Is it possible to use a yeast starter too early?

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Johnwongfat

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I fired out a couple of yeast starters late last night and I wanted to use one today for brewing. They've been sitting for about 8 hours now and there is a small layer of foam on the top (maybe 1/4"). My other 2 starters had sat for about 2 days and had about 1-2" of foam residue on the sides. My question is if I use it today am I going to kill the yeast? Is this a complete waste of a starter, patience is not a virtue I possess and I want to brew today, but I don't want to kill my brew. Any thoughts?
 
There are many reason for making a starter, but one is to verify the yeast is good. If you have activity already then you know it's good.

Go ahead and pitch it ("pitch" - to add yeast to the wort, not "pitch" to throw out). :D
 
The short answer: no. Eight hours isn't much time for you or I, but in a good starter that's three or four doublings. So, you have 8-16 times as much yeast as you started with.
 
I won't disagree with you that it may not be the desired population of yeast, but the question was "is it too early". Under an "optimum" situation the answer would certainly be "no".

But at this point he has a viable yeast and he wants to brew now. I still stand by my answer to his question - "no" it is not too early.

Overall, he is in a much better position to pitch his yeast than someone who just pitched dry yeast of did not make a starter. :D

He'll just have to plan better next time, or put off brewing until tomorrow :( which may not be possible for him.

It's OK to have different brewing techniques. We all pick and choose which techniques are best for us (grain vs DME vs kit) and we all learn where we can take a shortcut here or there (2 oz of 5%AA vs 1 oz 10%AA), dry yeast vs a full blown liquid yeast starter, but in the end it's us brewing something we like whether or not we want to imitate a style of modifiy it to our tastes. :D

Happy Brewing!
 
I've pitched WL straight from the bottle & had very good results. But, more yeast is better (yah, I know adding a gallon of yeast to a 5 gallon batch is a bad thing, but who ever does that?)
 
Johnwongfat said:
So conversly, is it possible to use my starter too late? I.e let it sit out all week. Will this kill my yeast or does it just increase the likelihood of contamination?

I wouldn't recommend it unless you are planning to keep feeding it to increase the yeast cell yield. You can add more wort daily to keep it producing.

Alternatively, you could grow your starter and then refrigerate it for a week, warm back up and add more wort to restart it.

Or you could do as you said, I don't guess it's any different than a mini-primary a week shouldn't hurt a thing I just don't see a reason to do it that way.
 
Johnwongfat said:
So conversly, is it possible to use my starter too late? I.e let it sit out all week. Will this kill my yeast or does it just increase the likelihood of contamination?


This is what can happen:

Autolysis is the process by which a cell self-destructs for the healthiness of the entire organism. Autolysis occurs when a lysosome allows the digestive enzymes out of its membranes. The cell then, in effect, digests itself.


Not good.
 
So the moral of the story is, "if you make two starters be sure you're going to be able to use them both in the next 2-3 days." I just added more food to the second one, I used the other on Monday. I figure the extra yeast shouldn't hurt an imperial stout so I think I'll be ok. I guess I'll find out by tomorrow if the yeast is still viable. Thanks for all the info, still learning.
 
ORRELSE said:
This is what can happen:

Autolysis is the process by which a cell self-destructs for the healthiness of the entire organism. Autolysis occurs when a lysosome allows the digestive enzymes out of its membranes. The cell then, in effect, digests itself.


Not good.


That can't happen in one week though. Can it? I mean we let our primaries sit on the yeast and trub for a week. I thought that the process of autolysis happens after about 3 weeks.
 
I always make mine 24 hrs before I expect to pitch and it seems to work out fine. I can't see an advantage to letting it sit out for a week; if the scenario is you were expecting to brew and you didn't get the opportunity, then I would follow ScottT's procedure of refrigeration, although I think restarting it is optional...you could just decant the liquid and pitch the slurry. Depends on your religion.
 
