Aerobic growth physiology of Saccharomyces cerevisiae on sucrose is strain-dependent

AbstractPresent knowledge on the quantitative aerobic physiology of the yeast Saccharomyces cerevisiae during growth on sucrose as sole carbon and energy source is limited to either adapted cells or to the model laboratory strain CEN.PK113-7D. To broaden our understanding of this matter and open novel opportunities for sucrose-based biotechnological processes, we characterized three strains, with distinct backgrounds, during aerobic batch bioreactor cultivations. Our results reveal that sucrose metabolism in S. cerevisiae is a strain-specific trait. Each strain displayed a distinct extracellular hexose concentration and invertase activity profiles. Especially, the inferior maximum specific growth rate (0.21 h−1) of the CEN.PK113-7D strain, with respect to that of strains UFMG-CM-Y259 (0.37 h−1) and JP1 (0.32 h−1), could be associated to its low invertase activity (0.04 to 0.09 U mgDM−1). Moreover, comparative experiments with glucose or fructose alone, or in combination, suggest mixed mechanisms of sucrose utilization by the industrial strain JP1, and points out the remarkable ability of the wild isolate UFMG-CM-259 to grow faster on sucrose than on glucose in a well-controlled cultivation system. This work hints to a series of metabolic traits that can be exploited to increase sucrose catabolic rates and bioprocess efficiency.Abstract Figure

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Aerobic growth physiology of Saccharomyces cerevisiae on sucrose is strain-dependent

FEMS Yeast Research ◽

10.1093/femsyr/foab021 ◽

2021 ◽

Vol 21 (3) ◽

Author(s):

Carla Inês Soares Rodrigues ◽

Aljoscha Wahl ◽

Andreas K Gombert

Keyword(s):

Saccharomyces Cerevisiae ◽

Laboratory Strain ◽

Invertase Activity ◽

Yeast Saccharomyces Cerevisiae ◽

Wild Isolate ◽

Growth Physiology ◽

Batch Bioreactor ◽

Specific Trait ◽

Biotechnological Processes ◽

Sucrose Utilization

ABSTRACT Present knowledge on the quantitative aerobic physiology of the yeast Saccharomyces cerevisiae during growth on sucrose as sole carbon and energy source is limited to either adapted cells or to the model laboratory strain CEN.PK113-7D. To broaden our understanding of this matter and open novel opportunities for sucrose-based biotechnological processes, we characterized three strains, with distinct backgrounds, during aerobic batch bioreactor cultivations. Our results reveal that sucrose metabolism in S. cerevisiae is a strain-specific trait. Each strain displayed distinct extracellular hexose concentrations and invertase activity profiles. Especially, the inferior maximum specific growth rate (0.21 h-1) of the CEN.PK113-7D strain, with respect to that of strains UFMG-CM-Y259 (0.37 h-1) and JP1 (0.32 h-1), could be associated to its low invertase activity (0.04–0.09 U/mgDM). Moreover, comparative experiments with glucose or fructose alone, or in combination, suggest mixed mechanisms of sucrose utilization by the industrial strain JP1, and points out the remarkable ability of the wild isolate UFMG-CM-259 to grow faster on sucrose than on glucose in a well-controlled cultivation system. This work hints to a series of metabolic traits that can be exploited to increase sucrose catabolic rates and bioprocess efficiency.

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A cytofluorimetric analysis of a Saccharomyces cerevisiae population cultured in a fed-batch bioreactor

PLoS ONE ◽

10.1371/journal.pone.0248382 ◽

2021 ◽

Vol 16 (6) ◽

pp. e0248382

Author(s):

Emanuela Palomba ◽

Valentina Tirelli ◽

Elisabetta de Alteriis ◽

Palma Parascandola ◽

Carmine Landi ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Batch Culture ◽

Time Course ◽

Reference Model ◽

Product Yield ◽

Fed Batch ◽

Batch Bioreactor ◽

Biotechnological Processes ◽

Fed Batch Culture

The yeast Saccharomyces cerevisiae is a reference model system and one of the widely used microorganisms in many biotechnological processes. In industrial yeast applications, combined strategies aim to maximize biomass/product yield, with the fed-batch culture being one of the most frequently used. Flow cytometry (FCM) is widely applied in biotechnological processes and represents a key methodology to monitor cell population dynamics. We propose here an application of FCM in the analysis of yeast cell cycle along the time course of a typical S. cerevisiae fed-batch culture. We used two different dyes, SYTOX Green and SYBR Green, with the aim to better define each stage of cell cycle during S. cerevisiae fed-batch culture. The results provide novel insights in the use of FCM cell cycle analysis for the real-time monitoring of S. cerevisiae bioprocesses.

