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

[beervoid]

It says utilizing the process developed for juice project, thankful is aggressively cold hopped. Not that they’re going to share but I’d be curious at what temp and for how long.

Probably trying out the new theory that is floating around the internet.
BSG did a summary post on it. Also Janish wrote a piece on it.
https://bsgcraftbrewing.com/reevaluating-dry-hop-techniques

 

[Bizkit]

Pretty sure you’ve got it backwards. If it is S04 that they’re using they would be fermenting at a lower temp to begin with then raising towards the end at which point they’d be adding hops and capping to naturally carbonate the beers. Yes hop creep can cause diacetyl but if you’re adding the hops before or at the same time as a diacetyl rest you just need to wait a few extra days until the beer is negative for it before you cool. Yes they are probably using ALDC as are most commercial hoppy beer breweries by now.

On a side note I did just have one of the more enjoyable Tree House beers I’ve had in a while... Juice Project, Citra + Amarillo. They only add a small amount of extract in the kettle, no WP additions, and then one huge dry hop at cold temps for a short time. It could be a bit more bitter but it didn’t have that overpowering bready yeast ester that they all tend to have and you could actually pick out the hops.

It has been a while since Monson but i made note of the temp changes in that order i am 99% sure that is what i saw. could be wrong tho .... i mostly came here to push that there is a possibility they are creating their signature mouth feel, nose and flavor thru enzymes or processes in combination with S-04. I've personally experienced S04 coming very close especially when under pitched and capped... but there is something more at play.

 

[HopsAreGood]

Probably trying out the new theory that is floating around the internet.
BSG did a summary post on it. Also Janish wrote a piece on it.
https://bsgcraftbrewing.com/reevaluating-dry-hop-techniques

Haha, I shared that BSG link in the never ending NEIPA thread. I’ve been curious about dry hopping cold and currently have one dry hopping at 42 degrees.

 

[couchsending]

It says utilizing the process developed for juice project, thankful is aggressively cold hopped. Not that they’re going to share but I’d be curious at what temp and for how long.

Based on all the recent talk around cold hopping I’d say it’s pretty darn cold. Sub 40 probably

http://scottjanish.com/a-case-for-short-and-cool-dry-hopping/

 

[beervoid]

On that note, did any people here try out lower time dryhopping?
I did a 14c and 1 day and did not like the profile at all. Very greenish

 

[beervoid]

Haha, I shared that BSG link in the never ending NEIPA thread. I’ve been curious about dry hopping cold and currently have one dry hopping at 42 degrees.

Interesting. How long contact are you aiming for? I got to test more but first results where mediocre. Keep us updated if u can.

 

[HopsAreGood]

Interesting. How long contact are you aiming for? I got to test more but first results where mediocre. Keep us updated if u can.

2-3 days. I’ll let you know how it goes.

 

[couchsending]

I think there are a lot of variables at play here when it comes to cold dry hopping.

First and Foremost the quality of your hops. Cant stress enough how much better the hops are that TH has than what we as homebrewers buy.

What was the hopping load in the kettle? What was the pH of the beer when you dry hopped? What varieties of hops did you use? All things to be aware of when evaluating this process.

 

[TBryerton]

How do you know it’s one huge dry hop at cold temps for a short time? Not saying you’re wrong, just curious how you know that. The only info I can find is in the description on their website, and it doesn’t go into quite that level of detail.

I’m fairly certain Nate has stated in the past that he adds all DHs at one time. Obviously that process could change as their beers evolve.

 

[NJGeorge]

I think this is certainly something we should pursue based on the results we've had. They're absolutely capping fermenters at some point, most likely by day 7. They believe in biotransformation in their core beers. Also, after taking a look at this photo they shared on their website photo stream, its clear theyre both dry hopping and cold crashing BEFORE going to the brite tank. The caption states this beer is super treat on its way to the brite, and the condensation on the outside tells us its been crashed already. Obviously, that color can only come after dry hopping as well.