I wouldn't worry too much about autolysis withing a weeks time. IIRC, Palmer recommends 3+ days for a liquid yeast starter to build up. Something about building up cell walls and reserves and stuff. I've never waited more then a day and have had decent results so far, so it would seem they're pretty forgiving.
 
Yes you could in theory use it too early (i.e. <2-4 hours after starting). However, what you are describing is fine probably early exponential growth!

Remember that there are basically four stages of yeast growth kinetics.

1. Lag phase. The time it takes the yeast to start going from transfer to a growth medium.

2. Log phase. This is where the yeast are growing at optimal growth rate and with good aereation/rich complete medium are the the best off. The yeast are undergoing exponential growth...the only time during the growth phase. During the log phase....twoard the end is best (for yeast #s) is the best time to pitch for optimal "steping-up". During this time the yeast are laying down "stores" (i.e. glycogen and others) for the impending next stage. Think early to full krausen.

3. Stationary phase. This is where the yeast slow down and basically stop growing they are still growing but slowly. Majority of the energy sources in the medium are used. During this time the yeast change their expression profiles and look to use other metabolic pathways to maintain their growth (if not put in a fridge). Stores such as glycogen are starting to be used. The yeast start to fall out of solutin and starts to floculate. Most of the time we pitch at early time points of stationary phase. As the full krausen starts to drop off is late log early stationary phase. The length of the stationary phase is varible and depends on the stores/yeast health from the growth phase and the growth medium.

If you "force" a stationary phase...probably "slow growth" phase would be the best term with excess sugar the yeast will remain viable from 2wks to 1 months in the fridge. Heck 3 to 6 months is doable but viability suffers greatly. Of course using earlier is better. All you need to do is warm up to room temp for 8-12 hours. The yeast will start going again....make sure to aereate when cold to increase [O2] in the wort.

4. Death phase. The yeast start to die/autolysis occurs. The yeast population has used all of their nutrient sources and die. The release of the stores can be used for a short time by other yeast....almost like cannablism but if nothing happens the colony will ultimately die. Can happen quickly or take a long time....depend on the yeast.

For a starters (1.030-1.045) pitched with healthy yeast the stages are fairly rapid at room temp (from what I've seen).....lag is about 1-2 hours; log is 2-24 hours; stationary starts around 24-48 hours post pitch and can last about 2 days; death phase starts between 3-5 days post pitch. Of course this depends on the medium, yeast health, and numbers of viable yeast pitched.

-Eric
 
That Death phase troubles me a little. 3 to 5 days you say? What prevents the autolysis from happening in the bottle when bottle conditioning? Likewise, how do White Labs and Wyeast prevent their yeast from entering the death phase in their 3 month shelf life?
 
ScottT said:
That Death phase troubles me a little. 3 to 5 days you say? What prevents the autolysis from happening in the bottle when bottle conditioning? Likewise, how do White Labs and Wyeast prevent their yeast from entering the death phase in their 3 month shelf life?


Amount of yeast affects autolysis. Death phase does not equal autolysis but an increase probablity of autolysis. Autolysis can occur at all stages but is associated more with the death phase. The rate of autolysis is not fixed and increases with time is also associated with highly flocculant yeast (due to a range of factors associated with mean colony size). Amount of yeast in the associate percipitate also affect autolysis. There are relativly few yeast in the bottom of the bottle (due to the secondary) and they are stored under pressure usually cold. Not optimal conditions for autolysis....but it can still occur.

Where does White labs and wyeast store their yeast? Its in the fridge. Cold affect yeast metabolism and allows for long term storage. The longest I've been able to store a yeast in the fridge is 2 years (sealed of course).

-Eric
 
LOL, this hobby is becoming increasingly more complex. :) Everytime I think I'm getting my head around it, someone comes along and shows me I don't know jack. I re-started my yeast last night and got about 1/4" of foam today, which has mostly subsided by now (about 24 hours since their feeding). I had one starter that actually overflowed my container so I am a bit worried that I may have killed my yeast now. I've read some posts that say 1/4 - 1/2 inch is normal krausen for a starter, is this true? I was hoping to use the yeast tomorrow but I don't want to ruin my first imperial stout. Is it safe to pitch my yeast or should I just cough up the 5 bucks for more yeast?
 
