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

[couchsending]

Welp less than 12 hours later the isolates “labeled” T-58 and WB-06 are fermenting away in 40ml of 1.020 wort...

the one labeled S04... not so much.

 

[suregork]

I find it pretty funny that the only common strain identified between this thread and the reddit one is (apparently) WB-06!

The odds of yeast floating around in the air with matching patterns for the commercial strains is extremely low. I started with the TH beers too, so that would minimize that chance of cross contamination.

Is it possible that STA1 expression could be down-regulated if the yeast is introduced to a low sugar environment (eg like at dry hopping)? @suregork any thoughts here?

Finally, can anyone explain how I can update the first post? I’d like to repost the lost images of the PCR reactions there.

WB-06 is STA1+, but it has a deletion in the STA1 promoter. This deletion is common in many of the STA1+ strains (which is probably why many went unnoticed for such a long time). This significantly reduces the activity of STA1 (see our recent paper: https://link.springer.com/article/10.1007/s00253-019-10021-y ). STA1 expression is e.g. repressed by glucose. Anaerobic conditions also limit expression in the strains with the promoter deletion. WB-06 is very closely related to the Duvel strains from White Labs and Wyeast, as well as WLP644.

Edit: and in case my response on the end of the previous page gets missed, I'll quote it here just in case:

If someone has the TH isolates left, and an extra 125 USD per strain, then I recommend the whole genome sequencing route. MiGS does 625 Mb Illumina sequencing (50x coverage for yeast) for 125 USD.

https://www.migscenter.com/sequencing

I'd be happy to do the analysis and figure out the closest matches if someone orders the sequencing. The interdelta route will get you in the ballpark, but WGS if you want a more precise answer (provided the commercial strains have been sequenced as well; but very many have).

 

[Clyde McCoy]

If someone has the TH isolates left, and an extra 125 USD per strain, then I recommend the whole genome sequencing route. MiGS does 625 Mb Illumina sequencing (50x coverage for yeast) for 125 USD.

https://www.migscenter.com/sequencing

I'd be happy to do the analysis and figure out the closest matches if someone orders the sequencing. The interdelta route will get you in the ballpark, but WGS if you want a more precise answer (provided the commercial strains have been sequenced as well; but very many have).

Do you think two closely related strains could produce identical (or near identical) banding patterns?

 

[Northern_Brewer]

Do you think two closely related strains could produce identical (or near identical) banding patterns?

There's an example in isomerization's original PCRs, Windsor and London produce almost identical bands and are now known via genome sequencing to be very close relatives.

But it can be messy, fairly small underlying differences can make big differences to the perceived banding pattern (not least because humans are easily distracted by certain band patterns).

 

[JohnConnor]

Isnt it possible the bready thing comes from the malt bill?

Possible? I guess. I'd be more on board with someone saying malt bill amplifies it. But I'm with couch. They use either all S04 or some variant or it's a huge part of any mix they us. I can smell S04 from across the room. Yeasty/bready mix. The finished beer smell like it does when it's fermenting. That's why I stopped using it. I didn't enjoy that aroma at all. I just had fresh TH cans a month ago for the first time in a couple years. You get it the second you open the can. I forgot how much they smell like that. There are certainly much more approachable beers from an aroma standpoint. But their integration and mix of flavors.. that mouthfeel, light but substantial. Drying of the palate begs you to drink more. Second to none IMO.

 

[WIexile]

Have a interesting tidbit from my recent trip down to TG.

"David" the collab brew between TG and Tree House from a few years back was fermented using under pitched S-04 on Nate's persistence. This is one the reason's why TG now uses s-04 instead of 007 for many of their core beers.

 

[Clyde McCoy]

Have a interesting tidbit from my recent trip down to TG.

"David" the collab brew between TG and Tree House from a few years back was fermented using under pitched S-04 on Nate's persistence. This is one the reason's why TG now uses s-04 instead of 007 for many of their core beers.

<0.5 mil/ml/*plato?

