Molecular cloning of alcohol dehydrogenase genes of the yeast Pichia stipitis and identification of the fermentative ADH

Passoth, Volkmar; Schäfer, Bernd; Liebel, Birgit; Weierstall, Thomas; Klinner, Ulrich

doi:10.1002/(sici)1097-0061(1998100)14:14<1311::aid-yea315>3.3.co;2-k

Cited by 21 publications

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“…It is possible that in D. bruxellensis, the same ADH gene is responsible for both production and consumption of ethanol, which would explain its upregulation when the glucose is almost finished. Similar observations have been made in Scheffersomyces stipitis and Wickerhamomyces anomalus [24,[31][32][33].…”

Section: Discussionsupporting

confidence: 85%

Adaptation of Dekkera bruxellensis to lignocellulose‐based substrate

Tiukova

Pita

Sundell

et al. 2014

Biotech and App Biochem

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Adaptation of Dekkera bruxellensis to lignocellulose hydrolysate was investigated. Cells of D. bruxellensis were grown for 72 and 192 H in batch and continuous culture, respectively (adapted cells). Cultivations in semisynthetic medium were run as controls (nonadapted cells). To test the adaptation, cells from these cultures were reinoculated in the lignocellulose medium, and growth and ethanol production characteristics were monitored. Cells adapted to lignocellulose hydrolysate had a shorter lag phase, grew faster, and produced a higher ethanol concentration as compared with nonadapted cells. A stability test showed that after cultivation in rich medium, cells partially lost the adapted phenotype but still showed faster growth and higher ethanol production as compared with nonadapted cells. Because alcohol dehydrogenase genes have been described to be involved in the adaptation to furfural in Saccharomyces cerevisiae, an analogous mechanism of adaptation to lignocelluloses hydrolysate of D. bruxellensis was hypothesized. However, gene expression analysis showed that genes homologous to S. cerevisiae ADH1 were not involved in the adaptation to lignocelluloses hydrolysate in D. bruxellensis.

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Section: Discussionsupporting

confidence: 85%

Adaptation of Dekkera bruxellensis to lignocellulose‐based substrate

Tiukova

Pita

Sundell

et al. 2014

Biotech and App Biochem

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“…The resultant strain VHb + had a higher specific growth rate and achieved a higher cell density under microaerobic condition. The PsADH-promoter is not only activated in P. stipitis as reported by Passoth et al (1998Passoth et al ( , 2003 and Passoth & HahnHagerdal (2000) but also in P. pastoris under low O 2 condition. The application of this VHb + strain for the high-level expression of other interested heterologous proteins in high-cell-density cultivation is currently carried out in our laboratory.…”

Section: Resultssupporting

confidence: 70%

Expression of Bacterial Hemoglobin in the Yeast, Pichia pastoris, with a Low O2-Induced Promoter

Chien

Lee

2005

Biotechnol Lett

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The PsADH2-promoter of Pichia stipitis alcohol dehydrogenase II (ADH II) gene was employed to control the expression of Vitreoscilla hemoglobin (VHb) gene in Pichia pastoris. As in P. stipitis, the promoter was also induced microaerobically in P. pastoris. The expression level of VHb in P. pastoris at low O2 tension (<5% air saturation) was 16 nmol/g dry cell wt, i.e. about 24-fold higher than that at 60% air saturation. The expressed VHb enhanced growth of P. pastoris under microaerobic conditions. The application of O2-regulated promoter in P. pastoris revealed that induction of high-level expression of heterologous protein is feasible without addition of supplementary compounds.

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“…The observed metabolic regulation patterns in response to different oxygen availability are in agreement with experimental results. Previous studies have shown an increase of alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC) activities at both a genetic and enzymatic level in S. stipitis during oxygen limitation (Cho and Jeffries 1999;Klinner et al 2005;Passoth et al 1996Passoth et al , 1998Passoth et al , 2003. Transcripts of 2-ketoglutarate dehydrogenase (KGD) and malate dehydrogenase (MDH) were decreased with decreasing oxygen level (Jeffries 2008;Skoog et al 1992).…”

Section: Discussionmentioning

confidence: 99%

Metabolic pathway analysis of Scheffersomyces (Pichia) stipitis: effect of oxygen availability on ethanol synthesis and flux distributions

Unrean

Nguyen

2012

Appl Microbiol Biotechnol

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Elementary mode analysis (EMA) identifies all possible metabolic states of the cell metabolic network. Investigation of these states can provide a detailed insight into the underlying metabolism in the cell. In this study, the flux states of Scheffersomyces (Pichia) stipitis metabolism were examined. It was shown that increasing oxygen levels led to a decrease of ethanol synthesis. This trend was confirmed by experimental evaluation of S. stipitis in glucose-xylose fermentation. The oxygen transfer rate for an optimal ethanol production was 1.8 mmol/l/h, which gave the ethanol yield of 0.40 g/g and the ethanol productivity of 0.25 g/l/h. For a better understanding of the cell's regulatory mechanism in response to oxygenation levels, EMA was used to examine metabolic flux patterns under different oxygen levels. Up- and downregulation of enzymes in the network during the change of culturing condition from oxygen limitation to oxygen sufficiency were identified. The results indicated the flexibility of S. stipitis metabolism to cope with oxygen availability. In addition, relevant genetic targets towards improved ethanol-producing strains under all oxygenation levels were identified. These targeted genes limited the metabolic functionality of the cell to function according to the most efficient ethanol synthesis pathways. The presented approach is promising and can contribute to the development of culture optimization and strain engineers for improved lignocellulosic ethanol production by S. stipitis.

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Molecular cloning of alcohol dehydrogenase genes of the yeast Pichia stipitis and identification of the fermentative ADH

Cited by 21 publications

References 0 publications

Adaptation of Dekkera bruxellensis to lignocellulose‐based substrate

Adaptation of Dekkera bruxellensis to lignocellulose‐based substrate

Expression of Bacterial Hemoglobin in the Yeast, Pichia pastoris, with a Low O2-Induced Promoter

Metabolic pathway analysis of Scheffersomyces (Pichia) stipitis: effect of oxygen availability on ethanol synthesis and flux distributions

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