Johnwongfat said:
LOL, this hobby is becoming increasingly more complex. :) Everytime I think I'm getting my head around it, someone comes along and shows me I don't know jack. I re-started my yeast last night and got about 1/4" of foam today, which has mostly subsided by now (about 24 hours since their feeding). I had one starter that actually overflowed my container so I am a bit worried that I may have killed my yeast now. I've read some posts that say 1/4 - 1/2 inch is normal krausen for a starter, is this true? I was hoping to use the yeast tomorrow but I don't want to ruin my first imperial stout. Is it safe to pitch my yeast or should I just cough up the 5 bucks for more yeast?


Don't worry about an overflow, just keep it covered the best you can to avoid contamination. The remaining yeast are just fine.
 
eolle said:
Amount of yeast affects autolysis. Death phase does not equal autolysis but an increase probablity of autolysis. Autolysis can occur at all stages but is associated more with the death phase. The rate of autolysis is not fixed and increases with time is also associated with highly flocculant yeast (due to a range of factors associated with mean colony size). Amount of yeast in the associate percipitate also affect autolysis. There are relativly few yeast in the bottom of the bottle (due to the secondary) and they are stored under pressure usually cold. Not optimal conditions for autolysis....but it can still occur.

Where does White labs and wyeast store their yeast? Its in the fridge. Cold affect yeast metabolism and allows for long term storage. The longest I've been able to store a yeast in the fridge is 2 years (sealed of course).

-Eric

My questions about pitchable yeast and bottle conditioning were retorical questions. My point is was that autolysis does not happen in 3 to 5 days just because yeast enters the death phase.

What troubled me was the fact that you and Orrlese seem to be claiming that a yeast starter can autoloyse in 1 week. I find that highly unlikely given that we are able to bottle condition on sedement for months and months, unrefigerated without autolysis.

In this most recent explination, you say that the amount of yeast in the precipitate, the degree of flocculation, the amount of pressure and the temp all play a role in autolysis.

So given all these variables, is there a group of charts somewhere that can predict the the risk of autolysis considering the cell count, degree of flocculation, the pressure and the temp in relationship to time?

A 5 gal. carboy with over 10 million cells of yeast per cl finishes fermenting, flocculates and settels in 3 days, You leave it for a week and it's been in the death phase for over 3 days. Most of us pull it off to a secondary at that time but many do not. Are their yeast autolyzing at 70 degrees F under 1 atmosphere of pressure, highly focculated, and in the death phase for 10 days?

Do you understand why I question this? What your are trying to teach me just doesn't seem to match up with what I see taking place. At least it's clear as mud to me.
 
Growth phases is different than autolysis. I presented no contridictary material and I never stated that autolysis was a big deal...just that it occurs. I think this is where homebrewers are just paranoid about a term that will rarely affect the beer. Don't fear the yeast reaper :)

Autolysis occurs at all stages...the percent and amount autolysis is what is important. 1 cell in 3x10^6 cells per day will not be perciptable for a long time lets say....5 years. 10 -100 cells in the same yeast cake would take 1 year to be percieved. It occurs. Cell numbers decrease...they have to go somewhere. Autolysis in small numbers is not that big of a deal and the majority of the released contents will be absorbed by other yeast.

Just because autolysis occurs it does not mean it will be perceptible. In fact autolysis is a mechanism of flavoring in some baked goods. Where autolysis is a big deal....is if there is a sudden event where a large numbers of cells enter at the same time. It can actually form a "run-away" type event where there are enough lysosomal enzymes to weaken/destroy other cells and continue the cycle. This is very very rare in a homebrew situation and requires a major event (i.e heat, detergent, sometime bacterial contamination ect).

