Isolated Yeast (Tree House): How to Identify and Characterize?

[Lakeside_Brewer5]

Have you tested this yet, or are planning to?

I can’t speak for @troxerX but I’ll be testing the slightly simplified version I proposed in my original post. Should be starting it in a few weeks. I don’t have the professional setup @echoALEia has but I’ll focused on minimizing oxidation during the blending process.

Based on the use of Brite Tanks at TH, and how we’ve seen the randomness of pitching a yeast blend all at once, I’m not firmly in the camp multiple worts are being fermented and used until we have evidence otherwise.

 

[Noob_Brewer]

This time around I underpitched the SO4 and it’s smelling amazing so far. I’ll be dry hopping it on Sunday.

@echoALEia Ive asked this before somewhere in HBT with no response though. How do you know how many cells are in a packet of SO4 to know you are under pitching? Fermentis states is 6B cells per gram, but if you go by Mr. Malty standards, its 20B cells. So this is tremendous variation. What do you go by to know you are under pitching and what do you think your pitch rate actually is? Thanks.

 

[beervoid]

in-progress as I type but as I said, will not share details until I see others rolling up their sleeves.... @isomerization, @Clyde McCoy busted their ..... giving us the microbiology, now it’s time for others to get out of their ‘one wort’ world and help Joe @echoALEia and myself prove this right or wrong. No teamwork - no recipe...

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Fair enough, I'll def be joining. Been experimenting with copitching but havent had any cutting edge results.
I think we also need to focus more on how to retain esters in our worts. They are super volatile. I recently read and article that belgium brewers take the beer off the yeast as soon as possible in order to retain yeast esters as old yeasts absorb them. I wonder how this ties in with conditioning.
Especially considering these beers have tons of hops and enzymes that cause hop creep it seems tricky to get the beer off the yeast early as to prevent diacetyl from forming. Conditioning the beer with another yeast seems to me the answer to take care of the hop creep problem.
Unless they cheat and use ALDC

"One wort order" its a good one, made me laugh.

 

[TBryerton]

in-progress as I type but as I said, will not share details until I see others rolling up their sleeves.... @isomerization, @Clyde McCoy busted their ..... giving us the microbiology, now it’s time for others to get out of their ‘one wort’ world and help Joe @echoALEia and myself prove this right or wrong. No teamwork - no recipe...

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LOL. If you don’t want to share your recipe bud, don’t share your recipe. There’s been a lot of people who contributed to this thread, who shared beer, who shipped beer to be test, etc. You through out an idea that was also thrown out about 80 pages ago. Nothing wrong with that, but let’s not act like you’re the only one contributing.

 

[troxerX]

LOL. If you don’t want to share your recipe bud, don’t share your recipe. There’s been a lot of people who contributed to this thread, who shared beer, who shipped beer to be test, etc. You through out an idea that was also thrown out about 80 pages ago. Nothing wrong with that, but let’s not act like you’re the only one contributing.

As you can imagine by now I have pretty a good idea how whoever threw that idea 80 pages ago was ignored... we would probably have a good recipe by now... I’m about team work and cooperation and as you can see I have to get creative to convince the right people so we get results...

 

[TBryerton]

As you can imagine by now I have pretty a good idea how whoever threw that idea 80 pages ago was ignored... we would probably have a good recipe by now... I’m about team work and cooperation and as you can see I have to get creative to convince the right people so we get results...

As previously mentioned, I didn’t ignore. I was convinced that was the way to go and tried a couple batches. It made a good beer but wasn’t anything like TH. Very strong esters from the other 2 yeasts. They faded and became subtle very quickly. That being said, it was in my beginning stages of brewing, I didn’t have great oxygen control, was brewing in carboys. All things considered it made a good beer, but I wasn’t convinced this technique would produce anything with a TH profile.

 

[beervoid]

As previously mentioned, I didn’t ignore. I was convinced that was the way to go and tried a couple batches. It made a good beer but wasn’t anything like TH. Very strong esters from the other 2 yeasts. They faded and became subtle very quickly. That being said, it was in my beginning stages of brewing, I didn’t have great oxygen control, was brewing in carboys. All things considered it made a good beer, but I wasn’t convinced this technique would produce anything with a TH profile.