 

[MrPowers]

Have a interesting tidbit from my recent trip down to TG.

"David" the collab brew between TG and Tree House from a few years back was fermented using under pitched S-04 on Nate's persistence. This is one the reason's why TG now uses s-04 instead of 007 for many of their core beers.

I'm mostly commenting so I start getting updates from this thread.

I did know that TG switched from 007 to 04 on most of their cores, I had not heard that a collab with Nate was one of the reasons they did that. They've been putting out some amazing beers since the new brewery went up.

 

[isomerization]

I'm mostly commenting so I start getting updates from this thread.

I did know that TG switched from 007 to 04 on most of their cores, I had not heard that a collab with Nate was one of the reasons they did that. They've been putting out some amazing beers since the new brewery went up.

Keep posting, but just wanted to share that you can click “Watch Thread” towards the top left of the page.

 

[brewpharm Hill]

Have a interesting tidbit from my recent trip down to TG.

"David" the collab brew between TG and Tree House from a few years back was fermented using under pitched S-04 on Nate's persistence. This is one the reason's why TG now uses s-04 instead of 007 for many of their core beers.

What is TG?

 

[couchsending]

Have a interesting tidbit from my recent trip down to TG.

"David" the collab brew between TG and Tree House from a few years back was fermented using under pitched S-04 on Nate's persistence. This is one the reason's why TG now uses s-04 instead of 007 for many of their core beers.

Its weird I don’t get any of the “bready” character from TG’s hoppy beers but they do finish a little acidic which is very SO4-like.

That being said I think TG might make some of the best (most shelf stable) modern IPAs you can buy. In a store or at a brewery. Recently had a four month old King Sue that was outstanding. I don’t think they get enough credit for their hoppy beers. They are my go to anywhere they distribute and as long as they’re kept cold they hold up so unbelievably well.

Sosus, Pompeii, King Sue, Pseudo Sue have all been great since they got the new brewery dialed in.

I know it’s long been thought that 007, 1098, and SO4 were all the same yeast but it’s interesting to see the yeast genome study and they’re really not similar at all. 1098 and S04 are somewhat related but they’re nowhere close to 007.

update: All three isolates survived and made their way into step 2 of 250ml 10* wort. Gonna step them up one more time and then test gravity and see how phenolic they are.

I’m no scientist. I have done everything I can possibly do to prevent any cross contamination. Spraying everything with alcohol, flaming everything, wearing gloves, working next to a flame, etc. however I don’t have a pressure cooker so wort is only boiled for 10 minutes, not sterilized.

 

[isomerization]

What is TG?

Toppling Goliath, brewery in Iowa.

 

[MrPowers]

What is TG?

Toppling Goliath.

That being said I think TG might make some of the best (most shelf stable) modern IPAs you can buy. In a store or at a brewery. Recently had a four month old King Sue that was outstanding. I don’t think they get enough credit for their hoppy beers. They are my go to anywhere they distribute and as long as they’re kept cold they hold up so unbelievably well.

Sosus, Pompeii, King Sue, Pseudo Sue have all been great since they got the new brewery dialed in.

I agree 100%. They have their packaging process down just like The Alchemist. If it's been kept cold, don't worry if it's 2-3 months old.

IIRC, they are using S-04 on their lower ABV cores (Pompeii, Sue, Gold Nugget, etc...) and 007 on their doubles still. That may have changed as they got the new brewery dialed in.

 

[isomerization]

Its weird I don’t get any of the “bready” character from TG’s hoppy beers but they do finish a little acidic which is very SO4-like.

That being said I think TG might make some of the best (most shelf stable) modern IPAs you can buy. In a store or at a brewery. Recently had a four month old King Sue that was outstanding. I don’t think they get enough credit for their hoppy beers. They are my go to anywhere they distribute and as long as they’re kept cold they hold up so unbelievably well.

Sosus, Pompeii, King Sue, Pseudo Sue have all been great since they got the new brewery dialed in.

