications

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“…3). The regulation of adhB, which has not been fully defined, seems to be unique and different from S. cerevisiae ADH2 [27] or Kluyveromyces marxianus ADH4 [28], which are repressed by glucose and induced by ethanol, respectively, and proposed to be involved in ethanol degradation. Z. mobilis AdhA appears to mainly perform ethanol synthesis [22,25].…”

Section: Discussionmentioning

confidence: 99%

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Thermotolerant Zymomonas mobilis: Comparison of Ethanol Fermentation Capability with that of an Efficient Type Strain

Sootsuwan¹,

Irie²,

Murata³

et al. 2007

TOBIOTJ

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Abstract:Zymomonas mobilis is an alternative microorganism to Saccharomyces cerevisiae for ethanol production. To find a thermotolerant Z. mobilis strain, the growth and ethanol production of four isolates in Thailand were compared with those of the efficient strain ZM4 (NRRL B-14023) at different temperatures. One of the selected strains, TISTR 405, was found to grow and produce ethanol even at 39˚C to an extent similar to that at 30˚C, and the growth and ethanol productivity at 39˚C were better than those of ZM4 at 30˚C, suggesting that TISTR 405 is suitable for ethanol fermentation at high temperatures. Analysis of genes directly related to ethanol formation or degradation, adhA, adhB and pdc, encoding alcohol dehydrogenase (Adh) A, AdhB and pyruvate decarboxylase, respectively, revealed that these genes were highly conserved in both strains. Comparison of their gene expression and activity of the products in both TISTR 405 and ZM4 at different temperatures or growth phases indicated that there was not a great difference at the transcriptional level, but the total activity of AdhA and AdhB in TISTR 405 was higher than that in ZM4. Both strains showed a significant increase in AdhB activity in the stationary phase.

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Section: Discussionmentioning

confidence: 99%

“…Z. mobilis AdhA appears to mainly perform ethanol synthesis [22,25]. The AdhA is like S. cerevisiae Adh1 [29] or K. marxianus Adh1 [28], which are constitutively expressed and are involved in ethanol formation but might have additional regulations responsible for increase in activity in the stationary phase.…”

Section: Discussionmentioning

confidence: 99%

Thermotolerant Zymomonas mobilis: Comparison of Ethanol Fermentation Capability with that of an Efficient Type Strain

Sootsuwan¹,

Irie²,

Murata³

et al. 2007

TOBIOTJ

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“…Escherichia coli strain DH5 (supE44 ÁlacU169 ('80 lacZÁM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1) was used, as described previously. 4) When required, geneticin (G418) (150 mg/ml), amplicillin (50 mg/ml), and kanamycin (25 mg/ml) were added. The glucose and ethanol concentrations were determined as described previously.…”

Section: Methodsmentioning

confidence: 99%

“…KmADH3 is expressed in the presence of respiratory carbon sources, including glycerol. 4) In contrast, the expression level of KmADH4 is high in the presence of ethanol but low in the presence of glycerol. The expression levels of both genes are very low in glucose medium.…”

Section: )mentioning

confidence: 97%

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The Crucial Role of Alcohol Dehydrogenase Adh3 inKluyveromyces marxianusMitochondrial Metabolism

Lertwattanasakul

Shigemoto

Rodrussamee

et al. 2009

Bioscience, Biotechnology, and Biochemistry

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Deletion analysis of Pichia pastoris alcohol dehydrogenase 2 (ADH2) promoter and development of synthetic promoters

Erden-Karaoğlan

Karaoglan

Yılmaz

et al. 2021

Biotechnology Journal

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Pichia pastoris (Komagataella phaffii) is a non-conventional Crabtree-negative yeast with the capability of reaching very high cell densities in a fed-batch fermentation process.The alcohol dehydrogenase (ADH) genes of P. pastoris involved in ethanol metabolism were identified and were previously characterized. This work aimed to extend current knowledge of the regulation of the ADH2 promoter. To this end, we first determined the upstream activator (UAS) and repressor (URS) sequences of the promoter by deletion assays. Two upstream activator sites have been identified, positioned between -900 and -801 bp, and -284 and -108 bp upstream of the ADH2 transcription start site. The sequences positioned between -361 and -262 bp had a negative effect on the promoter activity and designated a repressor sequence (URS). We then demonstrated that Mxr1 (methanol expression regulator 1) transcription factor activates the ADH2 promoter through the direct interaction with UAS regions in response to ethanol. Furthermore, five different synthetic promoters were constructed by adding or deleting the regulatory sites. These synthetic promoters were tested for extracellular xylanase production at shake flask level by inducing with ethanol. These promoter variants improved the xylanase production ranging between 165% and 200% of the native promoter. The synthetic promoter 5 (SNT5) that displayed the highest activity was further evaluated at the fermenter scale. The modification in the promoter features might have several implications for industrial processes where decoupling the cell growth and product formation is advantageous.

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Comparison of the Gene Expression Patterns of Alcohol Dehydrogenase Isozymes in the Thermotolerant YeastKluyveromyces marxianusand Their Physiological Functions

Cited by 28 publications

References 46 publications

Thermotolerant Zymomonas mobilis: Comparison of Ethanol Fermentation Capability with that of an Efficient Type Strain

Thermotolerant Zymomonas mobilis: Comparison of Ethanol Fermentation Capability with that of an Efficient Type Strain

The Crucial Role of Alcohol Dehydrogenase Adh3 inKluyveromyces marxianusMitochondrial Metabolism

Deletion analysis of Pichia pastoris alcohol dehydrogenase 2 (ADH2) promoter and development of synthetic promoters

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