From being strong to fading out they never where even a bit close to TH esters?

 

[troxerX]

Fair enough, I'll def be joining. Been experimenting with copitching but havent had any cutting edge results.
I think we also need to focus more on how to retain esters in our worts. They are super volatile. I recently read and article that belgium brewers take the beer off the yeast as soon as possible in order to retain yeast esters as old yeasts absorb them. I wonder how this ties in with conditioning.
Especially considering these beers have tons of hops and enzymes that cause hop creep it seems tricky to get the beer off the yeast early as to prevent diacetyl from forming. Conditioning the beer with another yeast seems to me the answer to take care of the hop creep problem.
Unless they cheat and use ALDC

"One wort order" its a good one, made me laugh.

LOL ‘First Wort Order‘ —

Agreed, we need to understand more about esters, how they are formed and how to retain as much as possible.

With the blending approach, esters are taken care of, which is another advantage of the blending process. Larger Batch 1 ferments as usual, this batch purpose is to add the bread notes and base malt and serves as the primary hop platform, this batch will take the longest time as it needs to be dry hopped several times. Batch 2 is the ester batch with the ethyl-acetate (bubblegum) and isoamyl-acetate (banana). Per ‘BYO Hefeweizen’, fermentation for optimal notes should be done at 62F (slow ferm to avoid losing too many aromatics?) and with a higher ratio of glucose to maltose (here we go with the TH doesn’t use dextrose conversation again...). Maybe the simple sugars (glucose, dextrose, sucrose) aid in the formation of some esters? - anyway BYO doesn’t explain why. Per other references I’ve seen around, ethyl-acetate (bubblegum) is best produced with worts of higher gravities and with a grist of at least 65% wheat. Batch 2 once fermented it is quickly crashed, added CBC-1 for some time to kill T-58/WB-06 (to @Lakeside_Brewer5 point) and then quickly and carefully blended with the larger batch. By the time both batches are blended, batch 2 is the youngest one (if you are not pursuing batch 3) so my assumption is that the esters should still be there in sufficient concentration. Also if the gravity of the blend is high (i.e 16-18 points I would think the density may help retain esters?, anyway the important part is that the ester batch is fermented and blended after the larger batch went through the rigorous fermentation process.

 

[TBryerton]

From being strong to fading out they never where even a bit close to TH esters?

No. And I’m not saying everyone would get the same results, and I’m not trying to discourage anyone from experimenting. But in my experience if you brew (for example) a Wit and poured it into a pale ale, it would taste just like what you would expect, a wit poured into a pale ale. It wouldn’t produce a new exotic flavor that didn’t exist in either of the original beers. Which brings me back to my previous point - TH has a very distinct flavor, and it’s a flavor that’s strong enough to be able to compete with heavily hopped beers. In my opinion, if was as simple as 90% flavor of this style and 10% flavor of that style, someone would’ve figured it out by now. I could easily be wrong, but that’s my guess.

 

[beervoid]

No. And I’m not saying everyone would get the same results, and I’m not trying to discourage anyone from experimenting. But in my experience if you brew (for example) a Wit and poured it into a pale ale, it would taste just like what you would expect, a wit poured into a pale ale. It wouldn’t produce a new exotic flavor that didn’t exist in either of the original beers. Which brings me back to my previous point - TH has a very distinct flavor, and it’s a flavor that’s strong enough to be able to compete with heavily hopped beers. In my opinion, if was as simple as 90% flavor of this style and 10% flavor of that style, someone would’ve figured it out by now. I could easily be wrong, but that’s my guess.

I havent had many TH but the ones I did really have a hefe/wheat beer smell to it.
I poured a bit of a belgium wit bier in my neipa and wasnt impressed, the lower abv probably makes it thinner then you want so I dont know if this is worth comparison.

 

[Lakeside_Brewer5]

I havent had many TH but the ones I did really have a hefe/wheat beer smell to it.
I poured a bit of a belgium wit bier in my neipa and wasnt impressed, the lower abv probably makes it thinner then you want so I dont know if this is worth comparison.

To me this is evidence that it’s not actually 2 different beer styles, but the same exact wort, mashed and sparged, then split into a main batch and small batch (or 1 main and 2 small batches with the right equipment) for each yeast to interact with the wort in its own way.

