High ester yeast for whiskey

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Re: High ester yeast for whiskey

Postby cuginosgrizzo » Thu Nov 02, 2017 6:27 am

Lyonsie wrote:Im actually intrested to have come across this thread. I have 200l of apple juice getting pressed this Saturday. And although i dont really understand the science behind it i have purchased two yeasts that are noted for the production of high esters and fusil oils, in a bid to carry over apple flavour as most of this is in the heads.
The two varieties i chose are ma33 and aw4. Both belong too the Saccharomyces Cerevisiae family. Im expecting a starting abv of about 6% and it will be ran slowly through a pot and a thumper loaded with a mixture of wash and concentrated apple syrup. If anyone with more experience thinks im going wrong here id be grateful for any input.


Hi Lyonsie, I am no expert at all but what I learnt on this forum is that Saccharomyces Bayanus works better with apple juice, as they are cleaner. You don't want to cover the apple flavour with "alien" esters from the yeast.
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Re: High ester yeast for whiskey

Postby OtisT » Mon Nov 06, 2017 10:57 am

RedwoodHillBilly wrote:
kiwi Bruce wrote:I read recently, (Tummy bug that laid me up for a week, get lots of reading done) that the esters yeasts produce are only partially released into the wort, the bulk are stored internally...and released by boiling.


Interesting, have you got a reference? If this is so, my method of steam stripping the grains and trub using clear beer in the boiler has yet another justification. I do believe that I get more grain flavor this way, but I hadn't thought that I'd get more esters as well.


+1 to Interesting and finding the source.
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Re: High ester yeast for whiskey

Postby kiwi Bruce » Thu Nov 09, 2017 5:53 am

I'll find the article again eventually, I'll need to do some mining though...my son rebuilt my computer while I was laid up and my "history" is in the cloud somewhere...and not in a cloud of still vapor, I could find that!
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Re: High ester yeast for whiskey

Postby Jimy Dee » Wed Jan 03, 2018 8:20 pm

Gents, what a great thread. I look forward to all your conclusions in the fullness of time. I am realising again that this hoby contains deep running water and will take years of study to move out of novice category. Jimy
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Re: High ester yeast for whiskey

Postby zapata » Thu Jan 04, 2018 3:29 am

Found this
The ester content of distilled beverages is greatly dependent on whether or not the yeast is present at the time of distillation. We have found that the distillation
in the presence of yeast increases the amount of the ethyl esters of caprylic, capric and palmitoleic acids.33

J. Insl. Brew., May-June, Vol. 85, pp. 149-156 149
THE PRODUCTION OF AROMA COMPOUNDS BY YEAST
By Heikki Suomalainen* and Matti Lehtonen
Found pdf easily on Google, but copying a link didn't want to work.
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Re: High ester yeast for whiskey

Postby zapata » Thu Jan 04, 2018 3:36 am

Actually, I chased the footnote and it directly answers the question, so here it is.
DISTRIBUTION OF ESTERS PRODUCED DURING SUGAR FERMENTATION BETWEEN
THE YEAST CELL AND THE MEDIUM
By Lalli Nykanen, Irma Nykanen and Heikki Suomalainen
The proportion of the fatty acid ethyl esters transferred to the medium decreased
with increasing chain length: all in the medium for ethyl caproate, 54-68% for ethyl caprylate, 8-17%
for ethyl caprate, and all remaining in the yeast cell for ethyl laurate.

j.2050-0416.1975.tb03788.x.pdf
(260.44 KiB) Downloaded 26 times
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Re: High ester yeast for whiskey

Postby zapata » Thu Jan 04, 2018 3:59 am

And one more, but can anybody get their hands on the full text of this article? They apparently do tests with safale s04 under a variety of variables, so it should directly relate to the interests of this thread.

