Evaluation of the cellobiose-fermenting yeastBrettanomyces custersii in the simultaneous saccharification and fermentation of cellulose

Spindler, Diane D.; Wyman, Charles E.; Grohmann, K.; Philippidis, George P.

doi:10.1007/bf01021255

Cited by 59 publications

(22 citation statements)

References 12 publications

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“…␤-Glucosidases are well-known for their role in flavor development in beer and wine (74,75). Additionally, ␤-glucosidase has been shown to play a role in the fermentation of cellobiose by B. bruxellensis (76)(77)(78)(79). Interestingly, we found that ST05.12/22 did contain another ␤-glucosidase gene, which was also present in AWRI 1499 and CBS 2499.…”

Section: Discussionmentioning

confidence: 49%

Assessing Genetic Diversity among Brettanomyces Yeasts by DNA Fingerprinting and Whole-Genome Sequencing

Crauwels

Zhu

Steensels

et al. 2014

Appl Environ Microbiol

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e Brettanomyces yeasts, with the species Brettanomyces (Dekkera) bruxellensis being the most important one, are generally reported to be spoilage yeasts in the beer and wine industry due to the production of phenolic off flavors. However, B. bruxellensis is also known to be a beneficial contributor in certain fermentation processes, such as the production of certain specialty beers. Nevertheless, despite its economic importance, Brettanomyces yeasts remain poorly understood at the genetic and genomic levels. In this study, the genetic relationship between more than 50 Brettanomyces strains from all presently known species and from several sources was studied using a combination of DNA fingerprinting techniques. This revealed an intriguing correlation between the B. bruxellensis fingerprints and the respective isolation source. To further explore this relationship, we sequenced a (beneficial) beer isolate of B. bruxellensis (VIB X9085; ST05.12/22) and compared its genome sequence with the genome sequences of two wine spoilage strains (AWRI 1499 and CBS 2499). ST05.12/22 was found to be substantially different from both wine strains, especially at the level of single nucleotide polymorphisms (SNPs). In addition, there were major differences in the genome structures between the strains investigated, including the presence of large duplications and deletions. Gene content analysis revealed the presence of 20 genes which were present in both wine strains but absent in the beer strain, including many genes involved in carbon and nitrogen metabolism, and vice versa, no genes that were missing in both AWRI 1499 and CBS 2499 were found in ST05.12/22. Together, this study provides tools to discriminate Brettanomyces strains and provides a first glimpse at the genetic diversity and genome plasticity of B. bruxellensis.

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Section: Discussionmentioning

confidence: 49%

Assessing Genetic Diversity among Brettanomyces Yeasts by DNA Fingerprinting and Whole-Genome Sequencing

Crauwels

Zhu

Steensels

et al. 2014

Appl Environ Microbiol

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“…Though expensive, productivity can be improved by increasing the level of cellobiase and other cellulases (Desrochers et al, 1981;Freer and Detroy, 1983;Sternberg et al, 1977). The discovery of cellobiose-fermenting yeasts (Barnett, 1976 Freer and Detroy, 1983;Maleszka et al, 1982) represented a further improvement by eliminating the need for supplemental cellobiase (Freer, 1991;Spindler et al, 1992;Wyman et at., 1986).…”

Section: Introductionmentioning

confidence: 99%

Fermentation of Crystalline Cellulose to Ethanol by Klebsiella oxytoca Containing Chromosomally Integrated Zymomonas mobilis Genes

Doran

Ingram

1993

Biotechnology Progress

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Complete enzymatic hydrolysis of cellulose to glucose is generally required for efficient fermentation to ethanol. This hydrolysis requires endoglucanase, exoglucanase, and cellobiase. The Gram‐negative bacterium, Klebsiella oxytoca, contains the native ability to transport and metabolize cellobiose, minimizing the need for extracellular cellobiase. Strain P2 is a recombinant derivative in which the Zymomonas mobilis pdc and adhB genes have been integrated into the chromosome and expressed, directing the metabolism of pyruvate to ethanol. This organism has been evaluated in simultaneous sacchari‐fication and fermentation (SSF) experiments to determine optimal conditions and limits of performance. The temperature was varied between 32 and 40 °C over a pH range of 5.0–5.8 with 100 g/L crystalline cellulose (Sigmacell 50, Sigma Chemical Company, St. Louis, MO) as the substrate and commercial cellulase (Spezyme CE, South San Francisco, CA). A broad optimum for SSF was observed, with a pH of 5.2–5.5 and temperatures of 32–35 °C, which allowed the production of over 44 g of ethanol/L (81–86% of the maximum theoretical yield). Although the rate of ethanol production increased with cellulase, diminishing improvements were observed at enzyme loadings above 10 filter paper units/g of cellulose. Over 40 g of ethanol/L was produced with relatively low enzyme loadings: 7–10 filter paper units/g of cellulose. Two optimal SSF conditions were identified for fermentation yield with strain P2: pH 5.2 at 35 °C and pH 5.5 at 32 °C. Under these conditions, 47 g of ethanol/L was produced in 144 h (0.48 g of ethanol/g of cellulose). Maximal rates of ethanol production were observed at 37 °C and pH 5.0 and produced over 40 g of ethanol/L in 72 h (final yield of 0.432 g of ethanol/g of cellulose after 96 h). All fermentations except those conducted at 40 °C and low pH (pH 5.0–5.5) exceeded 70% of theoretical ethanol yields. Tight process controls may not be required using this organism since temperatures between 32 and 37 °C at pH ranges between 5.0 and 5.8 still produced good yields.

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“…SSF performance was improved by the yeast Brettanomyces custersii, which ferments cellobiose directly into ethanol, or by coculturing the less-ethanoltolerant yeast B. claussenii with the more robust Saccharomyces yeast (Spindler et al 1992). Similar benefits are provided by bacteria genetically engineered to ferment xylose (one of the sugars derived from hemicellulose) into ethanol, bacteria that ferment cellobiose into ethanol either naturally or through genetic modifications, and organisms that also make cellulase components (Wood and Ingram 1992).…”

Section: Saccharification and Fermentationmentioning

confidence: 80%

Cellulosic biomass could help meet California's transportation fuel needs

Wyman¹,

Yang²

2008

Cal Ag

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Evaluation of the cellobiose-fermenting yeastBrettanomyces custersii in the simultaneous saccharification and fermentation of cellulose

Cited by 59 publications

References 12 publications

Assessing Genetic Diversity among Brettanomyces Yeasts by DNA Fingerprinting and Whole-Genome Sequencing

Assessing Genetic Diversity among Brettanomyces Yeasts by DNA Fingerprinting and Whole-Genome Sequencing

Fermentation of Crystalline Cellulose to Ethanol by Klebsiella oxytoca Containing Chromosomally Integrated Zymomonas mobilis Genes

Cellulosic biomass could help meet California's transportation fuel needs

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