There may be some enzyme that 1) helps with mouthfeel 2) helps with biotransformation or 3) shuts down diastaticus yeast

View attachment 707007

When I chill my conical to around 50-60F this happens during warmer months. Could be anything.

 

[NJGeorge]

It has been a while since Monson but i made note of the temp changes in that order i am 99% sure that is what i saw. could be wrong tho .... i mostly came here to push that there is a possibility they are creating their signature mouth feel, nose and flavor thru enzymes or processes in combination with S-04. I've personally experienced S04 coming very close especially when under pitched and capped... but there is something more at play.

I've said this before, but what about using amino acids to increase esters? Valine and or leucine. There are extensive papers written on that stuff. I tried leucine once with t-58, 33 and 256 with some strong banana, however, ferm temp was too warm and I got some fusels.

 

[chieftain]

Pretty sure you’ve got it backwards. If it is S04 that they’re using they would be fermenting at a lower temp to begin with then raising towards the end at which point they’d be adding hops and capping to naturally carbonate the beers. Yes hop creep can cause diacetyl but if you’re adding the hops before or at the same time as a diacetyl rest you just need to wait a few extra days until the beer is negative for it before you cool. Yes they are probably using ALDC as are most commercial hoppy beer breweries by now.

On a side note I did just have one of the more enjoyable Tree House beers I’ve had in a while... Juice Project, Citra + Amarillo. They only add a small amount of extract in the kettle, no WP additions, and then one huge dry hop at cold temps for a short time. It could be a bit more bitter but it didn’t have that overpowering bready yeast ester that they all tend to have and you could actually pick out the hops.

So my most recent effort, with underpitched S04 (~150 ml of slurry), fermented at 62 degrees for first few days, was my best beer in quite a while. OG around 1.065. Have had oxidation issues lately so I dry hopped in my conical around 1.020 and raised temp to 66. It finished around 1.014. Primarily Galaxy with some Sabro. The finished product comes across with grapefruit, peach, and citrus.

 

[beervoid]

Did a batch with s-33 and have to say the esters are very similar to s-04, but more stronger and in the beginning it was a bit more fruity instead of belgiumy bubblegum.
After some time it completely overwhelms my beer. Blending it with some london ale iii didnt make it much better.
Still prefer la3 way over all the other yeast experiments so far.
Allthough I do identify the bubblegum in both yeasts as to be part of the tree house ester profile, in their beers its definitely very subtle, I cant see how you can control this without blending honestly. I dont get the juice factors with these yeasts and they cover up alot of hop flavor.

 

[NJGeorge]

Has anyone on this thread or on HBT successfully used a Belgian yeast or any yeast to only produce bubblegum or banana? What about peppery and not clovey?

 

[Northern_Brewer]

It's OT, but Omega have a new yeast, OYL-400 Bananza Ale, which AIUI is a hefe yeast they've knocked out the POF on so it's guaranteed pure banana and no clove. Obviously not what Nate is using, but may be of interest.

 

[NJGeorge]

It's OT, but Omega have a new yeast, OYL-400 Bananza Ale, which AIUI is a hefe yeast they've knocked out the POF on so it's guaranteed pure banana and no clove. Obviously not what Nate is using, but may be of interest.

Could use this yeast to get closer in terms of replicating their house taste. Possible blend of split batch with another yeast.

Pretty cool that they can remove the POF gene. Wonder if it’s possible to do this with other yeasts at home. Can they mutate over time to get rid of that gene or does this have to be done on a lab with crazy equipment?

 

[suregork]

Could use this yeast to get closer in terms of replicating their house taste. Possible blend of split batch with another yeast.

Pretty cool that they can remove the POF gene. Wonder if it’s possible to do this with other yeasts at home. Can they mutate over time to get rid of that gene or does this have to be done on a lab with crazy equipment?