Hiralal, L., Olaniran, A. O., & Pillay, B. (2014). Aroma-active ester profile of ale beer produced under different fermentation and nutritional conditions. Journal of Bioscience and Bioengineering, 117(1), 57-64. doi:10.1016/j.jbiosc.2013.06.002

This ph idea was recently posted in a rum thread from the lost spirits distiller, but here it is again from a beer researcher summarizing Hiralal:
Beer pH
The Hiralal study also looked at the role of beer pH and its effect on ester production, which is something I hadn’t seen studied before. Three different beers were fermented each with a different starting pH (3.0, 5.0, and 7.0). As the starting pH increased, so did the esters formed during fermentation, but a low starting pH actually reduced the esters. The beer fermented at a pH of 7.0 resulted in the highest ester concentration with a 13% increase in total acetate esters and 7% increase in total ethyl ester production compared to the control (5.0 pH beer). On the other hand, the beer fermented at a low 3.0 pH resulted in an 18% decrease in total ester concentration.

So maybe bumping that ph up isn't a bad idea, even for a known estery strain?
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Re: High ester yeast for whiskey

Postby Single Malt Yinzer » Thu Jan 04, 2018 4:20 am

Good find Zapata. The pH thing is interesting. I have no idea why, making it a good area to research.

Some of the other work we've quoted was from Heikki Suomalainen also. I'll have to check into him and see what else I can find.

Found this: https://www.amazon.com/Distilled-Alcoho ... 902771553X

Anyone want to buy me a late Xmas gift? :twisted:
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Re: High ester yeast for whiskey

Postby OtisT » Thu Jan 04, 2018 10:26 am

Single Malt Yinzer wrote:Good find Zapata. The pH thing is interesting. I have no idea why, making it a good area to research. :


A few other active threads are discussing the PH effect on esterfication, and some science on why this works. One thread I recall is Redwood Hillbilly’s “using pure acid for esterfication” thread. Also several others where Der Wo and others discuss acids used for esterfication. Use HD Google search for pure acid, or sulfuric acid, or similar to fine some of these.

I just bought a liter of 98% pure sulfuric acid that I will be experimenting with on my next runs. ( thanks RHB for the link to the acid seller online). Added to low wines ( I will also mix with backseat and feints) and a longer wash warmup procedure, it is said to dramatically impact esterfication.

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Re: High ester yeast for whiskey

Postby Single Malt Yinzer » Thu Jan 04, 2018 10:45 am

Not the acids thing, the observation that starting with a mash pH of 7.0 increased ester production over 5.0 and 3.0. It's the opposite of what I think we would have thought. Most of us aim for a pH of 5-5.5 for mashing for max enzyme activity. I don't think anyone then corrects pH to 7, they leave it as it was at the end of the mash.
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Re: High ester yeast for whiskey

Postby RedwoodHillBilly » Thu Jan 04, 2018 10:49 am

OtisT wrote:I just bought a liter of 98% pure sulfuric acid that I will be experimenting with on my next runs. ( thanks RHB for the link to the acid seller online). Added to low wines ( I will also mix with backseat and feints) and a longer wash warmup procedure, it is said to dramatically impact esterfication.
Otis


Otis, please let us know the results of your experiments. Backset along with H2SO4 in your low wines should give should you some interesting esters. Now, whether they are what you are looking for is a different story.

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Re: High ester yeast for whiskey

Postby RedwoodHillBilly » Thu Jan 04, 2018 10:52 am

Single Malt Yinzer wrote:Not the acids thing, the observation that starting with a mash pH of 7.0 increased ester production over 5.0 and 3.0. It's the opposite of what I think we would have thought. Most of us aim for a pH of 5-5.5 for mashing for max enzyme activity. I don't think anyone then corrects pH to 7, they leave it as it was at the end of the mash.


I wonder what the mechanism for that is? Bacterial infection or yeast metabolism or ?????
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Re: High ester yeast for whiskey

Postby zapata » Thu Jan 04, 2018 12:31 pm

It is yeast metabolism in this case (though bacteria play their own separate role). The main chemical ester pathway is thought to be the Fischer-Speier reaction. Carboxylic acid + alcohol -> esters (sped up by heat and catalysts like sulphuric acid). This is the route that no doubt happens in the barrel (slowly), in the boiler, dunder pit (slowly), and certainly with pure H2SO4.

