Author Topic: Shaken not Stirred  (Read 7814 times)

S. cerevisiae

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Re: Shaken not Stirred
« Reply #30 on: February 11, 2015, 03:08:12 AM »
That was an interesting read.  Did you see this part at the end?

"Yeast cells in the exponential phase had weak turgor pressure compared to turgor pressure of stationary phase yeast cells."

"Both groups postulated that the ability of a yeast strain to support a higher gradient of osmotic pressure could be due to turgor pressure and thickness of the cell membrane. Cells, which normally have high turgor, have membranes with higher tensile strength. Thus, stationary phase cells, which have been reported to be more stress tolerant than actively growing exponential phase cells, must have a more rigid cell membrane in order to maintain the same volume than exponential cells."

I have mentioned that yeast cells undergo morphological changes at the end of fermentation in preparation for hard times in several thread on this forum and others. 

https://www.homebrewersassociation.org/forum/index.php?topic=21705.msg275241#msg275241

"By stepping a culture at the end of the exponential phase, we are pitching yeast cells that require very little in the way of replenishment. Hence, we will experience a shorter lag phase than we will if we pitch a culture that has reached quiescence because a quiescent culture has to undo the morphological changes it underwent in preparation for hard times.  A quiescent culture also has to replenish the ergosterol and UFA reserves that were spent post-exponential phase, which increases dissolved O2 requirements."

https://www.homebrewersassociation.org/forum/index.php?topic=21952.msg279097#msg279097

"As I have mentioned many times, a high krausen pitch will always outperform a "fermented out" pitch because the cells are in peak health and do not have to undo the morphological changes that occur at the end of fermentation.  Add in sufficient aeration and a strain with a high attenuation rate, and an AA of 85% is not out of the question."

I have also mentioned that cell wall thickening is the major morphological change.

https://www.homebrewersassociation.org/forum/index.php?topic=20692.msg262969#msg262969

"At the end of fermentation, yeast cells go into survival mode where their cell walls thicken and they store carbohydrate as glycogen.  In effect, the cells are preparing for hard times.  It takes longer to exit this state than its does when the yeast cells are still in active growth mode; hence, lag times in addition to oxygen demands are also increased."

What the authors did not mention is that this survival mechanism has to be undone during the lag phase; otherwise, nutrients and waste products cannot easily pass through the cell wall and plasma membrane.  Hence, stationary phase cells are no more likely to survive the lag phase than yeast cells that are pitched during the exponential phase, which is why we pitch at a higher rate.

Offline Philbrew

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Re: Shaken not Stirred
« Reply #31 on: February 11, 2015, 03:30:18 AM »
Sorry, spell checker erased an L in my post.  I wanted to say that I want to make two 2L starters (to pitch in two six gal. lager batches).  I don't have four 1 gal. jugs.

Headspace is critical to achieving a proper shake.  One is attempting to turn the wort into as much foam as is humanly possible.  This method is designed to be low-tech and low-cost. 

With that said, another thing that I am attempting to dispel is the insane notion that people have to hit the cell counts provided by yeast calculators in order to have a healthy fermentation.   It is better to have 60 billion healthy, ready to go to war with the wort cells than it is to have 200 billion stressed cells that are barely clinging to life.  The difference between 60 billion cells and 200 billion cells is approximately 180 minutes of propagation time.
OK, light bulbs are turning on all over.  "as much foam as is humanly possible"  makes big time sense because foam has huge surface area. 
This engineer has always believed that "low-tech and low-cost" are the better solution. 
I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.
Thanks, Mark.  Very enlightening post on how yeast do their thing.
Many of us would be on a strict liquid diet if it weren't for pretzels.

Offline narcout

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Re: Shaken not Stirred
« Reply #32 on: February 11, 2015, 04:18:05 AM »
What I hadn't heard before was the part about stationary cells being reported to be more stress tolerant.

I haven't had a chance to read the article cited at the end of that sentence, but here's the link.  I have no idea if it's interesting or presents anything that you haven't already mentioned.  My guess is that you've probably read it before.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC372915/pdf/microrev00025-0099.pdf

I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.

For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.
There's nothing left to dismantle; the house it just collapsed on itself.  - A. Savage

S. cerevisiae

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Re: Shaken not Stirred
« Reply #33 on: February 11, 2015, 05:44:57 AM »
What I hadn't heard before was the part about stationary cells being reported to be more stress tolerant.

The morphological changes occur in order to harden the cells against the harsh reality of living in a toxic, low-nutrient medium for an unknown amount of time.
 
Quote
For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.