They've inactivated one of the genes required for the POF phenotype with CRISPR/Cas9 editing. In theory it could be done at home, but in practice it requires some lab equipment and experience (e.g. confirming a successful modification). Genetic engineering in yeast nowadays is super easy. Hoping the EU will allow commercial use of CRISPR-edited yeasts one day.

 

[isomerization]

They've inactivated one of the genes required for the POF phenotype with CRISPR/Cas9 editing. In theory it could be done at home, but in practice it requires some lab equipment and experience (e.g. confirming a successful modification). Genetic engineering in yeast nowadays is super easy. Hoping the EU will allow commercial use of CRISPR-edited yeasts one day.

Should be an easy phenotype to confirm without PCR

 

[Northern_Brewer]

They've inactivated one of the genes required for the POF phenotype with CRISPR/Cas9 editing.

It is CRISPR then? I did wonder, but hadn't seen anything definite. Must be one of the first for-sale CRISPRd products in any market?

Pretty cool that they can remove the POF gene. Wonder if it’s possible to do this with other yeasts at home. Can they mutate over time to get rid of that gene or does this have to be done on a lab with crazy equipment?

Well.... On one level, this is just kind of reversing what happened when the original hefe yeast was created, when a kolsch yeast naturally picked up a chunk of DNA from a saison yeast that included the POF cassette. So in theory you could grow a load of yeast in stressy, mutation-friendly conditions and eventually one of more of them would get a mutation in a POF gene. But that's the equivalent of giving a monkey the complete works of Shakespeare and expecting it to rip out the exact piece of paper that says "to be or not to be", whereas this is a super-targetted approach that says to someone with a scalpel "cut out the bit of paper on the third line of p731, from words 2 to 9"

 

[El Duderino]

So my most recent effort, with underpitched S04 (~150 ml of slurry), fermented at 62 degrees for first few days, was my best beer in quite a while. OG around 1.065. Have had oxidation issues lately so I dry hopped in my conical around 1.020 and raised temp to 66. It finished around 1.014. Primarily Galaxy with some Sabro. The finished product comes across with grapefruit, peach, and citrus.

That sounds like a great beer. Mind sharing the recipe/Hop Schedule?

 

[chieftain]

That sounds like a great beer. Mind sharing the recipe/Hop Schedule?

Sure thing. This netted 6 gallons into the fermenter, 5 gallons into the keg.

Water Profile
100% distilled water, aimed for 50 Ca, 16 Mg, 46 Na, 135 SO4, 104 Cl

Grist

12 lbs​	Pale Malt (2 Row) US (2.0 SRM)​	Grain​	6​	72.7 %​
3 lbs​	Pilsner (2 Row) Bel (1.2 SRM)​	Grain​	7​	18.2 %​
1 lbs​	White Wheat Malt (2.4 SRM)​	Grain​	8​	6.1 %​
8.0 oz​	Cara-Pils/Dextrine (2.0 SRM)​	Grain​	9​	3.0 %​

Hop Schedule
Boil

0.25 oz​	Columbus/Tomahawk/Zeus (CTZ) [15.50 %] - Boil 60.0 min​	Hop​	10​	11.0 IBUs​
1.00 oz​	Galaxy [14.00 %] - Boil 5.0 min​	Hop​	11​	7.9 IBUs​
0.50 oz​	Sabro [14.00 %] - Boil 5.0 min​	Hop​	12​	4.0 IBUs​

Steeped Hops

Amt​	Name​	Type​	#​	%/IBU​
3.00 oz​	Galaxy [14.00 %] - Steep/Whirlpool 30.0 min​	Hop​	13​	22.9 IBUs​
2.00 oz​	Sabro [14.00 %] - Steep/Whirlpool 30.0 min​	Hop​	14​	15.3 IBUs​

Dry Hop - 7oz Galaxy, 3 days

 

[Clyde McCoy]

Never really looked at this closely:

Nate's Hoppy Thing recipe says "Ferment at 66°F (19°C) and turn off the temperature control as fermentation vigor slows. Once desired gravity is reached, rest for a few days and dry hop for 4–6 days. Rack to a corny keg and force carbonate at 38°F (3°C) for two weeks."