The yeast metabolic ester pathway is also very well documented. It is an internal yeast process governed by yeast enzymes (acyl transferases), and at least for the acetate esters "we" know the specific enzymes and even the exact genes that produce and regulate them. There may be individual genes and enzymes for each ester (or at least types of esters) which would be the genetic reason some yeast strains are more estery than others. These esters are the esters stemming from yeast selection and fermentation conditions like ph, temperature, pressure, oxygen, yeast pitch rate etc.
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Re: High ester yeast for whiskey

Postby Single Malt Yinzer » Thu Jan 04, 2018 12:56 pm

And another rabbit to go down: https://academic.oup.com/femsyr
FEMS Yeast Research is the leading yeast community journal, bridging the gap between pure and applied research on yeasts.


https://academic.oup.com/femsyr/article ... 005/534737 - Effects of high medium pH on growth, metabolism and transport in Saccharomyces cerevisiae

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1275219/ - pH values of the yeast cell

I didn't read these yet, but they should help maybe understand how pitching pH affects yeast and ester production.
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Re: High ester yeast for whiskey

Postby zapata » Thu Jan 04, 2018 3:13 pm

I looked through that 1st one "Effects of high medium pH on growth, metabolism and transport in Saccharomyces cerevisiae "
It seems mainly focused on the reduced growth (not life or metabolism in general) of yeast at ph well above 7, but this is what I gleaned that seems relevant.
1. Yeast actively acidify wort by actively pumping H+ ions out of the cell. This acidification is almost nonexistant at ph 4, but increases significantly as the ph rises, even at ph 6 though they measured it up to ph 9.
2. The increased acid production at higher ph was mostly due to increased co2 production and thus carbonic acid. But a significant amount of an unidentified organic acid was also produced. Citing prior research it was proposed this may be succinic acid. (My comment: Succinic acid is a carboxylic acid and can thus end up as an ester. Esters of succinic acid are foynd naturally in alcoholic beverages particularly aged ones. An example is diethyl succinate which is described as fruity tart tropical floral passion fruit, though I only very briefly looked into succinic esters in general)
3. Uptake of amino acids (specifically C-leucine) by yeast was increased as ph increased from 4 to 6 to 8. It decreased at ph 9. (My extrapolation, but given my newfound knowledge of yeast metabolism of amino acids into fusel alcohols, this could well cause an increase in non-ethyl esters, and/or fusel s themselves if not esterized).
4. There was not a difference in the slowed growth at ph 9 when using either NaOH or KOH to maintain high ph. (My comment, but a general stillers knowledge thing I have heard is that K is better for yeast than Na, so many prefer to adjust ph up with K bases instead of Na bases. At least from this study, and in terms of growth alone that practice does not seem validated).
5. I got a bit lost at this point, but it seems overall yeast health and fermentation abilities were better for yeast previously incubated at ph 4. (My deduction is yeast starters should be ph 4, even if the wort is higher)
6. Quick swings to higher ph showed increased pyruvate kinase reactions. This is the enzyme responsible for the final step of converting glucose to pyruvate and producing ATP for cellular energy in the process.
(My comment; both ATP and pyruvate are integral steps in separate pathways for the production of esters by yeasts. See the following quote from an earlier article:
Esters are formed via an intracellular process, catalysed by an acyl transferase or ‘ester synthase’ (Nordström, 1962). The reaction requires energy provided by the thioester linkage of the acyl‐CoA co‐substrate (Fig. 2). The most abundant acyl‐CoA is acetyl‐CoA, which can be formed either by oxidative decarboxylation of pyruvate or by direct activation of acetate with ATP. The majority of acetyl‐CoA is formed by the oxidative decarboxylation of pyruvate, while most of the other acyl‐CoAs are generated by the acylation of free CoA catalysed by acyl‐CoA synthase (fatty acid metabolism).


TL/DR
Overall takeaway seems to be that ph well above 4 definitely affects known processes leading to ester formation from both chemical and metabolic pathways, and if the ph is only 6 or 7 this can be maximised without many negative affects, especially if yeast is previously grown at ph 4 as in a traditionally acidic starter.
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Re: High ester yeast for whiskey

Postby OtisT » Thu Jan 04, 2018 6:13 pm

Single Malt Yinzer wrote:Not the acids thing, the observation that starting with a mash pH of 7.0 increased ester production over 5.0 and 3.0. It's the opposite of what I think we would have thought. Most of us aim for a pH of 5-5.5 for mashing for max enzyme activity. I don't think anyone then corrects pH to 7, they leave it as it was at the end of the mash.