I know it’s long been thought that 007, 1098, and SO4 were all the same yeast but it’s interesting to see the yeast genome study and they’re really not similar at all. 1098 and S04 are somewhat related but they’re nowhere close to 007.

update: All three isolates survived and made their way into step 2 of 250ml 10* wort. Gonna step them up one more time and then test gravity and see how phenolic they are.

I’m no scientist. I have done everything I can possibly do to prevent any cross contamination. Spraying everything with alcohol, flaming everything, wearing gloves, working next to a flame, etc. however I don’t have a pressure cooker so wort is only boiled for 10 minutes, not sterilized.

Totally agree on TG as well, only beer I don’t care for from them is Morning Latte (bitter in a bad way and thin).

Just like making a starter, I am a bit shocked that they grew so well though! Should have pretty distinct aromas from my recollection (I think I posted early in the thread about them).

 

[couchsending]

Totally agree on TG as well, only beer I don’t care for from them is Morning Latte (bitter in a bad way and thin).

Just like making a starter, I am a bit shocked that they grew so well though! Should have pretty distinct aromas from my recollection (I think I posted early in the thread about them).

I degassed a Morning Latte I think. FG was definitely north of 1.040 which surprised me.

 

[isomerization]

I degassed a Morning Latte I think. FG was definitely north of 1.040 which surprised me.

I will have to try another one sometime, must have been something with the coffee I didn’t like. Acrid bitterness can make the beer taste more thin than expected, but I would have guess in the low 20s or high teens, not above 1.040!

 

[MrPowers]

A question/theory that I have (it may have been brought up earlier) about Tree House's process is, assuming they are using 3 different yeast strains, what if they are:

- fermenting primary all the way out with S-04
- soft crashing the S-04 out/dropping the yeast
- warming back up, adding dry hops, adding WB-06/T-58, and possibly some amount of additional sugar

They could be using hop creep from the dry hops and some additional amount of sugar to get a controlled amount of hop/yeast interaction (biotransformation). It seems like that would be a much more predictable/controlled method than just pitching the WB-06/T-58 earlier in fermentation.

Edit: and that would also allow them to repitch the S-04 and help keep their costs down.

 

[beervoid]

A question/theory that I have (it may have been brought up earlier) about Tree House's process is, assuming they are using 3 different yeast strains, what if they are:

- fermenting primary all the way out with S-04
- soft crashing the S-04 out/dropping the yeast
- warming back up, adding dry hops, adding WB-06/T-58, and possibly some amount of additional sugar

They could be using hop creep from the dry hops and some additional amount of sugar to get a controlled amount of hop/yeast interaction (biotransformation). It seems like that would be a much more predictable/controlled method than just pitching the WB-06/T-58 earlier in fermentation.

Edit: and that would also allow them to repitch the S-04 and help keep their costs down.

I find it hard to believe that with this technique those yeasts would ferment out healthy without causing any off flavors as there is not much nutrient left in the wort at that point.
They might be pitching yeast nutrient with it?

 

[Taket_al_Tauro]

I find it hard to believe that with this technique those yeasts would ferment out healthy without causing any off flavors as there is not much nutrient left in the wort at that point.
They might be pitching yeast nutrient with it?

If that was the case then also bottle conditioning would produce off flavors.

 

[couchsending]

I find it hard to believe that with this technique those yeasts would ferment out healthy without causing any off flavors as there is not much nutrient left in the wort at that point.
They might be pitching yeast nutrient with it?

People bottle condition all the time with T-58 and it doesn’t produce off flavors.

I too support the idea that these yeasts aren’t pitched at the beginning of fermentation.

The yeast that was marked T-58 is definitely super phenolic and it sediments well.

still waiting to pass judgement on the other two after the next step up. The WB-06 doesn’t “smell” very phenolic I will say that.