Cell count is only half of the equation.  Cell health is equally, if not more important.  When pitching cells at high krausen, one is pitching very healthy cells with very pliable cell walls that require little to no maintenance before they can go to work; therefore, reducing lag time.  The number one bogeyman with normal gravity fermentation is house infection.  Shortening the lag phase shortens the time that bacteria have to multiply (as does reducing the number of replication periods), and bacteria multiply three times faster than yeast.  These growth rates are what we are up against in a real world brewery.

https://www.homebrewersassociation.org/forum/index.php?topic=19850.msg277460#msg277460

"A small amount of bacteria can overtake a much larger amount of yeast because the bacteria cell population increases 8-fold every time the yeast cell population doubles.  If we were to normalize the propagation period between yeast and bacteria (bacteria multiplies three times faster than yeast), the growth equations would be:

yeast_cell_count = initial_cell_count * 2n, where n = elapsed time in minutes since the end of the lag phase / 90

bacteria_cell_count = initial_cell_count * 8n, where n = elapsed time in minutes since the end of the lag phase / 90


If we run the numbers, it should become crystal clear why one wants to pitch a large, healthy yeast culture while doing everything possible to minimize the opportunity for bacteria to catch a ride into one's yeast crop, starter, or fermentation vessel.  It should also become clear why the growth phase is called the exponential phase.


Cell counts at 90 minutes

yeast_cell_count = initial_yeast_cell_count * 21 =  initial_cell_count * 2
bacteria_cell_count = initial_bacteria_cell_count * 81 = initial_cell_count * 8


Cell counts at 180 minutes

yeast_cell_count = initial_yeast_cell_count * 22 =  initial_cell_count * 4
bacteria_cell_count = initial_bacteria_cell_count * 82 = initial_cell_count * 64


Cell counts at 270 minutes

yeast_cell_count = initial_yeast_cell_count * 23 =  initial_cell_count * 8
bacteria_cell_count = initial_bacteria_cell_count * 83 = initial_cell_count * 512


Cell counts at 360 minutes

yeast_cell_count = initial_yeast_cell_count * 24 =  initial_cell_count * 16
bacteria_cell_count = initial_bacteria_cell_count * 84 = initial_cell_count * 4096


Cell counts at 450 minutes

yeast_cell_count = initial_yeast_cell_count * 25 =  initial_cell_count * 32
bacteria_cell_count = initial_bacteria_cell_count * 85 = initial_cell_count * 32768


Cell counts at 540 minutes

yeast_cell_count = initial_yeast_cell_count * 26 =  initial_cell_count * 64
bacteria_cell_count = initial_bacteria_cell_count * 86 = initial_cell_count * 262,144


Cell counts at 630 minutes

yeast_cell_count = initial_yeast_cell_count * 27 =  initial_cell_count * 128
bacteria_cell_count = initial_bacteria_cell_count * 87 = initial_cell_count * 2,097,152


Cell counts at 720 minutes

yeast_cell_count = initial_yeast_cell_count * 28 =  initial_cell_count * 256
bacteria_cell_count = initial_bacteria_cell_count * 88 = initial_cell_count * 16,777,216"



S. cerevisiae

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Re: Shaken not Stirred
« Reply #34 on: February 11, 2015, 05:47:32 AM »
OK, light bulbs are turning on all over.  "as much foam as is humanly possible"  makes big time sense because foam has huge surface area. 

Exactly!

Offline Philbrew

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Re: Shaken not Stirred
« Reply #35 on: February 11, 2015, 07:43:09 PM »

I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.

For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.
[/quote]
Sorry, I didn't mean to infer that the Mr. Malty calculator is inaccurate.  In fact it's probably the best.  It's just that all the yeast calculators seem to be overly cautious on yeast production rates except for stir plates. I realize that it would be difficult to add to the calculator the Shake-it-until-it's-all-foam method or combine O2 at start plus shaking.   But, as it is, the calculators tend to push one into buying a stir plate, especially if you want to do lagers or big beers.  I want to brew lagers and the $200 for a heavy duty stir plate and 5L flask could be better spent on a wort chiller or a temp controller for my fridge and an upgrade of my brew pot.
Many of us would be on a strict liquid diet if it weren't for pretzels.

Offline TMX

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Re: Shaken not Stirred
« Reply #36 on: February 11, 2015, 07:48:25 PM »

I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.

For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.
Sorry, I didn't mean to infer that the Mr. Malty calculator is inaccurate.  In fact it's probably the best.  It's just that all the yeast calculators seem to be overly cautious on yeast production rates except for stir plates. I realize that it would be difficult to add to the calculator the Shake-it-until-it's-all-foam method or combine O2 at start plus shaking.   But, as it is, the calculators tend to push one into buying a stir plate, especially if you want to do lagers or big beers.  I want to brew lagers and the $200 for a heavy duty stir plate and 5L flask could be better spent on a wort chiller or a temp controller for my fridge and an upgrade of my brew pot.
[/quote]

on uline.com, you can get a 4 pack of 1 gallon glass jugs for less than $20 (plus shipping), split a single vile 4 ways, and have a 1 gallon starter.....no stirplate needed.

T
"The ART of brewing Beer, is the ACT of brewing Beer"
https://txbrewing.wordpress.com

Ferm 1: Irish Red Ale
Ferm 2:

On Deck: American Wheat

Keg 1: Un-Common
Keg 2: Switchback Stout

Total Gallons brewed (2015) - 10

S. cerevisiae

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Re: Shaken not Stirred
« Reply #37 on: February 11, 2015, 08:16:33 PM »
on uline.com, you can get a 4 pack of 1 gallon glass jugs for less than $20 (plus shipping), split a single vile 4 ways, and have a 1 gallon starter.....no stirplate needed.

