Guide to Making a Frozen Yeast Bank

[zam216]

Thanks for the feedback appreciate it! I'm glad you liked the cereal choice

 

[sadowski25]

Hey,
I slowly reading this thread and a book " Yeast: The practical guide to beer fermentation" by Chris White and Jamil Zainasheff .
When I relocate my knifemaking workshop from Basement to garage, I have 2 clean rooms to use with steady 18*C in winter and summer.

In one I want to make very nice Yeast laboratory and bank.
In second home brewery.
Centrifuge, autoclave, laminar hood, -80*C freezer, 2*C - 45*C incubator with magnetic stirring plates ,microscope with mounted DSLR , fridge should be ok for that ? ( I know for home is overkill, but I like this type of precise work )

Should be a good lab for taking clean wild strains of yeast?
(And a make nice amount of micro-starters from yeast got from store )

 

[pricelessbrewing]

Super overkill for a personal bank.

Keep it simple, plating for isolation cultures and ensuring purity + frozen bank.


No need to spend all that money and equipment if it's just for you and maybe some friends. There's no way you will ever overcome that investment with personal or even homebrew club usage.

If it's a pro setup, then sure MAYBE.

 

[sadowski25]

But I will get that stuff from my Aunt
She is microbiologist, and she want to give me her old stuff, but full functional (Centrifuge, Autoclave, small laminar hood and freezer)
I will only need to buy good microscope and build mentioned incubator ( from fridge )

Hmm investment...
I already spend around 25.000$ on my knifemaking workshop ( but I'm a full-time knifemaker)

 

[pricelessbrewing]

Fair enough, sounds like you have a good setup coming up to you.

 

[butterpants]

I like the cut of you jib fella.

.... but that level of sophistication wouldn't have stirplates in an incubator.... it would be shakers.

Orbital shakers are better than stirplates... less shear on cells. At the homebrew growth scale I don't think it matters much cuz it's pretty easy to pitch high but as James Bond mentioned, shaken is always better than stirred.

 

[sadowski25]

Is possible to step-up Lactobacillus into bigger amount (clean strains - brevis etc. ) and make a 1,5ml micro-starters for banking ?
From 1 pack then I can make nice amount of starters with clean culture ( L. Brevis etc. are pricey here >.>)

From what I reading, with time and practice I'll be able to isolate even bretts from wild forage ?

 

[butterpants]

You can slant lacto. I use MRS agar. LAB grows like friken wildfire on this stuff.

I have no experience with freezing small amounts and reviving them. I'm sure someone here has.

Stepping up is easy and you need a suprisingly small amount of total growth media because bacteria are small and have a badass growth curve. Growing up in MRS media may not be optimal (well it WILL be for cell counts.... but it tastes n smells awful). You will want to propigate in a mixture of DME, Glucose, DAP and CaCO3.

http://sourbeerblog.com/lactobacillus-2-0-advanced-techniques-for-fast-souring-beer/

Dr Lambic has a big writeup on it. Super detailed

 

[jungbrasil]

: Lado a otro: :tanque:

 

[Breadontap]

I'm pretty sure the American Sours book says you can just feed your lacto and pedio with apple juice every couple of weeks and it will keep. No need to freeze or slant/plate. You can treat them like a sour dough starter. Just keep it fed.

 

[butterpants]

I'm pretty sure the American Sours book says you can just feed your lacto and pedio with apple juice every couple of weeks and it will keep. No need to freeze or slant/plate. You can treat them like a sour dough starter. Just keep it fed.

Problem with that is growth and longevity can be self-limiting based upon how low the pH goes.

 

[Breadontap]

Your right. It did mention adding chalk to keep the ph in check with consistent decanting and feeding.

 

[sadowski25]

From what labs do you suggest to first get Bretts and lacto ?

 

[Breadontap]

I'm a white labs guy myself but really who ever has the type of Brett or lacto you want. Look around the American sours book suggested http://www.eastcoastyeast.com/

I've never used them personally. It looks like they have a several blends.

I'm curious how often you plan to use lacto? I only use it when brewing a Berliner.

 

[butterpants]

Goodbelly Caps - L. plantarum

White Labs - L. brevis

All u need.... Brett... like a million good options

 

[pricelessbrewing]

FWIW white labs lacto is pretty well known to contain yeast, and so it's not a good option for sour worting or sour mashing. Combine that with their very low cell counts for lacto, and brett, and I don't recommend buying from them for those purposes.

If you can get it, omega lacto blend is amazing.

 

[butterpants]

If you're proping it yourself, cell counts don't matter... growth is exponential and rapid.

The Omega lacto blend is plantarum and brevis if I remember correctly... hence why I mentioned the two. Blend them yourself!

WL yeast has lacto in it too... I'm glad someone else noticed this QC issue.

 

[butterpants]

oh and if you propigate your LAB at a high enough temp the yeast don't survive (for the paranoid and people who can't isolate)

 

[pricelessbrewing]

oh and if you propigate your LAB at a high enough temp the yeast don't survive (for the paranoid and people who can't isolate)

I read someone tried this on the MTF group like two weeks ago, can't remember if it was successful or not. IIRC it was incubated at ~120-140?