 

[stickyfinger]

i just did an IPA using the 92/5/3 at pitching of so4/t58/wb06. the beer tastes great and has a sort of funky flavor, phenols, not any bubblegum though. i recently had a commercial DIPA and then one of mine right after and for some reason mine tasted like it had been smoked! it was so phenolic! 1) beers can taste very different in different sequences and 2) the t58 must have put the smokey character into the beer. it's very pleasant as long as i haven't had any other beer before it!

i was thinking about pitching so4 and a larger amount of wb06 next time. i usually get a lot of bubblegum or banana out of wheat beer strains. it might not be TH but it could be interesting.

 

[NJGeorge]

People bottle condition all the time with T-58 and it doesn’t produce off flavors.

I too support the idea that these yeasts aren’t pitched at the beginning of fermentation.

The yeast that was marked T-58 is definitely super phenolic and it sediments well.

still waiting to pass judgement on the other two after the next step up. The WB-06 doesn’t “smell” very phenolic I will say that.

Good stuff! Will you be brewing any beers with the isolates? Also, have you had luck replicating the smooth mouthfeel? Any tips you can share in regards to water adjustment and salt ratios?

 

[couchsending]

Good stuff! Will you be brewing any beers with the isolates? Also, have you had luck replicating the smooth mouthfeel? Any tips you can share in regards to water adjustment and salt ratios?

Doubtful honestly. I don’t really like the yeast profile of their beers. The next round of steps will be fermented in the low 60s without stirring just to see what the taste/smell is like. A little closer to a beer than what a starter would smell or taste like. The one labeled WB-06 intrigues me the most. If it’s interesting I might try something with it but doubtful.

I don’t think I can add anything that hasn’t been discussed ad nauseam already in this thread.

Their beers are really high in Potassium which I find odd. Obviously a lot of fruit juice is high in potassium as well. Maybe they’re adding something containing a lot of potassium. I’ve tried some KCL and found it rather off putting.

Definitely naturally carbonation of some sort. Not sure if they use sugar or wort/gyle. Supposedly the mouthfeel is different??

I don’t think there’s one thing that will get you that mouthfeel. It’s the combination of a lot of little things.

I think their water is rather high in bicarbonate and they don’t use RO. The bicarbonate/Ca reaction will precipitate Ca so you end up with less Ca in the final beer but you get all the other ions. I think this might be key. Not sure you can replicate using RO water. Just a hunch though.

 

[beervoid]

Doubtful honestly. I don’t really like the yeast profile of their beers. The next round of steps will be fermented in the low 60s without stirring just to see what the taste/smell is like. A little closer to a beer than what a starter would smell or taste like. The one labeled WB-06 intrigues me the most. If it’s interesting I might try something with it but doubtful.

I don’t think I can add anything that hasn’t been discussed ad nauseam already in this thread.

Their beers are really high in Potassium which I find odd. Obviously a lot of fruit juice is high in potassium as well. Maybe they’re adding something containing a lot of potassium. I’ve tried some KCL and found it rather off putting.

Definitely naturally carbonation of some sort. Not sure if they use sugar or wort/gyle. Supposedly the mouthfeel is different??

I don’t think there’s one thing that will get you that mouthfeel. It’s the combination of a lot of little things.

I think their water is rather high in bicarbonate and they don’t use RO. The bicarbonate/Ca reaction will precipitate Ca so you end up with less Ca in the final beer but you get all the other ions. I think this might be key. Not sure you can replicate using RO water. Just a hunch though.

How high with KCL did you go? And what kind of off putting smell taste did you get?

The ballast point study on mineral profiles showed that a starting profile with Ca of 80ppm dropped to around 34ppm final beer. 120ppm dropped to 40 so not far off.
Julius measured 64. Alter Ego 34.

I understand you need to ferment wb06 high to get the bubblegum banana out.

 

[beervoid]

If that was the case then also bottle conditioning would produce off flavors.

Good point so they would add just enough to be able to carbonate condition.

 

[Northern_Brewer]

The yeast that was marked T-58 is definitely super phenolic and it sediments well.

Hmm - I've not got my notes to hand but that doesn't sound like the T-58 I know, it's not strongly phenolic - bit of pepper but that's all, and drops well but doesn't flocc well from memory.

i was thinking about pitching so4 and a larger amount of wb06 next time. i usually get a lot of bubblegum or banana out of wheat beer strains.