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Faculty Opinions recommendation of Elevated evolutionary rates in the laboratory strain of Saccharomyces cerevisiae.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1023635.280653 ◽

2005 ◽

Author(s):

Karin Schmitt

Keyword(s):

Saccharomyces Cerevisiae ◽

Laboratory Strain ◽

Evolutionary Rates

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Expression of dehydrin gene from Arctic Cerastium arcticum increases abiotic stress tolerance and enhances the fermentation capacity of a genetically engineered Saccharomyces cerevisiae laboratory strain

Applied Microbiology and Biotechnology ◽

10.1007/s00253-013-4729-9 ◽

2013 ◽

Vol 97 (20) ◽

pp. 8997-9009 ◽

Cited By ~ 12

Author(s):

Il-Sup Kim ◽

Hyun-Young Kim ◽

Young-Saeng Kim ◽

Han-Gu Choi ◽

Sung-Ho Kang ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Abiotic Stress ◽

Stress Tolerance ◽

Abiotic Stress Tolerance ◽

Laboratory Strain ◽

Genetically Engineered ◽

Fermentation Capacity ◽

Engineered Saccharomyces Cerevisiae ◽

Dehydrin Gene

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Stimulatory effects of 2,4-dichlorophenoxyacetic acid and of 1-naphthylacetic acid on sucrose level, invertase activity and sucrose utilization in the latex ofHevea brasiliensis

Planta ◽

10.1007/bf01391120 ◽

1969 ◽

Vol 88 (2) ◽

pp. 144-153 ◽

Cited By ~ 20

Author(s):

Jaroslav Tup�

Keyword(s):

Invertase Activity ◽

Dichlorophenoxyacetic Acid ◽

Naphthylacetic Acid ◽

2,4 Dichlorophenoxyacetic Acid ◽

Sucrose Utilization ◽

Sucrose Level

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Comparison of the Glycolytic and Alcoholic Fermentation Pathways of Hanseniaspora vineae with Saccharomyces cerevisiae Wine Yeasts

Fermentation ◽

10.3390/fermentation6030078 ◽

2020 ◽

Vol 6 (3) ◽

pp. 78 ◽

Cited By ~ 1

Author(s):

María José Valera ◽

Eduardo Boido ◽

Eduardo Dellacassa ◽

Francisco Carrau

Keyword(s):

Saccharomyces Cerevisiae ◽

Alcoholic Fermentation ◽

Invertase Activity ◽

Wine Industry ◽

Wine Yeasts ◽

Spontaneous Fermentation ◽

Fermentative Capacity ◽

Definition Of ◽

Concept Framework ◽

Fast Evolution

Hanseniaspora species can be isolated from grapes and grape musts, but after the initiation of spontaneous fermentation, they are displaced by Saccharomyces cerevisiae. Hanseniaspora vineae is particularly valuable since this species improves the flavour of wines and has an increased capacity to ferment relative to other apiculate yeasts. Genomic, transcriptomic, and metabolomic studies in H. vineae have enhanced our understanding of its potential utility within the wine industry. Here, we compared gene sequences of 12 glycolytic and fermentation pathway enzymes from five sequenced Hanseniaspora species and S. cerevisiae with the corresponding enzymes encoded within the two sequenced H. vineae genomes. Increased levels of protein similarity were observed for enzymes of H. vineae and S. cerevisiae, relative to the remaining Hanseniaspora species. Key differences between H. vineae and H. uvarum pyruvate kinase enzymes might explain observed differences in fermentative capacity. Further, the presence of eight putative alcohol dehydrogenases, invertase activity, and sulfite tolerance are distinctive characteristics of H. vineae, compared to other Hanseniaspora species. The definition of two clear technological groups within the Hanseniaspora genus is discussed within the slow and fast evolution concept framework previously discovered in these apiculate yeasts.

