Nonconventional Yeasts and Bacteria
Nonconventional Yeasts and Bacteria are yeasts and bacteria genre that haven't been greatly explored in alcoholic fermentation, but might prove to be worth exploration. This page contains anecdotal information, as well as scientific information that might prove useful for brewers who are looking to brew with microbes that don't include the typical lab yeasts and bacteria for sour/mixed fermentations. For yeasts and bacteria that are more often used in sour and mixed fermentations, see Saccharomyces, Brettanomyces, Lactobacillus, and 'Pediodoccus.
Under progress
Potential references:
- https://www.facebook.com/groups/MilkTheFunk/permalink/1336235339738010/?comment_id=1336277939733750&comment_tracking=%7B%22tn%22%3A%22R%22%7D - https://www.facebook.com/groups/MilkTheFunk/permalink/1337089182985959/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1346900285338182/ - http://www.sciencedirect.com/science/article/pii/S0963996916302332 - https://www.facebook.com/groups/MilkTheFunk/permalink/1366829093345301/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1365795896781954/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1380004022027808/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1284664904895054/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1400174630010747/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1420821137946096/ - http://www.sciencedirect.com/science/article/pii/S074000201630452X - https://www.facebook.com/groups/MilkTheFunk/permalink/1457271340967742/ - Review: Pure non-Saccharomyces starter cultures for beer fermentation with a focus on secondary metabolites and practical applications - https://www.facebook.com/groups/MilkTheFunk/permalink/1485339661494243/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1140282595999953/ - https://www.ncbi.nlm.nih.gov/pubmed/12102552 - https://www.facebook.com/groups/MilkTheFunk/permalink/1546044102090465/ - http://beer.suregork.com/?p=3860 - http://ijs.microbiologyresearch.org/content/journal/ijsem/10.1099/ijsem.0.001607 - https://www.facebook.com/groups/MilkTheFunk/permalink/1582089058485969/ - http://www.mbaa.com/publications/tq/tqPastIssues/2017/Pages/TQ-54-1-0215-01.aspx - http://biorxiv.org/content/early/2017/03/27/121103 - https://www.facebook.com/groups/MilkTheFunk/permalink/1640324282662446/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1659004047461136/ - https://www.facebook.com/groups/MilkTheFunk/permalink/1669790909715783/ - http://onlinelibrary.wiley.com/doi/10.1002/jib.381/full - http://www.asbcnet.org/publications/journal/vol/2017/Pages/ASBCJ-2017-2532-01.aspx - https://www.facebook.com/groups/MilkTheFunk/permalink/1680093658685508/ - http://onlinelibrary.wiley.com/doi/10.1002/yea.3146/abstract - https://mail.google.com/mail/u/1/?ui=2&ik=1b8e47c65b&view=att&th=15c548b66cda8ae6&attid=0.1&disp=safe&zw - https://www.facebook.com/groups/MilkTheFunk/permalink/1649825158379025/
Yeasts
Toluraspora Delbrueckii
Toluraspora Delbrueckii is species of yeast, that is round to ovoid in shape and has been traditionally used in some wine fermentations to increase the complexity. Most of the commercial Torulaspora species and strains were isolated from soil, fermenting grapes (wine), berries, agave juice, tea-beer, apple juice, leaf of mangrove a tree, moss, lemonade and tree barks. Although it was said that most T Delbrueckii strains would not fully attenuate or tolerate higher alcohol contents it has been shown that is strain dependent.
General Information
Analysis was done on 10 different T Delbruckii strains on various types of resistances as well as the ability to metabolize different carbon sources. The strains tested and the results are shown below.
| Designation | Strain number/signature | Origin |
|---|---|---|
| T6 | RIBMa TdA | Wine |
| T9 | DSMb 70504 | Sorghum Brandy |
| T10 | CBSc 1146T | Unknown |
| T11 | TUMd 214 | Bottle (Pils beer, trace contamination, no beer spoilage observed) |
| T13 | TUMd TD1 | Wheat beer (starter culture) |
| T15 | TUMd 138 | Cheese brine |
| T17 | WYSC/Ge 1350 | Unknown |
| T18 | CBSc 4510 | Unknown |
| T19 | DSMb 70607 | Unknown |
| T20 | CBSc 817 | Unknown |
Hop Resistance
The resistance to alpha acids were also measured among these 10 strains using 0 PPM, 50 PPM, and 90 PPM. All strains were found to be resistant to these levels of alpha acids not affecting their growth. Some strains however were shown to have slower growth rates in the presence of 90 PPM and more.
Ethanol Resistance
All 10 strains were also tested for their ability to grow in 5-10% ethanol content. The table below shows that all but one strain was able to grow in presence of 5% total alcohol but one thing they all shared in common is their inability to grow when in the presence of 10% alcohol.
