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Brettanomyces

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Unlike most genres of yeast, ''Brettanomyces'' has the characteristics of being very tolerant to harsh conditions, including high amounts of alcohol (up to 14.5-15% ABV <ref name="Crauwels1" /><ref name="Agnolucci_2017" />), a pH as low as 2 <ref>[http://www.winesandvines.com/template.cfm?section=news&content=141954 Wines and Vines. New Research on Role of Yeast in Winemaking; report on a presentation by David Mills and Lucy Joseph from UC Davis. 11/14/2014. Retrieved 08/16/2015.]</ref>, and environments with low nitrogen <ref name="Schifferdecker"></ref>. It has been reported that some strains require a very low concentration of fermentable sugars (less than 300 mg/L) and nitrogen (less than 6 mg/L), which is less than most wines contain <ref name="Smith_2017">[https://www.sciencedirect.com/science/article/pii/S0740002017308249 The carbon consumption pattern of the spoilage yeast Brettanomyces bruxellensis in synthetic wine-like medium. Brendan D. Smith, Benoit Divol. 2017.]</ref>. This capability allows ''Brettanomyces'' to survive in alcoholic beverages such as beer, wine, and cider. In alcoholic beverages, ''B. bruxellensis'' tends to lag after the primary fermentation with ''Saccharomyces''. It is believed that during this lag phase, ''B. bruxellensis'' adapts to the harsh conditions of the beverage (low pH, high concentrations of ethanol, and limited sugar/nitrogen sources). After this lag phase, ''B. bruxellensis'' can grow and survive when no other yeasts can. ''Brettanomyces'' is also more resistant to pH and temperature changes, and tolerant of environments limited in oxygen (although ''Brettanomyces'' prefers the availability of at least a little bit of oxygen). Scientifically, which specific nitrogen and carbon sources ''B. bruxellensis'' uses in these stressful environments has not received much research <ref name="smith_divol_2016"></ref>.
''Brettanomyces'' is known for not producing much glycerol in beer. [https://en.wikipedia.org/wiki/Glycerol Glycerol] is a colorless, sweet-tasting, viscous liquid that is an important contributor to the mouthfeel of beer. Glycerol is produced as a stress response by a wide range of microbes, including ''S. cerevisiae'', and various species and strains of ''Debaryomyces'', ''Candida'', ''Lachancea'', and ''Zygosaccharomyces''. Despite not producing amounts of glycerol that are perceivable in beer, some strains of ''Brettanomyces bruxellensis'' actually produce glycerol which is stored inside of their cells as a response to osmotic stress. They can also uptake glycerol into their cells. Doing so allows the cells to survive osmotic pressure <ref>[https://www.sciencedirect.com/science/article/pii/S0740002013001251?via%3Dihub Osmotic stress response in the wine yeast Dekkera bruxellensis. Silvia Galafassi, Marco Toscano, Ileana Vigentin, Jure Piškur, Concetta Compagno. 2013.]</ref><ref>[https://academic.oup.com/femsle/advance-article-abstract/doi/10.1093/femsle/fny020/4828327?redirectedFrom=fulltext Osmotolerance of Dekkera bruxellensis and the role of two Stl glycerol-proton symporters. Jana Zemančíková, Michala Dušková, Hana Elicharová, Klára Papoušková, Hana Sychrová. 2018</ref>. It is currently not known how many strains are capable of producing glycerol internally, or if this amount of glycerol has any impact on perceived mouthfeel of a beer if a substantial amount of ''Brettanomyces'' cells eentually eventually autolyze (see [https://www.facebook.com/groups/MilkTheFunk/permalink/2003626776332193/ this MTF thread]). The role of glycerol in creating mouthfeel is debatable in the wine world <ref>[https://www.winesandvines.com/features/article/68760 Tim Patterson. "Many Roads to Mouthfeel". Wines & Vines Magazine. Nov 2009. Retrieved 03/23/2018.]</ref>.
The genetic diversity of ''Brettanomyces'' is particularly wide. Some studies have indicated that strains of ''B. bruxellensis'' have adapted to specific environments. For example, one study found that strains of ''B. bruxellensis'' isolated from wine had 20 genes involved in the metabolism of carbon and nitrogen, whereas strains isolated from beer did not. This indicated that ''B. bruxellensis'' strains living in wine have adapted to the harsher environment of wine <ref name="smith_divol_2016"></ref>. Another study found that one out of the two strains tested that were isolated from soda could not ferment maltose, and only strains isolated from wine were able to grow in wine and the beer/soda strains did not. The wine strains were also more resistant to sulfites, which are commonly used in the wine industry to prevent microbial contamination <ref name="Crauwels_2016" />.
