Construction of brewer's yeasts secreting fungal endo-�-glucanase

Penttilä, Merja; Suihko, Maija‐Liisa; Lehtinen, Ulla; Nikkola, Matti; Knowles, Jonathan

doi:10.1007/bf00434818

Cited by 54 publications

(21 citation statements)

References 20 publications

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“…Several cellulase genes from various fungi have also been expressed in S. cerevisiae. Five endo-␤-1,4-glucanases (encoded by egl1, egl2, egl3, egl4, and egl5) and two cellobiohydrolases (CBHI and CBHII) from T. reesei were efficiently secreted into the culture medium by S. cerevisiae transformants (4,27,521,522,525,586,588,655,783,784). Similar results were observed when the endo-␤-1,4-glucanase gene from Trichoderma longibrachiatum was expressed in a wine yeast strain (528,529).…”

Section: Heterologous Cellulase Expression In Bacteriasupporting

confidence: 72%

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Lynd

Weimer

Zyl³

et al. 2002

Microbiol Mol Biol Rev

4,009

2,868

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Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial cellulose utilization, kinetics of microbial cellulose utilization, and contrasting features compared to soluble substrate kinetics. A biological perspective on processing cellulosic biomass is presented, including features of pretreated substrates and alternative process configurations. Organism development is considered for “consolidated bioprocessing” (CBP), in which the production of cellulolytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products occur in one step. Two organism development strategies for CBP are examined: (i) improve product yield and tolerance in microorganisms able to utilize cellulose, or (ii) express a heterologous system for cellulose hydrolysis and utilization in microorganisms that exhibit high product yield and tolerance. A concluding discussion identifies unresolved issues pertaining to microbial cellulose utilization, suggests approaches by which such issues might be resolved, and contrasts a microbially oriented cellulose hydrolysis paradigm to the more conventional enzymatically oriented paradigm in both fundamental and applied contexts

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Section: Heterologous Cellulase Expression In Bacteriasupporting

confidence: 72%

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Lynd

Weimer

Zyl³

et al. 2002

Microbiol Mol Biol Rev

4,009

2,868

View full text Add to dashboard Cite

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“…The co-transformation method (Penttila et al 1987b) based on selection for copper-resistant colonies (Henderson et al 1985) was used. The PGK1/egll expression cassette was released from plasmid pMN20 with HindlII and the ADH1/egI1 cassette from plasmid pMN200 with BamHI, the ends were made blunt-ended with Klenow and the fragments carrying the expression cassettes were isolated from an agarose gel.…”

Section: Methodsmentioning

confidence: 99%

“…* Present address: University of Helsinki, Department of General Microbiology, Helsinki, Finland ** Present address: Hiirlimann Brewery, ZOrich, Switzerland Offprint requests to: M.-L. Suihko Hinchliffe and Box 1985) and Trichoderma reesei (Penttila et al 1987b) have been transformed to brewer's yeast.…”

Section: Introductionmentioning

confidence: 99%

Construction and analysis of recombinant glucanolytic brewer's yeast strains

Suihko

Lehtinen

Zurbriggen

et al. 1991

Appl Microbiol Biotechnol

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Four different types of endo-fl-l,4-glucanase active bottom-fermenting brewer's yeast strains were constructed using recombinant DNA technology. To study the effects of different promoters, copy numbers and integration sites, the eoll gene of the filamentous fungus Trichoderma reesei was inserted between the promoter and terminator regions of either the PGK1 or ADH1 gene of yeast. The eoll gene was transferred to the industrial brewer's yeast on a multicopy plasmid or alternatively integrated into the LEU2, PGK1 or A D H 1 locus of the yeast. Integration into the PGK1 or A D H 1 locus did not affect the brewing properties of the yeast or the quality of the finished beer. Integration into the LEU2 locus, however, decreased the metabolic activity of yeast and prolonged fermentation was needed. In pilot brewing conditions the PGK1 promoter was stronger than that of ADH1. Even a single copy of the eoH gene in the PGK1 integrant strains gave rise to sufficient enzyme activity for the hydrolysis even of unusually high total amounts of fl-glucans in worts.

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“…The latter option serves as an obvious task for metabolic engineering whereby the substrate range is extended to include ␤-glucans, and consequently the process performance of beer production may be improved. ␤-Glucanases of Bacillus subtilis (14), Trichoderma reesei (104,105), and barley (99) have successfully been expressed in S. cerevisiae, and active enzymes were secreted. The production of ␤-glucanase did not affect beer quality, and, furthermore, the ␤-glucans were efficiently degraded, resulting in an improved filterability (105).…”

Section: Improvement Of Process Performancementioning

confidence: 99%

“…␤-Glucanases of Bacillus subtilis (14), Trichoderma reesei (104,105), and barley (99) have successfully been expressed in S. cerevisiae, and active enzymes were secreted. The production of ␤-glucanase did not affect beer quality, and, furthermore, the ␤-glucans were efficiently degraded, resulting in an improved filterability (105). An improved process performance is often accomplished in association with other aims, such as was demonstrated in the last example, where an extended substrate range also was obtained.…”

Section: Improvement Of Process Performancementioning

confidence: 99%

Metabolic Engineering of Saccharomyces cerevisiae

Østergaard

Olsson

Nielsen

2000

Microbiol Mol Biol Rev

405

213

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SUMMARY Comprehensive knowledge regarding Saccharomyces cerevisiae has accumulated over time, and today S. cerevisiae serves as a widley used biotechnological production organism as well as a eukaryotic model system. The high transformation efficiency, in addition to the availability of the complete yeast genome sequence, has facilitated genetic manipulation of this microorganism, and new approaches are constantly being taken to metabolicially engineer this organism in order to suit specific needs. In this paper, strategies and concepts for metabolic engineering are discussed and several examples based upon selected studies involving S. cerevisiae are reviewed. The many different studies of metabolic engineering using this organism illustrate all the categories of this multidisciplinary field: extension of substrate range, improvements of producitivity and yield, elimination of byproduct formation, improvement of process performance, improvements of cellular properties, and extension of product range including heterologous protein production.

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Construction of brewer's yeasts secreting fungal endo-�-glucanase

Cited by 54 publications

References 20 publications

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Construction and analysis of recombinant glucanolytic brewer's yeast strains

Metabolic Engineering of Saccharomyces cerevisiae

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