Expression of the alpha3/beta1 isoform of human Na,K-ATPase in the methylotrophic yeast Pichia pastoris.

Abstract:

:Na,K-ATPase is a crucial enzyme for ion homeostasis in human tissues. Different isozymes are produced by assembly of four alpha- and three beta-subunits. The expression of the alpha3/beta1 isozyme is confined to brain and heart. Its heterologous production has so far never been attempted in a lower eukaryote. In this work we explored whether the methylotrophic yeast Pichia pastoris is capable of expressing the alpha3/beta1 isoform of human Na,K-ATPase. cDNAs encoding the alpha(3) and the beta(1)-subunits were cloned under the control of the inducible promoter of Pichia pastoris alcohol oxidase 1. Pichia pastoris could express the single alpha3- and beta1-subunits and even coexpress them after methanol induction. beta1-subunit was produced as a major 44-kDa glycosylated polypeptide and alpha3 as a 110-kDa unglycosylated polypeptide. Expression at the plasma membrane was limited in shaking flask cultures but by cultivating P. pastoris cells in a fermenter there was a 10-fold increase of the number of ouabain binding sites per cell. The exported enzyme was estimated to be about 0.230 mg L(-1) at the end of a bioreactor run. Na,K-ATPase proved active and the dissociation constant of the recombinant enzyme-ouabain interaction was determined.

journal_name

FEMS Yeast Res

journal_title

FEMS yeast research

authors

Reina C,Padoani G,Carotti C,Merico A,Tripodi G,Ferrari P,Popolo L

doi

10.1111/j.1567-1364.2007.00227.x

subject

Has Abstract

pub_date

2007-06-01 00:00:00

pages

585-94

issue

4

eissn

1567-1356

issn

1567-1364

pii

FYR227

journal_volume

7

pub_type

杂志文章
  • Vaccines based on whole recombinant Saccharomyces cerevisiae cells.

    abstract::The ultimate goal of therapeutic vaccines is to activate and exploit the patient's own immune system to vigorously and dynamically seek and eradicate established malignant or virally infected cells. Therapeutic vaccines also offer the potential for preventing disease recurrence. Saccharomyces cerevisiae-based vaccines...

    journal_title:FEMS yeast research

    pub_type: 杂志文章,评审

    doi:10.1111/j.1567-1364.2010.00665.x

    authors: Ardiani A,Higgins JP,Hodge JW

    更新日期:2010-12-01 00:00:00

  • Sequence modification of the master regulator Pdr1 interferes with its transcriptional autoregulation and confers altered azole resistance in Candida glabrata.

    abstract::The transcriptional regulator Pdr1 plays a positive role in regulating azole drug resistance in Candida glabrata. Previous studies have shown the importance of the carboxyl (C)-terminal sequence of Pdr1 in fulfilling its function, as this region mediates interactions between Pdr1 and the co-activator Gal11A and is cru...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1093/femsyr/foy038

    authors: Tian Y,Gao N,Ni Q,Mao Y,Dong D,Huang X,Jiang C,Li Z,Zhang L,Wang X,Peng Y,Chen C

    更新日期:2018-06-01 00:00:00

  • The oxygen level determines the fermentation pattern in Kluyveromyces lactis.

    abstract::Yeasts belonging to the lineage that underwent whole-genome duplication (WGD) possess a good fermentative potential and can proliferate in the absence of oxygen. In this study, we analyzed the pre-WGD yeast Kluyveromyces lactis and its ability to grow under oxygen-limited conditions. Under these conditions, K. lactis ...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2009.00528.x

    authors: Merico A,Galafassi S,Piskur J,Compagno C

    更新日期:2009-08-01 00:00:00

  • Characteristics of Flo11-dependent flocculation in Saccharomyces cerevisiae.

    abstract::The FLO11-encoded flocculin is required for a variety of important phenotypes in Saccharomyces cerevisiae, including flocculation, adhesion to agar and plastic, invasive growth, pseudohyphae formation and biofilm development. We present evidence that Flo11p belongs to the Flo1-type class of flocculins rather than to t...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1016/j.femsyr.2005.05.004

    authors: Bayly JC,Douglas LM,Pretorius IS,Bauer FF,Dranginis AM

    更新日期:2005-12-01 00:00:00

  • Genome-wide expression analyses: Metabolic adaptation of Saccharomyces cerevisiae to high sugar stress.

    abstract::The transcriptional response of laboratory strains of Saccharomyces cerevisiae to salt or sorbitol stress has been well studied. These studies have yielded valuable data on how the yeast adapts to these stress conditions. However, S. cerevisiae is a saccharophilic fungus and in its natural environment this yeast encou...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1016/S1567-1356(02)00203-9

    authors: Erasmus DJ,van der Merwe GK,van Vuuren HJ

    更新日期:2003-06-01 00:00:00

  • Cell density-dependent linoleic acid toxicity to Saccharomyces cerevisiae.

    abstract::Since the discovery of the apoptotic pathway in Saccharomyces cerevisiae, several compounds have been shown to cause apoptosis in this organism. While the toxicity of polyunsaturated fatty acids (PUFA) peroxides towards S. cerevisiae has been known for a long time, studies on the effect of nonoxidized PUFA are scarce....