Oops. My bad misreading that. Sorry to bring this off topic. Otis
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Re: High ester yeast for whiskey

Postby OtisT » Thu Jan 04, 2018 6:30 pm

RedwoodHillBilly wrote:Otis, please let us know the results of your experiments. Backset along with H2SO4 in your low wines should give should you some interesting esters. Now, whether they are what you are looking for is a different story.

RHB


You bet I will, in the pure acid thread. It will be a while though, maybe two more months, and my observations will be even more subjective than they normally are. I’m in the process of changing a lot of equipment out right now :D so while I plan to start with a recipe I know, HBB, the new equipments will have some impact on the results. I also have infected backset and dunder that’s been stewing for months, so that should be quite interesting In combination with the acid. I’m so excited, I can’t wait. :D

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Re: High ester yeast for whiskey

Postby Single Malt Yinzer » Fri Jan 05, 2018 4:43 am

OtisT wrote:Oops. My bad misreading that. Sorry to bring this off topic. Otis


No worries, it's not like this topic is overly complicated or anything. :)
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Re: High ester yeast for whiskey

Postby Shine0n » Fri Jan 05, 2018 5:58 pm

Single Malt Yinzer wrote:
OtisT wrote:Oops. My bad misreading that. Sorry to bring this off topic. Otis


No worries, it's not like this topic is overly complicated or anything. :)

I don't know, all this has made my brain hurt!
You cats are well above my understanding of things, lol

I've done different temps with different yeasts to find their best points of esters and what they would add to spirits and obviously the dunder threads but you guys are bringing things to a higher level.

Keep up the good work!
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Re: High ester yeast for whiskey

Postby Single Malt Yinzer » Thu Jan 11, 2018 9:58 am

Parameters Affecting Ethyl Ester Production by Saccharomyces cerevisiae during Fermentation - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2223249/

We concluded that, at least in our fermentation conditions and with our yeast strain, the fatty acid precursor level rather than the activity of the biosynthetic enzymes is the major limiting factor for ethyl ester production. The expression level and activity of the fatty acid biosynthetic enzymes therefore appear to be prime targets for flavor modification by alteration of process parameters or through strain selection.


Fatty acids and their precursors are found in backset (Dunder & Sour Mashing) as well as tails. So another factor in yeast selection is fatty acid production? I'm don't believe that is openly measured by the industry as I don't believe it's something that brewers/distillers look for in a yeast.

Yeast selection criteria so far I see for esters (no other factors) is:
1. aCoA or ATF/AAT (Ester producing enzymes)
2. Fatty acid production
3. Higher alcohol production (fusel alcohols)

Please in in your thoughts. I could and mostly likely be off the deep end. :)
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Re: High ester yeast for whiskey

Postby zapata » Thu Jan 11, 2018 2:36 pm

That is an interesting find for several reasons. For one they actually used an AG malt barley wort. Also, some homebrewers have been advocating the use of small amounts of oil in fermentation for a while. Usually olive oil and usually in lieu of (or addition to) oxygenation though. For what it is worth, the paper specifies the fatty acids they supplemented with, and a quick glance shows soybean oil is the closest in FA makeup to that used in the study, though it wasn't clear to me if the choice of fatty acids can tailor the esters produced.

I had previously thought that selecting an oil used for anti-foam could affect chemically (fischer-spier) derived esters in the boiler. TBD on that, but looks like for sure an oil supplement before fermentation will have an effect via yeast metabolised esters. Cool.
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Re: High ester yeast for whiskey

Postby Single Malt Yinzer » Thu Jan 11, 2018 6:21 pm

Nice list of them here:
https://en.wikipedia.org/wiki/Saturated_fat

Coconut oil and Palm kernel oil top the list by a lot. Butter is pretty high tool. Hmmm... Buttered Bourbon? :)
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Re: High ester yeast for whiskey

Postby butterpants » Fri Jan 26, 2018 12:38 am

RedwoodHillBilly wrote:I started this thread as a place to discuss using high ester producing yeasts in whiskey ferments. We were starting to get off topic in the pure acids for esters thread. https://homedistiller.org/forum/viewtop ... 0#p7485884

Let the discussion begin.
zapata wrote:That is an interesting find for several reasons. For one they actually used an AG malt barley wort. Also, some homebrewers have been advocating the use of small amounts of oil in fermentation for a while. Usually olive oil and usually in lieu of (or addition to) oxygenation though. For what it is worth, the paper specifies the fatty acids they supplemented with, and a quick glance shows soybean oil is the closest in FA makeup to that used in the study, though it wasn't clear to me if the choice of fatty acids can tailor the esters produced.