One think I find interesting...and needs to be studied....is the role of the fementer shape on autolysis. I think a 5-6 gal carboy, bottle anything with a wide flat bottom is not as prone to major autolysis events. This is probably due to nutrient diffusion? However, fermenters that seem to increase yeast bed/trub thickness (i.e. conical) could cause an increase probablity of major autolysis....that needs to be studied.

Good link on yeast starter. Mostly designed for industrial apps. since we homebrewers don't really need to add glycerol, peptone or other stuff to get maximal density and most don't have access to shakers. It a lot like the lab handout I wrote for my students. I only added trypan blue dye exclusion to teach/measure viability. I'll have to see if I still have the handout...it was about 5 years and a couple of computers ago :)

-Eric

autolysis...autolysis...autolysis...autolysis...autolysis...see it didn't hurt anything :D Its just a word and a standard process that can only hurt if you are affraid of it. :D
 
There's something about the death phase that's actually useful to the culturing of yeast. I'm going to have to dig it up from somewhere though. The gist of it was that during that time the yeast cells go into a sort of protective state, in which they are actually better prepared to enter their new environment (ie your wort).

Eric, do you know off-hand if its during this death phase that the yeast become less specialized, in the sense that the production of enzymes they've been producing for their particular environment clear up, essentially giving them a "clean slate" of sorts? I'll see if I can find my reference.
 
Okay, this is what I was referring to:
John Palmer said:
[...] towards the end of fermentation, yeast build up their glycogen and trehalose reserves; kind of like a bear storing fat for the winter. Glycogen and trehalose are two carbohydrates that act as food reserves for the yeast cell. Yeast slowly feed off these reserves when other food is not present, and use this food extensively to fuel the synthesis of essential lipids, sterols, and unsaturated fatty acids when pitched to an oxygenated wort. (Yeast will rapidly deplete their glycogen reserves when exposed to oxygen.) While glycogen can be likened to the fat that a bear stores for winter, the other component, trehalose, acts more like the bear's heavy fur coat. Trehalose seems to get built up on both the inside and outside of the cell membrane, and is generally believed to make the membrane structure more robust and more resistant to environmental stresses. By allowing the yeast starter fermentation to go to completion, these reserves are built up, and upon pitching, the yeast starts out with a ready fuel supply and a clean slate to better adapt it to the new wort. As noted earlier, though, these same reserves are used by the yeast while in hibernation, so if the yeast are left too long before pitching, the reserves may be depleted and should be replenished with a fresh starter wort fermentation before use.
I'm a little confused on exactly when this takes place though. Is this occuring during this "death phase", or earlier?
 
Glycogen in healthy cells is made during the log growth phase into early stationary phase. During stationary and death phase the glycogen store...a good form of energy is used by the yeast. It is the "fat" stores.

Trehalose is not a very good source of metabloic energy. It can be used but there are better sources of rapid energy. It is made straight through the stages....but most importantly during thermal shock and later stages. Trehalose actually is a really cool sugar. It is believed to act to stabilize protein structures and act as a free radical scavenger. Here is some work from a really good Japanese group:

http://aem.asm.org/cgi/content/full/64/11/4614

and some work showing the stabilization of yeast in ethanol by trehalose

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8172916&dopt=Abstract

I have used it for this purpose before and a lot of companies use trehalose to stabilize purified proteins. I'll leave you with one quote on trehalose from Goldberg:

"Many students of yeast metabolism in the past had studied trehalose and assumed that it serves as a possible energy reserve," Goldberg said, "but its major role in our work is as the natural cryoprotectant that appears to protect proteins against unfolding and against oxygen radicals during freezing and at high temperatures."

Yeast can undergo a dormant state. Usually associated with changes in the cell wall and increased trehalose and other cellular products. We can easily induce in the cold. Also while it is rare in beer (I guess it could be possible), yeast can form acrospores....a highly tolerant form but then we start to get to meiosis and yeast sex (no kidding...yeast have "sex").

HTH

-Eric
 
Oh now I understand. I was under the impresson that if your yeast autolized, the flavors this caused would render your beer unsuitable for consumption. Glad to hear otherwise.
 
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