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2011.00729.x

    authors: Ferreira TC,de Moraes LM,Campos EG

    更新日期:2011-08-01 00:00:00

  • Investigating the multibudded and binucleate phenotype of the yeast Zygosaccharomyces bailii growing on minimal medium.

    abstract::The yeast Zygosaccharomyces bailii, known to have peculiar resistance to several environmental constraints, is very little known with respect to its genetics and life cycle. In addition to molecular and biochemical studies, cytofluorimetric and morphological analyses can also add information necessary to shed light on...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2008.00417.x

    authors: Dato L,Sauer M,Passolunghi S,Porro D,Branduardi P

    更新日期:2008-09-01 00:00:00

  • Ume6p is required for germination and early colony development of yeast ascospores.

    abstract::Ume6p is a nonessential transcription factor that represses meiotic gene expression during vegetative growth in budding yeast. To relieve this repression, Ume6p is destroyed as cells enter meiosis and is not resynthesized until spore wall assembly. The present study reveals that spores derived from a ume6 null homozyg...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2010.00696.x

    authors: Strich R,Khakhina S,Mallory MJ

    更新日期:2011-02-01 00:00:00

  • The Saccharomyces cerevisiae Arf3 protein is involved in actin cable and cortical patch formation.

    abstract::We show that Arf3p, a member of the ADP ribosylation family, is involved in the organization of actin cables and cortical patches in Saccharomyces cerevisiae. Profilin-deficient cells (pfy1Delta) have severe growth defects and lack actin cables. Overexpression of ARF3 restores actin cables and corrects growth defects ...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2007.00239.x

    authors: Lambert AA,Perron MP,Lavoie E,Pallotta D

    更新日期:2007-09-01 00:00:00

  • Phosphorylation of Bni4 by MAP kinases contributes to septum assembly during yeast cytokinesis.

    abstract::Previous work has shown that the synthetic lethality of the slt2Δrim101Δ mutant results from a combination of factors, including improper functioning of the septum assembly machinery. Here, we identify new multicopy suppressors of this lethality including Kss1, Pcl1 and Sph1, none of which seems to be linked to the up...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1093/femsyr/fow060

    authors: Pérez J,Arcones I,Gómez A,Casquero V,Roncero C

    更新日期:2016-09-01 00:00:00

  • Tipping the balance both ways: drug resistance and virulence in Candida glabrata.

    abstract::Among existing fungal pathogens, Candida glabrata is outstanding in its capacity to rapidly develop resistance to currently used antifungal agents. Resistance to the class of azoles, which are still widely used agents, varies in proportion (from 5 to 20%) depending on geographical area. Moreover, resistance to the cla...

    journal_title:FEMS yeast research

    pub_type: 杂志文章,评审

    doi:10.1093/femsyr/fov025

    authors: Vale-Silva LA,Sanglard D

    更新日期:2015-06-01 00:00:00

  • Characterization of a biofilm-like extracellular matrix in FLO1-expressing Saccharomyces cerevisiae cells.

    abstract::Like bacteria, fungi growing in biofilms are often embedded in a so-called extracellular matrix (ECM), a complex and species-specific mixture of compounds secreted by cells in the biofilm. The precise physiological role of this ECM and its importance for the stress and drug resistance that is so characteristic of biof...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2009.00482.x

    authors: Beauvais A,Loussert C,Prevost MC,Verstrepen K,Latgé JP

    更新日期:2009-05-01 00:00:00

  • The F1-ATPase inhibitor Inh1 (IF1) affects suppression of mtDNA loss-lethality in Kluyveromyces lactis.

    abstract::Loss of mtDNA by the petite-negative yeast Kluyveromyces lactis is lethal (rho(o)-lethality). However, mutations in the alpha, beta and gamma subunits of F(1)-ATPase can suppress lethality by increasing intramitochondrial hydrolysis of ATP. Increased hydrolysis of ATP can also occur on inactivation of Inh1, the natura...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2006.00201.x

    authors: Clark-Walker GD

    更新日期:2007-08-01 00:00:00

  • Formic acid induces Yca1p-independent apoptosis-like cell death in the yeast Saccharomyces cerevisiae.