Looks like they're fermenting at 66F, soft crashing to 60-62F for dry hopping, then cold crashing.

Still not sure if they co-pitch the minor strains with S-04 or add at dry hop. My guess is co-pitch, but who really knows other than their brewers.

Going to try co-pitch S-04 (11.5 g, 97%) & T-58 (0.35 g, 3%) on a future batch.

 

[echoALEia]

Do you think it’s possible they use hop cannons? The same company that made their panel makes them.

https://www.premierstainless.com/hop-cannon-yeast-brinks/

 

[Clyde McCoy]

Do you think it’s possible they use hop cannons? The same company that made their panel makes them.

https://www.premierstainless.com/hop-cannon-yeast-brinks/

No idea, but using a hop cannon rather than climbing on top of their FVs would make sense.

 

[couchsending]

Last I saw they still dry hop from the top with buckets of hops.

 

[NJGeorge]

No idea, but using a hop cannon rather than climbing on top of their FVs would make sense.

 

[couchsending]

They have catwalks between all the FVs (or did when I was last there). Why does it matter?

A lot of places have gone away from hop cannons or recirc devices cause the chance of someone not purging the device properly and introducing a bunch of o2 is much greater than just dropping them in through the top. Places with big enough FVs also generally have catwalks to make it safer or at least scissor lifts.

 

[beervoid]

Has anyone tried the Curiosity 104 or 105? It's claimed there they use a new yeast they never tried before. I've been looking through some reviews on Untappd and allthough people mention it's different it still has the typical juice/candy hop flavor.
Really makes me wonder how much of the signature Tree House flavor is yeast driven.

 

[TBryerton]

Has anyone tried the Curiosity 104 or 105? It's claimed there they use a new yeast they never tried before. I've been looking through some reviews on Untappd and allthough people mention it's different it still has the typical juice/candy hop flavor.
Really makes me wonder how much of the signature Tree House flavor is yeast driven.

The recent curiosities that don’t use the ‘house yeast’ don’t taste anything like a typical Treehouse IPA. Zero of that flavor you’re referring to. The first one reminded more of Heady Topper than a TH beer.

 

[NJGeorge]

The recent curiosities that don’t use the ‘house yeast’ don’t taste anything like a typical Treehouse IPA. Zero of that flavor you’re referring to. The first one reminded more of Heady Topper than a TH beer.

Agreed, I felt the same way. It also eventually cleared.

 

[beervoid]

The recent curiosities that don’t use the ‘house yeast’ don’t taste anything like a typical Treehouse IPA. Zero of that flavor you’re referring to. The first one reminded more of Heady Topper than a TH beer.

Agreed, I felt the same way. It also eventually cleared.

Sounds like vermont yeast to me.

 

[Andre3000]

Sure thing. This netted 6 gallons into the fermenter, 5 gallons into the keg.

Water Profile
100% distilled water, aimed for 50 Ca, 16 Mg, 46 Na, 135 SO4, 104 Cl

Grist

12 lbs​	Pale Malt (2 Row) US (2.0 SRM)​	Grain​	6​	72.7 %​
3 lbs​	Pilsner (2 Row) Bel (1.2 SRM)​	Grain​	7​	18.2 %​
1 lbs​	White Wheat Malt (2.4 SRM)​	Grain​	8​	6.1 %​
8.0 oz​	Cara-Pils/Dextrine (2.0 SRM)​	Grain​	9​	3.0 %​

Hop Schedule
Boil

0.25 oz​	Columbus/Tomahawk/Zeus (CTZ) [15.50 %] - Boil 60.0 min​	Hop​	10​	11.0 IBUs​
1.00 oz​	Galaxy [14.00 %] - Boil 5.0 min​	Hop​	11​	7.9 IBUs​
0.50 oz​	Sabro [14.00 %] - Boil 5.0 min​	Hop​	12​	4.0 IBUs​