 

[echoALEia]

@echoALEia Ive asked this before somewhere in HBT with no response though. How do you know how many cells are in a packet of SO4 to know you are under pitching? Fermentis states is 6B cells per gram, but if you go by Mr. Malty standards, its 20B cells. So this is tremendous variation. What do you go by to know you are under pitching and what do you think your pitch rate actually is? Thanks.

Hey sorry! Didn’t see the post from before. I guess in the end I really don’t. I just go by recommended pitching rates in beersmith then pitch less than that. For this batch 137g was recommended. I pitched 90g. No rhyme or reason why. Again, sorry about no responding before

 

[echoALEia]

With shaking I would be worried with over extraction and getting too much yeast in suspension, but I think you can dump through the bottom right?
How much contact time do you give the hops?

Recently 24 - 36 hours then I chill and dump.

 

[echoALEia]

LOL. If you don’t want to share your recipe bud, don’t share your recipe. There’s been a lot of people who contributed to this thread, who shared beer, who shipped beer to be test, etc. You through out an idea that was also thrown out about 80 pages ago. Nothing wrong with that, but let’s not act like you’re the only one contributing.

This seems more process oriented than recipe oriented, in my opinion. He has laid out good/detailed ideas for a process.

 

[troxerX]

To me this is evidence that it’s not actually 2 different beer styles, but the same exact wort, mashed and sparged, then split into a main batch and small batch (or 1 main and 2 small batches with the right equipment) for each yeast to interact with the wort in its own way.

Gosh, wanted to ask, you read through the entire thread?, LOL I stopped by panel #13 and re-started at #90...

Affirmative, all options are on the table, either a wort that we split and ferment separately and then re-inject or multiple worts fermented separately and then blended. One of the reasons I lean on different worts is that it gives you the capability to maximize its properties for the yeast you will be pitching and also allows you to add layers of texture and complexity vs one wort with a fixed set of properties. It also allows you to add more layers of different hops without overhopping/overbittering, dilution may play a role in saturation but we have to figure that one out. What I can’t think of right now is that if aroma is impacted. If splitting a wort, it has to be one properly designed to add all the sensory just like @echoALEia did. TH has multiple beers and who knows all these options are part of their processes. If let’s say some of their beers are made with multiple worts using the same principles, I might think they can use that to their advantage by brewing a huge large base batch and then split into different blends with their own other smaller batches, people see blending as a limitation, whatever approach we take I see it as an opportunity.

 

[Clyde McCoy]

Summary of findings:

The primary yeast in Julius, jjjuliusss, and Very Green matches the electrophoretic banding profile of Fermentis S-04 (as identified by @isomerization):

In fact, S-04 was the ONLY strain I could find until @Northern_Brewer suggested I try WLN agar. This is what various Tree House beers look like on WLN:

Everything on these plates tends to match the electrophoretic banding profile of Fermentis S-04, except for the large, beige colonies, which are 3-4 different strains:

(continued on next post, max 10 images)

 

[Clyde McCoy]

These strains are likely WB-06 and T-58 (as identified by @isomerization), but I’ll say these are unconfirmed because the profiles aren’t an exact match and may require a bit of PCR optimization to say more conclusively.

I’m again going to quote Chris White (White Labs) on mixed-strain fermentations:

“When yeast is pitched into beer, it starts to grow, entering into a log phase of growth after a few hours. This is when the bulk of the flavor compounds are produced. 12-36 hours into the fermentation. Therefore, if your goal is flavor, you need to add the multiple strains early on, preferably together. Note that if you just want another strain for bottling, or to complete attenuation, go ahead and add later. Very little flavor contribution happens here, unless the beer undergoes prolonged ageing”

You'll notice that on some of the WLN plates it looks like S-04 (small green colonies) is 99+% of the plate.

I'm pretty near my limit on the amount of time I can spend on this, but because the composition of yeast in the can will change over time (all beers above are typically analyzed at 1-2 months old) I am willing to test a few very fresh cans (ideally <10 days), PM me if you can mail. These will be the most indicative of the ratio of S-04 to everything else at the time of canning.

 

[echoALEia]

I’m so intrigued that Monkish uses 71b. I don’t even know where to begin with the questions for that. Is that first colony S33?