    abstract::Formic acid disrupts mitochondrial electron transport and sequentially causes cell death in mammalian ocular cells by an unidentified molecular mechanism. Here, we show that a low concentration of formic acid induces apoptosis-like cell death in the budding yeast Saccharomyces cerevisiae, with several morphological an...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2008.00375.x

    authors: Du L,Su Y,Sun D,Zhu W,Wang J,Zhuang X,Zhou S,Lu Y

    更新日期:2008-06-01 00:00:00

  • Yarrowia lipolytica: a model yeast for citric acid production.

    abstract::Every year more than 2 million tons of citric acid (CA) are produced around the world for industrial uses. Although initially extracted from citrus, the low profitability of the process and the increasing demand soon stimulated the search for more efficient methods to produce CA. Currently, most world CA demand (99%) ...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1093/femsyr/fox084

    authors: Cavallo E,Charreau H,Cerrutti P,Foresti ML

    更新日期:2017-12-01 00:00:00

  • 13C-Labeled metabolic flux analysis of a fed-batch culture of elutriated Saccharomyces cerevisiae.

    abstract::This study addresses the question of whether observable changes in fluxes in the primary carbon metabolism of Saccharomyces cerevisiae occur between the different phases of the cell division cycle. To detect such changes by metabolic flux analysis, a 13C-labeling experiment was performed with a fed-batch culture inocu...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2006.00199.x

    authors: Costenoble R,Müller D,Barl T,van Gulik WM,van Winden WA,Reuss M,Heijnen JJ

    更新日期:2007-06-01 00:00:00

  • Comparative genotyping of the Saccharomyces cerevisiae laboratory strains S288C and CEN.PK113-7D using oligonucleotide microarrays.

    abstract::To analyse the reliability and accuracy of genotype analysis with high-density oligonucleotide microarrays, this method and other experimental approaches were used to analyse genomic DNA of two popular Saccharomyces cerevisiae laboratory strains. S288C was used for systematic sequencing of 'the' S. cerevisiae genome; ...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1016/S1567-1356(03)00156-9

    authors: Daran-Lapujade P,Daran JM,Kötter P,Petit T,Piper MD,Pronk JT

    更新日期:2003-12-01 00:00:00

  • Neither 1G nor 2G fuel ethanol: setting the ground for a sugarcane-based biorefinery using an iSUCCELL yeast platform.

    abstract::First-generation (1G) fuel ethanol production in sugarcane-based biorefineries is an established economic enterprise in Brazil. Second-generation (2G) fuel ethanol from lignocellulosic materials, though extensively investigated, is currently facing severe difficulties to become economically viable. Some of the challen...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1093/femsyr/foaa027

    authors: Bermejo PM,Raghavendran V,Gombert AK

    更新日期:2020-06-01 00:00:00

  • Experimental evolution with yeast.

    abstract::Many of the difficulties of studying evolution in action can be surmounted using populations of microorganisms, such as yeast. A readily manipulated sexual system and an increasingly sophisticated array of molecular and genomic tools uniquely qualify Saccharomyces cerevisiae as an experimental subject. This minireview...

    journal_title:FEMS yeast research

    pub_type: 杂志文章,评审

    doi:10.1111/j.1567-1364.2006.00061.x

    authors: Zeyl C

    更新日期:2006-08-01 00:00:00

  • Cbc2p, Upf3p and eIF4G are components of the DRN (Degradation of mRNA in the Nucleus) in Saccharomyces cerevisiae.

    abstract::Messenger RNAs retained in the nucleus of Saccharomyces cerevisiae are subjected to a degradation system designated DRN (Degradation of mRNA in the Nucleus) that is dependent on the nuclear mRNA cap-binding protein, Cbc1p, as well as nuclear exosome component Rrp6p, a 3' to 5' exoribonuclease. DRN has been shown to ac...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/1567-1364.12180

    authors: Das S,Saha U,Das B

    更新日期:2014-09-01 00:00:00

  • Effects of inactivation of the PPN1 gene on exopolyphosphatases, inorganic polyphosphates and function of mitochondria in the yeast Saccharomyces cerevisiae.

    abstract::Mutants of Saccharomyces cerevisiae with inactivated endopolyphosphatase gene PPN1 did not grow on lactate and ethanol, and stopped growth on glucose earlier than the parent strain. Their mitochondria were defective in respiration functions and in metabolism of inorganic polyphosphates. The PPN1 mutants lacked exopoly...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1016/j.femsyr.2005.03.002

    authors: Pestov NA,Kulakovskaya TV,Kulaev IS

    更新日期:2005-06-01 00:00:00

  • Disruption of the MNN10 gene enhances protein secretion in Kluyveromyces lactis and Saccharomyces cerevisiae.

    abstract::Screening for genes affecting super-secreting phenotype of the over-secreting mutant of Kluyveromyces lactis resulted in isolation of the gene named KlMNN10, sharing high homology with Saccharomyces cerevisiae MNN10. The disruption of the KlMNN10 in Kluyveromyces lactis, as well as of MNN10 and MNN11 in Saccharomyces ...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1016/j.femsyr.2004.03.001

    authors: Bartkeviciūte D,Sasnauskas K

    更新日期:2004-09-01 00:00:00

  • Dynamics of yeast populations recovered from decaying leaves in a nonpolluted stream: a 2-year study on the effects of leaf litter type and decomposition time.