Just remember to shake the jugs like they owe you money (think mafia enforcer)! :)

Offline narcout

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Re: Shaken not Stirred
« Reply #38 on: February 11, 2015, 08:20:27 PM »
It's just that all the yeast calculators seem to be overly cautious on yeast production rates except for stir plates.

Yes, that could very well be true (or perhaps they are overly optimistic on the effects of stir plates). 

I believe at least some of the predicted results from the Mr. Malty calculator are based on real world observations, but I do not know to what extent (though this info may be out there on the internet somewhere).

Fortunately, as has been pointed out, you do not need to be exact in your pitching rates.  They are just another tool that you can use to achieve your intended flavor profile.

By the way, another good source for inexpensive one gallon glass jugs is your local supermarket (and they come filled with delicious juice).

There's nothing left to dismantle; the house it just collapsed on itself.  - A. Savage

Offline Philbrew

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Re: Shaken not Stirred
« Reply #39 on: February 11, 2015, 08:44:38 PM »
on uline.com, you can get a 4 pack of 1 gallon glass jugs for less than $20 (plus shipping), split a single vile 4 ways, and have a 1 gallon starter.....no stirplate needed.

Just remember to shake the jugs like they owe you money (think mafia enforcer)! :)
Yup, that's my plan.  Hmmm...I wonder if my local Ace Hardware would let me put them on their paint shaker? ;)
Many of us would be on a strict liquid diet if it weren't for pretzels.

Offline TMX

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Re: Shaken not Stirred
« Reply #40 on: February 11, 2015, 09:39:48 PM »
I need to take a photo what the amount of yeast that I pitch into 600 milliliters of wort (my primary volume is 3.5 gallons), but it is a ridiculously small amount of yeast compared to what is in a White Labs vial.  You will laugh at the thought of ever thinking that half of a White Labs vial is not enough yeast to pitch into a 1L starter when you see how little yeast I pitch.

With that said, if my math is not failing me, 2 tablespoons is just shy of 30ml. Thirty milliliters of truly thick slurry can have up to 3 billion yeast cells per milliliter, or up 90 billion yeast cells in total.  I kid you not when I say that you can pitch five gallons of ale with 90 billion fresh viable cells with no off-flavors or loss of attenuation as long as the wort is properly aerated.

you keep threating us with this....lol

"The ART of brewing Beer, is the ACT of brewing Beer"
https://txbrewing.wordpress.com

Ferm 1: Irish Red Ale
Ferm 2:

On Deck: American Wheat

Keg 1: Un-Common
Keg 2: Switchback Stout

Total Gallons brewed (2015) - 10

S. cerevisiae

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Re: Shaken not Stirred
« Reply #41 on: February 11, 2015, 10:07:22 PM »
you keep threating us with this....lol

I inoculate one of these media bottles with one or more 4mm loop scrapes taken from a slant. 



The yeast that is grown in 40 milliliters of absolutely sterile (autoclaved) wort is pitched into 600 milliliters of 10% w/v (1.040) wort.  That's a one to fifteen step media-wise, which is living on the edge when dealing with such a small amount of yeast.   A White Labs vial contains almost 35ml of yeast cells.  I am pitching at best 1/10th that amount.

Offline TMX

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Re: Shaken not Stirred
« Reply #42 on: February 11, 2015, 10:22:47 PM »
i need to see if they have autoclaves on Craigslist
"The ART of brewing Beer, is the ACT of brewing Beer"
https://txbrewing.wordpress.com

Ferm 1: Irish Red Ale
Ferm 2:

On Deck: American Wheat

Keg 1: Un-Common
Keg 2: Switchback Stout

Total Gallons brewed (2015) - 10

S. cerevisiae

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Re: Shaken not Stirred
« Reply #43 on: February 12, 2015, 01:29:49 AM »
i need to see if they have autoclaves on Craigslist

You do not need to purchase a laboratory autoclave.  A pressure cooker/canner that is capable of 15 PSI above normal atmospheric pressure will get the job done.  You need to be careful when purchasing because many modern pressure cookers/canners are not capable of achieving 15 PSI above normal atmospheric pressure pressure.  All of the "All American" pressure cookers/canners are capable of achieving this pressure level.  A few of the lower-priced pressure cookers are capable of achieving 15 PSI above normal atmospheric pressure as well.

Offline TMX

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Re: Shaken not Stirred
« Reply #44 on: February 12, 2015, 04:49:18 AM »
This has been sitting at 66 since Sunday night, I have to leave town Fri-Mon, would there be any benefit it raising the temp up to 70ish while I am out of town
"The ART of brewing Beer, is the ACT of brewing Beer"
https://txbrewing.wordpress.com

Ferm 1: Irish Red Ale
Ferm 2:

On Deck: American Wheat

Keg 1: Un-Common
Keg 2: Switchback Stout

Total Gallons brewed (2015) - 10