[gimmeales]

I've got a bottle of Fantome Pissenlit I'd like to try culturing the yeast from.  In the past I've successfully washed and re-used yeast for multiple generations from fresh cultures, so I'm familiar with the process and sanitation measures that need to be taken.   Still, a couple questions though:

Isn't yeast from a bottled beer (especially an 8% example) pretty stressed and in bad shape by the time you drink it to warrant not culturing it up?  Especially considering an imported beer has endured a potentially many-weeks-long journey in getting to my fridge in potentially compromising environments (heat, light, etc)?

I know it can and has been done, but I'm wondering what I can realistically expect both in flavor profile and performance from culturing up from bottle dregs.  Also, any simple steps I can take to ensure they're as healthy as possible? (nutrients, oxygen, etc)?
Thanks!

[narvin]

It's hit or miss.  I've had great results from some beers (Saison Dupont, Delerium Tremens), and got nothing from others (Rochefort .  It's pretty obvious if the starter takes off and smells good.

The Fantome probably has some wild yeast or bacteria in there, so making a starter will change the proportions (and probably favor the sacc yeast).

[weithman5]

a new york brewery makes some beer for costco under the kirkland name.  their hefeweizen is not bad and i am thinking of bringing some of that up from the bottle.  it was the only beer in the pack that i liked and i don't normally like many wheat beers.  so i am going to try that yeast out if i can farm it

[gimmeales]

thanks Narvin, I'll definitely give it a try just because I haven't done it before.  With the variety of yeasts commercially available
in pure cultures, building up stressed yeast from a bottle never really made sense to me UNLESS there was some unique facet to a yeast (like the Fantome I have which may have some additional beasties in there).

I know Drew has made what sound like killer Saisons from Fantome bottle-dregs (at least I assume they were dregs), and Saison isn't a style that needs or wants a clean yeast, so figure it's as safe as any to try.

Others experiences and/or in-depth yeast knowledge still welcome!

Weithman, you might have some luck contacting the brewery that contract-brews for Costco, they might be willing to share and chances are they use a widely available commercial strain.  Even so, it might be process that makes the yeast perform a certain way, so just knowing the strain may not tell you everything.

[tschmidlin]

You should definitely do it.  If they make a beer that you think might have less stresses yeast of the same variety try culturing from that, but even still it's worth a try with what you have.

If you are really worried that the yeast is stressed, make a light starter, as little as 2% DME, so 2 grams in 100 ml of water.  You should definitely supplement with nutrients, the proportion will depend on what nutrient you're using.

Daughter cells from stressed mothers will themselves be weak, but the nicer the conditions the faster subsequent generations will recover.  The packaging and shipping conditions will have selected for cells that are best able to survive those conditions, but that doesn't mean that they will be so mutated as to be wildly different from the fermenting strain.

If you've made plates before, I recommend them for yeast culturing from bottles.  It is nice to just dump some dregs on the plate and look for what grows.  From there you can streak to other plates and pick a nice looking colony.  A plate recipe is the same as your starter recipe, just with some agar added (available from morebeer.com).

[thcipriani]

tschmidlin, your recommendation of culturing up in a starter using 2g/100mL is interesting. I just had an email discussion with Chris White about ideal yeast propagation and he said, "To really eliminate the crabtree effect, you need to be down under 1.010" which would be about the gravity of the wort you're suggesting. Not to be a thread hijacker but - do you have any experience with yeast that you've cultured with this gravity starter? What has your experience been with cell count and health?

[tschmidlin]

I'm a grad student getting a PhD in Biochemistry and have done a lot of work with yeast.  Our standard media is 2% glucose.  I propagate all of my fermenting strains with the same formula and have had great success growing stuff out of bottles.

Unfortunately though, between grad school and kids my brewing schedule is chaotic and I never know very far in advance when or what I'll be brewing, so I usually don't have time to pull something from the -80 and grow up a pitchable quantity.  And if I do have time for that, I normally don't have time to do cell counts or stain to check viability.

But I will say this; 2% glucose is research standard growth media for a variety of reasons.  One is the suppression of the crabtree effect, which will help the yeast stay healthy - face it, ethanol is bad for them.  Another is that glucose is easy to ferment, and if you're just trying to grow mass you want it to be easy on them.

That being said, I always use DME for my starters, never glucose.  You want the yeast to already have made and be making maltase when you throw them into your beer, it decreases the lag time.  Some people say that yeast grown in glucose lose the ability to ferment maltose, which is really incredibly unlikely unless you do it for a long time.  But they will take longer to get going, because glucose represses the expression of maltase so first they have to switch the maltase genes on.  So always feed your yeast maltose before putting them in your wort.

[thcipriani]

I'm convinced. I think I'll try 2% DME in my next batch of starter wort. Currently I've been doing starter wort at about 1.020 (that's actually what I emailed Dr. White about in the first place) and I haven't noticed any problems with yeast health or counts (however, I don't have a microscope [yet] so my "counts" are based on sedimented volume rather than counting in an actual hemocytometer).