    abstract::Here we report on the results of a survey of the yeast populations occurring on submerged leaves (alder, eucalyptus and oak) in a natural mountain stream, during different phases of their decomposition and through two consecutive years. Leaf litter mass loss, total yeast counts, Shannon-Weiner index (H'), yeast commun...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2007.00218.x

    authors: Sampaio A,Sampaio JP,Leão C

    更新日期:2007-06-01 00:00:00

  • Evidence for the attachment of Hsp150/Pir2 to the cell wall of Saccharomyces cerevisiae through disulfide bridges.

    abstract::Here we present evidence that Hsp150/Pir2, a member of the Pir family of cell wall proteins, can be extracted from the purified cell walls of Saccharomyces cerevisiae by treatment with beta-mercaptoethanol, demonstrating that at least part of this protein is attached to the cell wall through disulfide bridges. We also...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2001.tb00040.x

    authors: Moukadiri I,Zueco J

    更新日期:2001-12-01 00:00:00

  • The interplay between sulphur and selenium metabolism influences the intracellular redox balance in Saccharomyces cerevisiae.

    abstract::Selenium (Se) is an essential element for most eukaryotic organisms, including humans. The balance between Se toxicity and its beneficial effects is very delicate. It has been demonstrated that a diet enriched with Se has cancer prevention potential in humans. The most popular commercial Se supplementation is selenize...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2011.00757.x

    authors: Mapelli V,Hillestrøm PR,Patil K,Larsen EH,Olsson L

    更新日期:2012-02-01 00:00:00

  • Life without putrescine: disruption of the gene-encoding polyamine oxidase in Ustilago maydis odc mutants.

    abstract::In previous communications the essential role of spermidine in Ustilago maydis was demonstrated by means of the disruption of the genes encoding ornithine decarboxylase (ODC) and spermidine synthase (SPE). However, the assignation of specific roles to each polyamine in different cellular functions was not possible bec...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2010.00675.x

    authors: Valdés-Santiago L,Guzmán-de-Peña D,Ruiz-Herrera J

    更新日期:2010-11-01 00:00:00

  • Phenotypic evaluation and characterization of 21 industrial Saccharomyces cerevisiae yeast strains.

    abstract::Microorganisms have been studied and used extensively to produce value-added fuels and chemicals. Yeasts, specifically Saccharomyces cerevisiae, receive industrial attention because of their well-known ability to ferment glucose and produce ethanol. Thousands of natural or genetically modified S. cerevisiae have been ...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1093/femsyr/foy001

    authors: Kong II,Turner TL,Kim H,Kim SR,Jin YS

    更新日期:2018-02-01 00:00:00

  • Cell length growth in fission yeast: an analysis of its bilinear character and the nature of its rate change transition.

    abstract::During their mitotic cycle, cylindrical fission yeast cells grow exclusively at their tips. Length growth starts at birth and halts at mitotic onset when the cells begin to prepare for division. While the growth pattern was initially considered to be exponential, during the last three decades an increasing amount of e...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/1567-1364.12064

    authors: Horváth A,Rácz-Mónus A,Buchwald P,Sveiczer Á

    更新日期:2013-11-01 00:00:00

  • Pkc1 and actin polymerisation activities play a role in ribosomal gene repression associated with secretion impairment caused by oxidative stress.

    abstract::In Saccharomyces cerevisiae, the cell integrity pathway plays a role in the oxidative stress response. In this study, we show that the Pkc1 protein mediates oxidative signalling by helping to downregulate ribosomal gene expression when cells are exposed to hydrogen peroxide. An active actin cytoskeleton is required fo...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1111/j.1567-1364.2011.00754.x

    authors: Mitjana FV,Petkova MI,Pujol-Carrion N,de la Torre-Ruiz MA

    更新日期:2011-12-01 00:00:00

  • Elucidating cellular mechanisms of Saccharomyces cerevisiae tolerant to combined lignocellulosic-derived inhibitors using high-throughput phenotyping and multiomics analyses.

    abstract::A robust cell factory that can tolerate combined inhibitory lignocellulosic compounds is essential for the cost-effective lignocellulose-based production of second-generation bioethanol and other bulk chemicals. Following high-throughput phenotyping of a yeast genomic overexpression library, we identified a Saccharomy...

    journal_title:FEMS yeast research

    pub_type: 杂志文章

    doi:10.1093/femsyr/foy106

    authors: Unrean P,Gätgens J,Klein B,Noack S,Champreda V

    更新日期:2018-12-01 00:00:00