Except WB-06 isn't a true hefe strain, despite the marketing name it's a weird member of the saison family that's most closely related to strains alleged to come from Duvel. And it's a biotransformation beast IME, it completely trashed hop flavour in a Chinook beer I made with it, whereas T-58 reduces hop flavour a bit, but in return gives you a lot more complexity, turning the typical Chinook grapefruit into more lime.

 

[couchsending]

Hmm - I've not got my notes to hand but that doesn't sound like the T-58 I know, it's not strongly phenolic - bit of pepper but that's all, and drops well but doesn't flocc well from memory.



Except WB-06 isn't a true hefe strain, despite the marketing name it's a weird member of the saison family that's most closely related to strains alleged to come from Duvel. And it's a biotransformation beast IME, it completely trashed hop flavour in a Chinook beer I made with it, whereas T-58 reduces hop flavour a bit, but in return gives you a lot more complexity, turning the typical Chinook grapefruit into more lime.

Fermentis lists flocculation as “-“ but sedimentation as “medium”. I’d say that what I’m seeing especially compared to what’s labeled “WB-06”.

I find T-58 very phenolic.

 

[savylemons]

Doubtful honestly. I don’t really like the yeast profile of their beers. The next round of steps will be fermented in the low 60s without stirring just to see what the taste/smell is like. A little closer to a beer than what a starter would smell or taste like. The one labeled WB-06 intrigues me the most. If it’s interesting I might try something with it but doubtful.

I don’t think I can add anything that hasn’t been discussed ad nauseam already in this thread.

Their beers are really high in Potassium which I find odd. Obviously a lot of fruit juice is high in potassium as well. Maybe they’re adding something containing a lot of potassium. I’ve tried some KCL and found it rather off putting.

Definitely naturally carbonation of some sort. Not sure if they use sugar or wort/gyle. Supposedly the mouthfeel is different??

I don’t think there’s one thing that will get you that mouthfeel. It’s the combination of a lot of little things.

I think their water is rather high in bicarbonate and they don’t use RO. The bicarbonate/Ca reaction will precipitate Ca so you end up with less Ca in the final beer but you get all the other ions. I think this might be key. Not sure you can replicate using RO water. Just a hunch though.

Fwiw I had a Ward Labs report done on a finished beer of mine brewed with a water profile built with RO consisting of nothing more than Na, CaCl2, and SO4 additions and it had near identical potassium levels (1177) to TH, so it's certainly possible to get those levels from the malt bill alone and no additional K supplementation.

 

[beervoid]

Fwiw I had a Ward Labs report done on a finished beer of mine brewed with a water profile built with RO consisting of nothing more than Na, CaCl2, and SO4 additions and it had near identical potassium levels (1177) to TH, so it's certainly possible to get those levels from the malt bill alone and no additional K supplementation.

What was the grainbill like? And how big was the beer?

 

[savylemons]

What was the grainbill like? And how big was the beer?

12 lbs Rahr 2 Row
2.5 lbs carafoam

1.068 OG

 

[couchsending]

That’s a lot of carafoam! I wonder if the Carafoam contributes to the higher levels of potassium? I’ve never had a hoppy beer tested with 1200ppm potassium. Some dark beers yes.

Made a 1.040 “saison” with Sterling and Centennial. Saved some wort and added it to the yeast from the previous step. Going to ferment these a little cooler and not on a stir plate.

Still not convinced what’s labeled WB-06 is actually that. The other stuff seems about right so far.

 

[isomerization]

wrong thread, sorry!

 

[beervoid]

12 lbs Rahr 2 Row
2.5 lbs carafoam

1.068 OG

Thanks. I got a similar grainbill planned for an experiment to see if high % of carapils/foam contribute the same amount to body as do oat or wheat. Any comments on this?