 

[butterpants]

140 sounds pretty damm high. Not sure the LAB is gonna like it. Funk guys could have, I'm not aware of the experiment.

I think 120F is a nice place to start.

 

[BlackFurnaceBrewing]

Waiting for the ole HLT to come up to temp, so thought I would update on my yeast ranching experience. I have been freezing yeast since last november with phenomenal results. I have 8 different strains right now in a small dorm-sized freezer that is used just for yeast and hops. The yeast, two samples each, are stored in 45 ml autoclavable vials in a custom closed-cell foam locker I made just to protect my investment.

Here is how I stored the yeast:

- autoclaved several (two dozen or so) vials with 14 ml of glycerin and recommended dosage of vitamin c each and store for easy use later

- purchased new and healthy White Labs strain 1 month old or less

- Set up burners, sterilized counter and used disposable sterile pipette to split WL strain into two storage vials (approx 17 -20ml) of yeast slurry each

- Recorded strain and all pertinent info and stored vials in fridge for 48 hours in preparation for freezing

- Placed vials in foam locker and froze for later use

Here is how I re-animate:

- Pull strain from freezer and place in fridge ovenight
- Sanitize 1000ml flask and then put in 175ml of canned sterile 1.040 wort
- Warm yeast with hand, and then place in flask/wort
- Let sit for 1/2 hour and then place on heated stir plate for 24 - 30 hours
On second or third stage, pull one 20 ml yeast slurry sample and place in 45 ml storage vial. Follow freezing procedures above. Pull 3ml sample and follow yeast counting procedure (with microscope).

So far my starters have been incredibly healthy. I know it sounds crazy, but they seem even more lively than before I started freezing yeast. Great/healthy/tasty batches of brew as well. Needless to say, after 8 successful starters, I am now a complete convert to yeast ranching.

 

[butterpants]

Nice writeup!

Heated stirplates worry me.... unless you've got a brand new, digitally controlled one. Some of the surplus ones (Corning especially) are known to loose heat control after prolonged usage. What happens is they go on full high. This is really hot and could burn down your house. It's not enough to melt pyrex but it will melt tinfoil. Just google some of the horror stories.

I stopped using a heated stirplate for that reason.... remote as it is. I like my house. Got a SCREAMING deal on ebay for a person who didn't know what they had
... now I use this for small heated n shaken starters off slants and don't heat my larger volume step ups

 

[butterpants]

here it is

View attachment 1463930783204.jpg

 

[BlackFurnaceBrewing]

Nice writeup!

Heated stirplates worry me.... unless you've got a brand new, digitally controlled one. Some of the surplus ones (Corning especially) are known to loose heat control after prolonged usage. What happens is they go on full high. This is really hot and could burn down your house. It's not enough to melt pyrex but it will melt tinfoil. Just google some of the horror stories.

I stopped using a heated stirplate for that reason.... remote as it is. I like my house. Got a SCREAMING deal on ebay for a person who didn't know what they had
... now I use this for small heated n shaken starters off slants and don't heat my larger volume step ups

That is a nice get off of Ebay!

All my fermentation, including starters, takes place in my basement which stays 52 to 57F depending on time of year. A heated stir plate is a must for me. I have a Yeast Forge from Digital Homebrew a could not be more pleased with it. I have a 3400ml starter in a 5000ml flask going as we speak.

 

[butterpants]

You could build a hot box with some cardboard, tinfoil n heating pad....

 

[jwalkermed]

Question for you experienced people.

When I step up a colony or slant to make tubes to store in the freezer I have been using wort made from DME sterilized in a pressure canner. I get quite a bit of hot break and when I cold crash the yeast, add glycerol mixture etc... i end up with a good bit of hot break in the centrifuge tubes. I know this probably isn't a huge deal but ...

Do any of you just boil your step up media similar to a starter rather than sterilize it? Is this bad practice for yeast banking? I'd definitely get much less hot break material that way.

 

[bionut]

You should boil the wort on the stove first, and separate from the trub, then pressure cook. You can go with only boiling, but you never know what can resist the boil.

 

[mattrox]

I just made my 1st deposit in the yeast bank.

View attachment 1476442592683.jpg

 

[butterpants]

I autoclave my wort mixture in reagent bottles 50% full then once cool (and needing sterile wort), I manually pipette out what volumes needed....being careful and keeping the stock bottle sterile. I've had a stock bottle of wort for 2 months before. There's a significant amount of break material on the bottom but I just let gravity settle it and don't transfer it.

In a nutshell, I'd try to limit break material in your master culture vials. It looks ****ty and could possibly alter the freezing dynamics? But mostly cuz it looks ****ty.

I also wouldn't trust a normal boil alone for this kind of sterilization work.

 

[plowboy25127]

Just thought I'd show what I picked up at work today to keep my yeast in. I might have added an alarm to it just cuz we had one laying around....View attachment ImageUploadedByHome Brew1478539033.438799.jpg