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Association of Constitutive Hyperphosphorylation of Hsf1p with a Defective Ethanol Stress Response in Saccharomyces cerevisiae Sake Yeast Strains

Applied and Environmental Microbiology ◽

10.1128/aem.06341-11 ◽

2011 ◽

Vol 78 (2) ◽

pp. 385-392 ◽

Cited By ~ 18

Author(s):

Chiemi Noguchi ◽

Daisuke Watanabe ◽

Yan Zhou ◽

Takeshi Akao ◽

Hitoshi Shimoi

Keyword(s):

Saccharomyces Cerevisiae ◽

Stress Response ◽

Heat Shock ◽

Laboratory Strain ◽

Underlying Mechanism ◽

Ethanol Stress ◽

Heat Shock Element ◽

Content Type ◽

Yeast Strains ◽

Acute Ethanol

ABSTRACTModern sake yeast strains, which produce high concentrations of ethanol, are unexpectedly sensitive to environmental stress during sake brewing. To reveal the underlying mechanism, we investigated a well-characterized yeast stress response mediated by a heat shock element (HSE) and heat shock transcription factor Hsf1p inSaccharomyces cerevisiaesake yeast. The HSE-lacZactivity of sake yeast during sake fermentation and under acute ethanol stress was severely impaired compared to that of laboratory yeast. Moreover, the Hsf1p of modern sake yeast was highly and constitutively hyperphosphorylated, irrespective of the extracellular stress. SinceHSF1allele replacement did not significantly affect the HSE-mediated ethanol stress response or Hsf1p phosphorylation patterns in either sake or laboratory yeast, the regulatory machinery of Hsf1p is presumed to function differently between these types of yeast. To identify phosphatases whose loss affected the control of Hsf1p, we screened a series of phosphatase gene deletion mutants in a laboratory strain background. Among the 29 mutants, a Δppt1mutant exhibited constitutive hyperphosphorylation of Hsf1p, similarly to the modern sake yeast strains, which lack the entirePPT1gene locus. We confirmed that the expression of laboratory yeast-derived functionalPPT1recovered the HSE-mediated stress response of sake yeast. In addition, deletion ofPPT1in laboratory yeast resulted in enhanced fermentation ability. Taken together, these data demonstrate that hyperphosphorylation of Hsf1p caused by loss of thePPT1gene at least partly accounts for the defective stress response and high ethanol productivity of modern sake yeast strains.

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Exploiting the Diversity of Saccharomycotina Yeasts To Engineer Biotin-Independent Growth of Saccharomyces cerevisiae

Applied and Environmental Microbiology ◽

10.1128/aem.00270-20 ◽

2020 ◽

Vol 86 (12) ◽

Cited By ~ 1

Author(s):

Anna K. Wronska ◽

Meinske P. Haak ◽

Ellen Geraats ◽

Eva Bruins Slot ◽

Marcel van den Broek ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Large Scale ◽

De Novo ◽

Laboratory Strain ◽

Optimal Growth ◽

Industrial Applications ◽

Fast Growth ◽

Growth Media ◽

Free Medium ◽

Content Type

ABSTRACT Biotin, an important cofactor for carboxylases, is essential for all kingdoms of life. Since native biotin synthesis does not always suffice for fast growth and product formation, microbial cultivation in research and industry often requires supplementation of biotin. De novo biotin biosynthesis in yeasts is not fully understood, which hinders attempts to optimize the pathway in these industrially relevant microorganisms. Previous work based on laboratory evolution of Saccharomyces cerevisiae for biotin prototrophy identified Bio1, whose catalytic function remains unresolved, as a bottleneck in biotin synthesis. This study aimed at eliminating this bottleneck in the S. cerevisiae laboratory strain CEN.PK113-7D. A screening of 35 Saccharomycotina yeasts identified six species that grew fast without biotin supplementation. Overexpression of the S. cerevisiae BIO1 (ScBIO1) ortholog isolated from one of these biotin prototrophs, Cyberlindnera fabianii, enabled fast growth of strain CEN.PK113-7D in biotin-free medium. Similar results were obtained by single overexpression of C. fabianii BIO1 (CfBIO1) in other laboratory and industrial S. cerevisiae strains. However, biotin prototrophy was restricted to aerobic conditions, probably reflecting the involvement of oxygen in the reaction catalyzed by the putative oxidoreductase CfBio1. In aerobic cultures on biotin-free medium, S. cerevisiae strains expressing CfBio1 showed a decreased susceptibility to contamination by biotin-auxotrophic S. cerevisiae. This study illustrates how the vast Saccharomycotina genomic resources may be used to improve physiological characteristics of industrially relevant S. cerevisiae. IMPORTANCE The reported metabolic engineering strategy to enable optimal growth in the absence of biotin is of direct relevance for large-scale industrial applications of S. cerevisiae. Important benefits of biotin prototrophy include cost reduction during the preparation of chemically defined industrial growth media as well as a lower susceptibility of biotin-prototrophic strains to contamination by auxotrophic microorganisms. The observed oxygen dependency of biotin synthesis by the engineered strains is relevant for further studies on the elucidation of fungal biotin biosynthesis pathways.

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