Steeped Hops

Amt​	Name​	Type​	#​	%/IBU​
3.00 oz​	Galaxy [14.00 %] - Steep/Whirlpool 30.0 min​	Hop​	13​	22.9 IBUs​
2.00 oz​	Sabro [14.00 %] - Steep/Whirlpool 30.0 min​	Hop​	14​	15.3 IBUs​

Dry Hop - 7oz Galaxy, 3 days

This looks great man. Gonna try it!

 

[beervoid]

Was just pondering on this. How likely is it that the dregs harvested from cans are yeast that mated with each other and mutated? I recall someone made a vermont/london ale 3 hybrid yeast.
I think this was discussed before but what if we take a can of beer from which we are certain they use yeast A. And test this and compare it with yeast A thats commercially available.
What if its a yeast blend?
I'm wonder if once you start blending yeasts how sure is it the original culture survives?

 

[brewpharm Hill]

Was just pondering on this. How likely is it that the dregs harvested from cans are yeast that mated with each other and mutated? I recall someone made a vermont/london ale 3 hybrid yeast.
I think this was discussed before but what if we take a can of beer from which we are certain they use yeast A. And test this and compare it with yeast A thats commercially available.
What if its a yeast blend?
I'm wonder if once you start blending yeasts how sure is it the original culture survives?

I'm sure some of the microbio guys in here can confirm to deny... but given fermentation conditions I think it is highly unlikely that breeding took place between different strains in the cans.

When blending yeasts and repitching from a slurry a single strain will likely dominate in time in terms of population. To get those strains to sexually reproduce takes very specific conditions if I recall correctly. IF they were to sexually reproduce I suppose it would be feasible for a dominate strain to present itself that way.

@Clyde McCoy and @isomerization - What say you, maestros?

 

[suregork]

Yes, very unlikely that strains in a blend would cross-mate and a hybrid would take over the population in a can. Especially since we are talking about brewing yeast, which are quite challenging to breed with as they are sterile (i.e. do not form viable spores) and polyploid. However, if one were to blend say 10^9 cells of two parent strains in wort (e.g. S-04 and T-58 as they are constantly mentioned here), then maybe a few hybrids would form during the fermentation process. Isolating those hybrids is another challenge completely (needle in a haystack without good selection markers).

 

[Clyde McCoy]

Yes, very unlikely that strains in a blend would cross-mate and a hybrid would take over the population in a can. Especially since we are talking about brewing yeast, which are quite challenging to breed with as they are sterile (i.e. do not form viable spores) and polyploid. However, if one were to blend say 10^9 cells of two parent strains in wort (e.g. S-04 and T-58 as they are constantly mentioned here), then maybe a few hybrids would form during the fermentation process. Isolating those hybrids is another challenge completely (needle in a haystack without good selection markers).

Yep, fully agree. I also think "mutated" is a misused phrase by brewers (including professional brewers) who don't know anything about single nucleotide polymorphisms, short sequence insertions or deletions, recombination and gene conversion, short tandem duplications, gene and segmental duplications, transposable elements, gross chromosomal rearrangements, or ploidy changes etc. (sorry).

How do you prep your gDNA? For speed I have been near-boiling individual colonies in a thermocycler at 99C for 10’. I think this quick-and-dirty approach might be preventing the amplification of larger products. Extension and annealing changes haven't really made a difference (using GoTaq). Going to try Q5.

Have you tested anything other than the delta primers for strain-level identification?

 

[beervoid]

Ok thanks for clearing that up, I was just "typing out loud" here.

