Processing and secretion of barley (1–3, 1–4)-β-glucanase in yeast

Olsen, Ole; Thomsen, Kristine Rømer

doi:10.1007/bf02907583

Cited by 13 publications

(4 citation statements)

References 34 publications

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“…With the aim of constructing a glucanolytic brewing yeast strain, the ␤-1,3-1,4-glucanase gene from barley was cloned and expressed in S. cerevisiae (308). This gene was fused in frame to the signal sequences of the mouse ␣-amylase, yeast PHO5-encoded phosphatase, and SUC2-encoded invertase (504). These constructs were inserted behind the ADH1 and PGK1 promoters.…”

Section: Heterologous Cellulase Expression In Bacteriamentioning

confidence: 99%

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Lynd

Weimer

Zyl³

et al. 2002

Microbiol Mol Biol Rev

4,014

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 Bacteriamentioning

confidence: 99%

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Lynd

Weimer

Zyl³

et al. 2002

Microbiol Mol Biol Rev

4,014

2,868

View full text Add to dashboard Cite

show abstract

“…Although the introduction of flocculence to brewer's yeast is a convenient method to separate the yeast from the brewing, beer filtration is still an important separation technique in the brewing industry. Endo-l,3-1,4-β-glucanases from B. subtilis (Cantwell et al, 1986), Trichoderma reesei (Panttilä et al, 1987a;1987b), and barley (Olsen and Thomsen, 1989) have successfully been expressed in S. cerevisiae, and active enzymes were secreted. The production of β-glucanase did not affect beer quality, and furthermore, the β-glucans were efficiently degraded, resulting in an improved filterability (Panttilä et al, 1987a).…”

Section: Introductionmentioning

confidence: 99%

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

Zhang

Chen

et al. 2008

J. Zhejiang Univ. Sci. B

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The bglS gene encoding endo-l,3-1,4-beta-glucanase from Bacillus subtilis was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone alpha-factor (MFalpha1(S)), and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction (PCR)-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae (SC-betaG) was preliminarily screened by the clearing hydrolysis zone formed after the barley beta-glucan was hydrolyzed in the plate and no proteinase A (PrA) activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-l,3-1,4-beta-glucanase assay methods showed that the recombinant strain SC-betaG had high endo-l,3-1,4-beta-glucanase expression level with the maximum of 69.3 U/(h.ml) after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-l,3-1,4-beta-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.

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“…Transgenic yeasts synthesize barley 0-glucanase and secrete this enzyme into the culture medium. Five milligrams of enzyme per liter could be produced (27). Numerous site-directed mutations, which lead to exchanges of individual amino acids of the barley enzyme, were made and tested for better thermostability but without success.…”

Section: Intragenic Heterosis In Engineering An Improved Malt Enzymementioning

confidence: 99%

Genetic engineering and plant breeding, especially cereals

Wettstein

1993

Food Reviews International

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Over the last 5000 years cereals have been bred for food, feed, and beverages by selection of spontaneous mutations and random hybrids. Since the turn of the century, crosses with defined parents, and since 1927 artificially induced mutations, have been used to create variability on which selection of new varieties is based. It is pointed out that hybrid corn and transfer of rust-resistant genes from wild species into chromosomes of bread wheat was preceded by decades of basic research. Genetic transformation is an additional tool for the breeder to introduce novel genes in a rational manner and will complement but not replace the existing efficient breeding methods. Genetic transformation has been demonstrated in maize, rice, and wheat, while techniques to obtain transgenic barley plants are still being developed. Our present knowledge on the endosperm-specific expression of storage proteins and the modulation of this expression by transcriptional activators is reviewed. Breeding strategies for altered protein quality and for proanthocyanidin-free malting barley are presented. Engineering of an improved malt enzyme, a heat stable (1-3,1-4)-β-glucanase, is described. 411

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Processing and secretion of barley (1–3, 1–4)-β-glucanase in yeast

Cited by 13 publications

References 34 publications

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Microbial Cellulose Utilization: Fundamentals and Biotechnology

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

Genetic engineering and plant breeding, especially cereals

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