 

[Clyde McCoy]

I’m so intrigued that Monkish uses 71b. I don’t even know where to begin with the questions for that. Is that first colony S33?

Sorry to clarify, there is no match between Monkish and 71B (or between Tree House and 71B). It is somewhat similar to S-33, but would need to run them together. That Monkish profile might be Conan or LAIII, I haven't tested those.

 

[troxerX]

Summary of findings:

The primary yeast in Julius, jjjuliusss, and Very Green matches the electrophoretic banding profile of Fermentis S-04 (as identified by @isomerization):

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In fact, S-04 was the ONLY strain I could find until @Northern_Brewer suggested I try WLN agar. This is what various Tree House beers look like on WLN:


View attachment 699341View attachment 699342
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Everything on these plates tends to match the electrophoretic banding profile of Fermentis S-04, except for the large, beige colonies, which are 3-4 different strains:


View attachment 699347

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(continued on next post, max 10 images)

Just curious, do we know why these 3-4 yeasts look beige and S04 green? is this an artifact of the test itself or is there any biological reason?

 

[Noob_Brewer]

From what I understand about these agar plates is that the green is simply a dye in the agar and the yeast colonies take up the dye at different rates, so the presence of Visibly different colors of colonies points merely to the presence of multiple yeast strains.

I am unsure though if you can use thesetypes of qualitative analyses to come to a conclusion of the distribution of one type of yeast (all the small darker green colonies) vs another (larger beige colonies)

 

[Clyde McCoy]

Just curious, do we know why these 3-4 yeasts look beige and S04 green? is this an artifact of the test itself or is there any biological reason?

http://brettanomycesproject.com/2009/03/wln-agar-medium/

 

[Clyde McCoy]

From what I understand about these agar plates is that the green is simply a dye in the agar and the yeast colonies take up the dye at different rates, so the presence of Visibly different colors of colonies points merely to the presence of multiple yeast strains.

I am unsure though if you can use thesetypes of qualitative analyses to come to a conclusion of the distribution of one type of yeast (all the small darker green colonies) vs another (larger beige colonies)

You can if you've tested lots of colonies

 

[Noob_Brewer]

You can if you've tested lots of colonies

You've done a sh*t ton of work @Clyde McCoy as well as @isomerization has as well. Pretty cool watching all of this as a long time lurker in this thread. BTW....im on to you!

I'm pretty near my limit on the amount of time I can spend on this, but because the composition of yeast in the can will change over time (all beers above are typically analyzed at 1-2 months old) I am willing to test a few very fresh cans (ideally <10 days), PM me if you can mail. These will be the most indicative of the ratio of S-04 to everything else at the time of canning.

This is code for - give me FRESH beer and I'll continue with this escapade! LMFAO lol.

 

[Noob_Brewer]

seriously though, appreciate your work on this! This isn't easy and is time intensive!

 

[Clyde McCoy]

You've done a sh*t ton of work @Clyde McCoy as well as @isomerization has as well. Pretty cool watching all of this as a long time lurker in this thread. BTW....im on to you!


This is code for - give me FRESH beer and I'll continue with this escapade! LMFAO lol.

I use to think that until I realized how many hours I've put into this...

 

[isomerization]

I use to think that until I realized how many hours I've put into this...

It escalates quickly!

 

[Clyde McCoy]

It escalates quickly!

I think I crossed the 100 colony mark today...

It's looking like I will identify the same minor strains as you previously found (T-58 for sure, I'm still not 100% on WB-06), but I think they might be more rare than previously thought (going by WLN here and previous testing without WLN which was a needle in a haystack problem).

 

[isomerization]

It’s very interesting to me that the 3 other strains are metabolizing the dye. I could never find much info on WLN agar mechanics, but Chad Yackobson suggested Wit yeast were among Sacc spp that could do this, don’t know if that’s helpful or not though

 

[isomerization]

I think I crossed the 100 colony mark today...

It's looking like I will identify the same minor strains as you previously found (T-58 for sure, I'm still not 100% on WB-06), but I think they might be more rare than previously thought (going by WLN here and previous testing without WLN which was a needle in a haystack problem).

That’s cool in its own right, as I could have envisioned them having to change with the process scaling up. Speaks to how difficult their process is, given the quality issues reported.