Sorry to thread-jack gimmeales - FWIW I was listening to the Brew Strong on Yeast Rinsing the other day and towards the end Jamil was talking about growing up Dutchess from the bottle and he mentioned that he was able to retain most of the original character and balance of the Dutchess critters - I imagine that Fantome would probably behave in the same way.

Also, and take this for what it's worth (which may not be much), I've had success with the Wyeast BC Nutrient - it's zinc content may be less bio-available than servomyces, but it also contains other nutrients that the yeast can use.

[gimmeales]

Sorry to thread-jack gimmeales - FWIW I was listening to the Brew Strong on Yeast Rinsing the other day and towards the end Jamil was talking about growing up Dutchess from the bottle and he mentioned that he was able to retain most of the original character and balance of the Dutchess critters - I imagine that Fantome would probably behave in the same way.

Also, and take this for what it's worth (which may not be much), I've had success with the Wyeast BC Nutrient - it's zinc content may be less bio-available than servomyces, but it also contains other nutrients that the yeast can use.

Seems totally relevant to the topic at hand - I've learned alot and am now more confident to try this!  Thanks all!

[weithman5]

Weithman, you might have some luck contacting the brewery that contract-brews for Costco,

Thanks, i may give that a try.  depends on which is more lazy of me, calling or culturing

[dak0415]

I'm a grad student getting a PhD in Biochemistry and have done a lot of work with yeast.  Our standard media is 2% glucose.  I propagate all of my fermenting strains with the same formula and have had great success growing stuff out of bottles.

That being said, I always use DME for my starters, never glucose.  You want the yeast to already have made and be making maltase when you throw them into your beer, it decreases the lag time.  Some people say that yeast grown in glucose lose the ability to ferment maltose, which is really incredibly unlikely unless you do it for a long time.  But they will take longer to get going, because glucose represses the expression of maltase so first they have to switch the maltase genes on.  So always feed your yeast maltose before putting them in your wort.

Tom,
What do you think about doing the first and second step ups for a big starter with a glucose/dme mix and then the final starter with all DME?  I have been considering ways to reduce the cost of making large starters, short of doing a mash just for starter wort.

[tschmidlin]

Tom,
What do you think about doing the first and second step ups for a big starter with a glucose/dme mix and then the final starter with all DME?  I have been considering ways to reduce the cost of making large starters, short of doing a mash just for starter wort.

That will be totally fine, although I'm not sure you even need the mix.

Basically, high levels of glucose repress the expression of maltase regardless of the presence of maltose, and it happens quickly - in less than 20 minutes IIRC, the levels of maltase are falling.  The yeast will preferentially ferment the glucose, then when that drops below some threshold they will turn on the maltase genes and go after the maltose.  So by doing a mix for the first couple of steps they'll essentially be yo-yoing between the two sugars.  This is not especially stressful for the yeast though, they're fine - the only difference is the expression of all of the maltase genes.  I don't remember how many there are in the cascade . . . ok, according to SGD there are 10 that are directly involved in maltase, but only 2 are the actual enzymes themselves.  But that doesn't include all of the genes needed, because there are maltose permease genes.  Anyway, I wouldn't worry about these details much, I just thought you'd be interested.

Bottom line, supplementing your starters with glucose is fine, especially if the last step before pitching is all DME.  You could use sucrose (table sugar) for that matter, maltase can invert that too, it's cheaper than glucose, and won't suppress maltase the way straight glucose does.

[dak0415]

That will be totally fine, although I'm not sure you even need the mix.

The mix was more for yeast nutrients but I guess you have to add nutrients to the growth medium (distinguished from starter) the same way you have to add nutrients to mead.  What concentration would you recommend to grow the little buggers, and would you need more steps than using 1.040 wort?

[tschmidlin]

That will be totally fine, although I'm not sure you even need the mix.

The mix was more for yeast nutrients but I guess you have to add nutrients to the growth medium (distinguished from starter) the same way you have to add nutrients to mead.  What concentration would you recommend to grow the little buggers, and would you need more steps than using 1.040 wort?

I haven't tested it, but you might not actually need more.  When you encourage them to respire as opposed to ferment they get a lot more energy per gram of sugar, it's 19 times more efficient on paper.  If you leave them in the ethanol though, they will begin to use that as a carbon source provided there is still a source of oxygen (as there would be in a stirred starter).  So given enough time, 1.040 starters will yield more yeast.  But if you're pulling it off the 1.040 starter when there is still a lot of alcohol in the starter, then it might actually be better with the 1.010 starter.

I'd have to do some experiments to see when they cross over in terms of time/gravity/pitching rate to optimize growth.  It's probably known though, just not by me.

[dak0415]

So given enough time, 1.040 starters will yield more yeast.  But if you're pulling it off the 1.040 starter when there is still a lot of alcohol in the starter, then it might actually be better with the 1.010 starter.

What kind of time are we talking about, extra day, 3?