A genetic survey of 1488 different strains of ''B. bruxellensis'' from 29 countries, 5 continents, and 9 different fermentation niches was conducted in 2018 by Avramova et al. They found that these strains formed roughly 5 genetic groups with mostly separate ancestral lineages, and 1 group with a mixed ancestral lineage: 3 wine groups, 1 beer group, 1 kombucha group (most distantly related to the beer group), as well as 1 tequila/ethanol group that has multiple ancestral lineages. This was expressed mostly in the ploidy level of each group (the number of sets of chromosomes), with 2 of the wine groups, the tequila group, and the beer group containing more sets of chromosome pairs than the other groups (diploid vs triploid; this is thought to encourage adaption and hybridization). Additionally, the triploid wine group was generally more tolerant of SO<sup>2</sup> than the diploid wine groups. The genetic differences between the fermentation substrates (beer, wine, etc.) were lower but still significant, and this was explained by the frequent cross-over of equipment such as wine barrels being used for beer fermentation. When comparing the geographic differences, they found geography contributed only 5% of genetic differences, while geography explained more than 50% of genetic differences in non-wine strains, suggesting that beer, kombucha, and tequila strains are more localized genetically than wine strains, and that humans probably helped the wine strains travel across the globe. They also found that although one study reported spore-forming versions of ''B. bruxellensis'' (referred to as ''Dekkera bruxellensis''), the genetic makeup of the analyzed strains determined their ability to sporulate to be non-existent or rare (only one study that we know of by Walt and Kerken in 1960 has reported sporulation in ''Brettanomyces'', indicating that sporulation in ''Brettanomyces'' is extremely rare) <ref>[https://www.nature.com/articles/s41598-018-22580-7 Brettanomyces bruxellensis population survey reveals a diploid-triploid complex structured according to substrate of isolation and geographical distribution. Marta Avramova, Alice Cibrario, Emilien Peltier, Monika Coton, Emmanuel Coton, Joseph Schacherer, Giuseppe Spano, Vittorio Capozzi, Giuseppe Blaiotta, Franck Salin, Marguerite Dols-Lafargue, Paul Grbin, Chris Curtin, Warren Albertin & Isabelle Masneuf-Pomarede. 2018. doi:10.1038/s41598-018-22580-7.]</ref>. See also [https://www.facebook.com/groups/MilkTheFunk/permalink/2022801681081369/ Richard Preiss's discussion of this study on MTF].
Sulfite and SO<sub>2</sub> inhibits the growth of ''Brettanomyces'', and is often used in the wine industry to prevent the growth of ''Brettanomyces'' (some wineries have identified small amounts of flavors from ''Brettanomyces'' as being beneficial to certain wine styles, and is said to increase the complexity and impart an aged character in young wines <ref name="smith_divol_2016"></ref>) <ref>[http://onlinelibrary.wiley.com/doi/10.1111/j.1745-4549.2012.00702.x/abstract Removal of Brettanomyces Bruxellensis from Red Wine Using Membrane Filtration. Umiker, Descenzo, Lee, and Edwards. 04/24/2012.]</ref>. However, it has been shown that wine strains of ''B. bruxellensis'' could survive dosages of up to 1 mg/L of SO<sub>2</sub>, and the very high dosage of 2.1 mg/L was needed to kill ''Brettanomyces'' in wine <ref name="Agnolucci_2017" />. This dosage of SO<sub>2</sub> is beyond legal limits and has negative effects on wine. The tolerance of SO<sub>2</sub> of strains of ''Brettanomyces'' found in beer is lower than those found in wine strains. Some strains of ''Candida pyralidae'', ''Wickerhamomyces anomalus'', ''Kluyveromyces wickeramii'', ''Torulaspora delbrueckii'' and ''Pichia membranifaciens'' have been found to produce toxin that inhibits ''Brettanomyces'', and these toxins have been proposed as an alternative to SO<sub>2</sub> as a way to kill ''Brettanomyces'' (killer wine strains of ''Saccharomyces cerevisiae'' do not kill ''Brettanomyces''; see [[Saccharomyces#Killer_Wine_Yeast|Killer Wine Yeast]] for more information).

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