 

[TheHairyHop]

I’ve been doing a lot of research into anything Nate and co. have ever put out in terms of interviews, beer descriptions, pictures, etc. Take a look at the attached pictures (some have been posted here before, I’m sure) and chime in on if you agree with my lone of thinking. 1) his cellar setup circa 2012 with 3 Blichman 14 gallon fermenters also has 2 kegs turned fermenters, as evidenced by the blowoff hoses running out of them. Why buy/make these when you are already purchasing what was at the type the top of the line fermenters? This leads me to believe he needed an FV with the capability to pressurize while fermenting. If he changed the 4” TC cap he could pressurize while fermentation is completing (ie some small natural carbonation, for that soft mouthfeel). This leads me to number 2)... he has 2 flasks with starters going right next to the 2 keg-menters, almost like he’s preparing to pitch a 2nd yeast in each after fermentation has been going. This leads us towards pitching dry S04 first with no starter, then waiting until some/most of fermentation is done to pitch a starter of T58 and/or WB06 that’s already ready to go and capping the FV to finish fermenting. 3) the final variable here is when would dry hopping be performed? The biotransformation campy would say it’s pitched with the starter before the FV is capped. This would be similar to how @echoALEia arrived at his almost-there beer that’s been discussed. If capped, there should be no “blowing-off” of important oils and flavor/aroma compounds, as well. 4) Nate’s use of brite tanks is well known, so perhaps he’s fermenting in a typical FV until 5-10 points before final gravity, transferring to the brite, pitching the starter and dry hopping, and capping the beer for good. He’d add necessary CO2 to get the carbonation level needed and cold crash out the hop material before canning from the brite. This would explain some of the trub/hop residue in cans that people occasionally get.

Those starters could just be for krausening, and have nothing to do with a yeast blend. My setup sometimes looks similar to that picture, and I'm not doing anything fancy with my yeast

 

[beervoid]

Did some interdelta fingerprints (delta12/delta21 and Phusion polymerase) of some Escarpment and White Labs strains at work today. I'm not sure if they are of any help in IDing the strains . One observation is that the fingerprints of WB06, WLP570 and WLP644 are identical (and genome data suggests they are very close). The UK ale strains also have quite similar profiles, they can be quite difficult to tell apart on an agarose gel (these are run on a ZAG capillary electrophoresis device). The bands at 75bp and 20kbp are markers added to each sample. Clyde McCoy, what polymerase and extension time are you running? Taq and 30s? You barely have any bands above ~500-700 bp. If you extend to one minute you could better range.
View attachment 702224
I haven't followed this thread very closely, but it seems most of the focus has been on dry strains. Do we know they certainly use dry strains? Based on the fingerprints it could be WLP644 (POF- is the key here) and almost any UK ale strain. If one wants to stick to S-04 or similar, maybe a dark horse could be WLP540 (i.e. the Rochefort strain) which is closely related. Wasn't there some interview posted where Rochefort 8 was named as an "a-ha beer".

The aha beer and a high thiol releaser?
Maybe those Tree House esters are insane amounts of thiols?
When listening to this podcast I observed an interesting coincidence.
https://www.masterbrewerspodcast.com/195?t=0
If you look at the papers the yeasts that are high thiol releasers mentioned here.
https://my.ltb.io/www/#/
WLP540 is on the 4th highest place of high thiol releasing yeasts.

 

[couchsending]

The aha beer and a high thiol releaser?
Maybe those Tree House esters are insane amounts of thiols?
When listening to this podcast I observed an interesting coincidence.
https://www.masterbrewerspodcast.com/195?t=0
If you look at the papers the yeasts that are high thiol releasers mentioned here.
https://my.ltb.io/www/#/
WLP540 is on the 4th highest place of high thiol releasing yeasts.

So based on cross referencing the @suregork , Gallone et all, and that Thiol research the yeasts that I can decipher that medium or above thiol releasing capabilities are:

WLP410
WLp550
WLP540
Wyeast 1028

Be066 is in the Vermont Ale area as well but the Suregork tree and Gallone one is kinda different in that area so I’m not really sure.