 

[savylemons]

Thanks. I got a similar grainbill planned for an experiment to see if high % of carapils/foam contribute the same amount to body as do oat or wheat. Any comments on this?

It was never a HUGE body booster for me. This grain bill and the finished beer water report was from 2 years ago and I've moved away from such high % carafoam since then. I ditched it completely for awhile since I was able to achieve long lasting meringue-like head and sticky lacing using only base malt when I addressed other things in my process. The potassium correlation speculation is interesting though, that never occured to me at all that they could be related. Now I wish I had a finished beer report of an IPA without carafoam to compare....

I know Nate loves to tout his "simple grain bill of 2-row and dextrin/carafoam" for many of his IPA's, I just never saw much benefit to carafoam personally, but I guess it's possible it may be doing something other than it's intended purpose of increasing foam stability.

 

[beervoid]

I had an Equilibrium Fractal Koru Set triple ipa. The mouthfeel was fantastic. They state to use only 2-row.

 

[NJGeorge]

I had an Equilibrium Fractal Koru Set triple ipa. The mouthfeel was fantastic. They state to use only 2-row.

Supposedly they use RO water and go chloride heavy. Also I see a lot of their beers just list wheat and oats as a base.

 

[NJGeorge]

That’s a lot of carafoam! I wonder if the Carafoam contributes to the higher levels of potassium? I’ve never had a hoppy beer tested with 1200ppm potassium. Some dark beers yes.

Made a 1.040 “saison” with Sterling and Centennial. Saved some wort and added it to the yeast from the previous step. Going to ferment these a little cooler and not on a stir plate.

Still not convinced what’s labeled WB-06 is actually that. The other stuff seems about right so far.

What about trying a small brew with the labeled WB-06 starter and see how a beer would be with some hops? Maybe it will be good. If it smells good in a starter, why not? How’s the SO4 starter?

 

[brewpharm Hill]

I haven’t had awesome results with carafoam either, but I like it more than carapils. I brewed Hoppy Thing by Nate recently and it had an amazing merengue like head and great mouthfeel with no high protein adjuncts or carafoam. Just 2 row, crystal 60, and honey malt. Used Brooklyn water which is pretty damn soft and treated my water with 5gm gypsum and 2.5 cacl. Surprisingly soft mouthfeel considering...good process and wort handling goes a long way

 

[brewpharm Hill]

Also plan on brewing my NEIPA this coming week once I get my Kolsch out of the fermenter. I'm think of testing S04 on its own to see how it compares to TH beers. I've never used it on its own. Out of curiosity, what would yall consider an underpitch of S04 in 5 gallons of 1.065 wort?

 

[couchsending]

So the next step up is done. Measured the gravities and pH of all 3. I left these in a ground level area in my house that’s around 62/63. No stir plate

OG 1.046 40ish IBUs Sterling Centennial

around 850ml of wort in each flask


“T-58”
- finished the fastest
- lowest gravity and pH
+ 1.008, 4.14ph
- definitely phenols but not strong
+ similar to Allagash’s yeast for White
- no “peppery” character
- the fast start is like T-58
- the low FG is not like T-58 typically
- I’d expect this one to finish the highest

“S04”
- 1.011
- 4.4 ph
- not 100% sure this is just S04
- this one took forever to get going
+ hence there maybe some wild yeast
- no “bready” character, not that tart
+ maybe because of 62* ambient??

“WB06”
- I would say this is not WB06
- highest FG at 1.0125
- pH 4.24
- no detectable phenols
- incredibly “bright” flavor
+ thought maybe the pH would be low
due to some “infection” but nope
Standard pH.

None of these results make me think what’s labeled as “wb06” is anything close to that which makes no sense. With what that PCR looks like I’d expect something that ferments lower and has phenols as just about everything genetically near to it is diastaticus and POF+ (Although wlp644 isn’t POF+??). Maybe the low temps have just slowed it down? The “starter” has cleared and the yeast has flocced

I am now tempted to brew something with what’s labeled as WB06 however I’ve got a bunch of other stuff planned at the moment. We’ll see.