I had previously thought that selecting an oil used for anti-foam could affect chemically (fischer-spier) derived esters in the boiler. TBD on that, but looks like for sure an oil supplement before fermentation will have an effect via yeast metabolised esters. Cool.
That oil over O2 was debunked a while back. I've never seen anyone use it commercially.... if I remember correctly it was derived by a paper written at New Belgium brewing in CO. It was just pillow talk. They do not do this now.
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Re: High ester yeast for whiskey

Postby VLAGAVULVIN » Sat Feb 10, 2018 9:31 am

kiwi Bruce wrote:One of the intriguing things I found in whisky science magazine, was the length of fermentation time that the heavily estered/phenolic single malt take. Oban, Ardbeg, laphroaig and lagavulin can let their ferments run ninety hours (I believe the average Spayside is around thirty hours)...s.

Can't believe it. In Russia we say "u durakov mysli odinakovye" (fools share their mind on air). I have the same article been opened maybe since Feb.'2017 (and it's opened just now). On one hand, the most of yeast is inactive after first 48 hours (being dead or sick, hehe). Due to its "sickness" (lo-sugars + hi-alco + progressing autolysis), the colony starts to make more and more by-products. But the U.S. tradition tends to have hi-nitro wort in the very beginning (sour mash or dunder or "rum bomb" + the non-filtered mash fermentation). And... I didn't taste much of American malt or corn based beverages but all I know of have put a long-long tail into my nose. In contrast to it, Islay is rich in "powerful" esters, maybe some feinty notes but not so definite tails. Longer aging only? Secret Gaelic strains? AFAIK, they filter the mash prior to pitching the yeast. No nitro-vitamin-bombs or porridge or backset inside. And SG 1.05..1.06. They make sorta "ale without hops", in fact. But still there is a suspected one.

kiwi Bruce wrote:To me that says they either have a bacterial ferment running and/or, the yeast strain needs extra time to make the profile their looking for.

Don't you think the suspect is lactobacteria? That "eats" some remaining sweet yummies (like dextrins, pentoses) and cell membranes of R.I.P.-ed yeast? Never used in your fermenter any starter culture for yogurt that we can buy just in foodstores? The colony would "piss" a lot of lactic acid (lowering pH to 3.8 by the day number four)... The acids bind fusels to esters, rite?

And besides... I found that my "peated/smoked" (phenolic) feints that I collect are not so stinky as "highland-like" mashed. In 3 months they positively changed just being put in a bottle. So, I took 50ml of them in a jar and dropped some oak chips. I'm getting the effect I am familiar with (as per my experiments with spent lees after my reflux column). I wonder if it's worth discussing it here or somewhere else...

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Re: High ester yeast for whiskey

Postby OtisT » Fri Mar 09, 2018 2:21 pm

I was very happy with the results when using this yeast in my recent batch of Honey Bear Bourbon. I fermented at 85 F and it finished dry in 5 days. I am happy with the taste of the cut I just put into a barrel. Lots of other factors went into the batch, so at a minimum the yeast did not hurt things.
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Re: High ester yeast for whiskey

Postby VLAGAVULVIN » Fri Mar 09, 2018 9:16 pm

OtisT wrote:I was very happy with the results when using this yeast


What was the total alcho % in your "beer" after it had finished to dry?

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Re: High ester yeast for whiskey

Postby OtisT » Fri Mar 09, 2018 9:55 pm

VLAGAVULVIN wrote:
OtisT wrote:I was very happy with the results when using this yeast


What was the total alcho % in your "beer" after it had finished to dry?


10 64 to 10 01 for 8.25%
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Re: High ester yeast for whiskey

Postby VLAGAVULVIN » Fri Mar 09, 2018 10:56 pm

Not bad yeast, thanks for your feedback.

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