Growth (+) positive; (−) negative.
| Ethanol % | T6 | T9 | T10 | T11 | T13 | T15 | T17 | T18 | T19 | T20 |
|---|---|---|---|---|---|---|---|---|---|---|
| 5% | - | + | + | + | + | + | + | + | + | + |
| 10% | - | - | - | - | - | - | - | - | - | - |
Cross Resistance
Again, all 10 strains growth was tested but this time with the presence of both 5% ethanol as well as 50 and 90 PPM of iso-alpha acid concentrations. Below you can see that with a combination of these two factors, growth was hindered in quite a few strains.
Growth (+) positive; (−) negative.
| IBU/ethanol % (v/v) | T6 | T9 | T10 | T11 | T13 | T15 | T17 | T18 | T19 | T20 |
|---|---|---|---|---|---|---|---|---|---|---|
| 50/5 | - | + | - | + | - | + | + | + | + | - |
| 90/5 | - | - | - | + | - | + | + | + | + | - |
Pichia
Bacteria
Oenococcus
Oenococcus Kitaharae
O. kitaharae is a lactic acid bacterium (LAB) that was isolated from composting distilled shochu residue produced in Japan. This species represents only the second member of the genus Oenococcus to be identified. O. kitaharae has the ability to ferment maltose, citrate and malate and the ability to synthesize specific amino acids such as L-arginine and L-histidine unlike some O. Oeni. In addition to these metabolic differences, the O. kitaharae genome also encodes many proteins involved in defense against both bacteriophage (restriction-modification and CRISPR) and other microorganisms (bacteriocins), and has had its genome populated by at least two conjugative transposons, which is in contrast to currently available genome sequences of O. oeni which lack the vast majority of these defense proteins. It therefore appears that the genome of O. kitaharae has been shaped by its need to survive in a competitive growth environment that is vastly different from that encountered by O. oeni, where environmental stresses provide the greatest challenge to growth and reproduction. [1]
Sugar Utilization -
One of the defining biochemical differences between O. kitaharae and O. oeni that was noted in its original isolation was the ability of O. kitaharae to produce acid from maltose. This trait is rare in O. oeni, which is formally classified as maltose negative. By comparing available whole-genome annotations for O. oeni with O. kitaharae, it was possible to identify several genes associated with sugar utilization that are deferentially present across the species. Of these, at least four genes which are present in O. kitaharae, but absent in the O. oeni genomes, are predicted to be involved in the utilization of maltose, providing a direct genetic basis for this phenotype. In addition to genes predicted to be involved in the species-specific utilization of maltose, there are several ORFs predicted to be involved in the metabolism of trehalose, D-gluconate, D-ribose and fructose that are specifically present in O. kitaharae. While the assimilation of these sugars is often carried out by specific strains of O. oeni, this genotypic data agrees well with biochemical tests performed previously that indicated that O. kitaharae was able to utilize all of these various carbon sources. [2]
Oenococcus oeni
Oenococcus oeni(also know as Leuconostoc oeni) is a Genus of Gram-positive LAB, ellipsoidal to spherical in shape that is primarily used in Malolactic Fermentation. Oenococcus oeni is a facultative anaerobe. It is able to use oxygen for cellular respiration but can also gain energy through fermentation. It characteristically grows well in the environments of wine, being able to survive in acidic conditions below pH 3.0 and tolerant of ethanol levels above 10%. Optimal growth occurs on sugar and protein rich media. Cells tend to grow in chains or pairs. O. Oeni is heterofermentative and generally produces CO2, Ethanol, Acetate, and Diacetyl.
O. oeni ferments sugars using both the hexose-monophosphate and phosphoketolase pathways using the enzymes Glucose-6-phosphate and xylulose-5-phosphoketolase to from D(-)-lactic acid, CO2 and ethanol in equal amounts when metabolising D-glucose. O.oeni can convert pentose phosphate to acetic acid in an oxygen dependant reaction which requires NADP. It cannot metabolize polysaccharides and alcohols.
O. oeni can decarboxylate L-malate to L(+)-lactate, but cannot use it as a sole source of carbon. It requires the amino acids Glutamic acid, valine, guanine, adenine, xanthine, uracil, riboflavin, folic acid, nicotinic acid, thiamine, biotine and pantothenic acid. There is some variation of amino acid requirement between strains.
Althought O. Oeni has primarily been used for Malolactic Fermentation, trials with the White Labs culture(only one reported on so far) has show lactic acid production without the presence of malic acid. [3] James Sites reported souring within a week at 70F.
| Name | Mfg# |
|---|---|
| White Labs | Malolactic Culture |
| Wyeast | Malolactic Blend |
| CHR Hansen | Viniflora |
Weisella
See also: