What to Post after a Long Hiatus?

An Experiment (Yeast
Aeration) of Course!
My first post was in 2010, a standard bitter in honor of close colleague and astounding scientist that passed. I was in my fourth year
as a post-doc at Columbia University studying how retroviruses (HIV, MLV) interact with the cells they try to infect. Time was in
abundance, allowing me to brew in a small Bronx apartment while I gathered many friends and scoured the city for rare-to-find craft
beers. I was able to ranch yeast and create a bank, isolate uniqueBrettanomyces strains, and collaborate with a world renown master
brewer. Most importantly, I learned as much as I could about the science behind brewing beer. My time in academic research came
to an end, and I married the most amazing woman I have ever known and met another one:
Natalie, my beautiful daughter, was a game changer for me. I assume that most fathers sense of their place in the world changes
dramatically when your first-born looks straight into your eyes. Job hunting was next, and I was lucky enough to find a great job
doing process development for vaccines at Merck. Moving and settling in, enjoying fatherhood, and trying to secure my familys
future has made me feel slightly old.
As a result of all of this stuff, my last post was back in May. I began to re-read some of my old posts and realized that this blog has
been is linked to so many wonderful memories, both beer and non-beer events that I cant possibly ignore it anymore. Its posts
serves as markers for my past. It is this for this reason I intend to start posting again.

And an experiment may be the best way to start posting again. As a recap, I began brewing experiment beers for a yeast class that I
taught at Brooklyn Homebrew. Ben and Danielle were kind enough to provide resources and space for my scientific curiosities that
kept brewing. In contrast to my work life, my style of experiments are not rigorous and grounded in statistical significance. The goal
has always been to pique the interest of the vast majority of non-scientific brewers, both professional and amateur. More
importantly, I do not have the resources at home to measure key components of the brewing process. Instead, my conclusions rely
on the most important data point the beer drinker.
I am teaching again, this time at a great homebrew store in PA Keystone Homebrew. Jason Harris, along with Lou (store manager)
has been kind enough to provide support and space for the yeast class and the experiment beers. Ive already taught two classes, and
the most recent one was this past Saturday (November 9th).
Oxygen
Yeast are facultative anaerobes, meaning that they prefer to use oxygen to produce energy efficiently through oxidative
phosphorylation. In the absence of oxygen, yeast still produce energy but with the inefficient process of alcoholic fermentation.
Luckily for brewers, yeast will almost always undergo fermentation due to the Crabtree effect, where in the presence of excess
sugars, even if there is oxygen, yeast will produce ethanol and CO2.
The main reason yeast need oxygen is for the production of unsaturated fatty acids (UFA) and sterols. Both molecules make up the
plasma membrane of the yeast cell. I wont go into how they are synthesized, but sterols, such as ergosterol pictured below, is
present in the cell membrane at much lower concentrations. Sterols provides fluidity, making the membrane easier to bend and
twist in shape.

Without sterols, UFAs would form rigid micelles, or droplets of fats. Importantly, this membrane fluidity allows for other vital
proteins to be imbedded in the membrane. Some of these proteins are responsible for importing not only complex sugars (maltose,
maltotriose), but vitamins, minerals, and cofactors. With poor cell membranes yeast will not tolerate increasing amounts of ethanol

(which is why oxygenating that strong ale is crucial). Lastly, if the cell membrane is not healthy enough, bud scars from cell budding
will take a toll on yeast health and restrict future growth.

What are the

consequences of introducing too little or too much oxygen? It is best to think of it biochemically that is, oxygen acts as an
accelerant to certain biochemical pathways. For example, what I did not mention before is that O2 drives the synthesis of UFAs and
sterols through a cofactor called Acetyl CoA.

I talk about this molecule a bit in my post on esters and it sits at the crux of critical yeast biochemistry. In this case, oxygen drives
acetyl CoA to form lipids (UFAs, sterols). As more acetyl CoA is used to make UFAs, less is used to make esters and fusel alcohols.
Therefore, beers that have more oxygen tend are cleaner in esters and fusels (think IPAs). Beers that have less oxygen will have more
esters and fusels and are more fruity (think English ales). Increasing oxygen will grow more yeast cells as their membranes are super
healthy and ready for budding. The increase in yeast biomass will add a bump in attenuation and may even thin out the beer since
the yeast will be so active.
How much Oxygen?
Homebrewers have endless methods of adding oxygen such as shaking and splashing the wort, sloshing the wort from bucket to
bucket, aeration with aquarium pump, and adding pure oxygen. I have heard lots questions since many homebrewers practice
different techniques:

How long do I shake?

Should I use a drill with a whirl attachment?

How long do you aerate with a pump?

How long do you pump in pure O2? At what flow rate?

Let me answer all of these questions by saying that whatever works for your system that gives you the best beer is what you should
do. Experimenting here is key. Brew that same pale ale three times, one with splashing, one with pure O2 (one minute), and one
with pure O2 (five minutes).
In reality, as homebrewers it is very difficult not to introduce O2 once the wort is cooled. Simply transferring the cold wort into a
carboy (without splashing) will add some amount of oxygen for the yeast to use, however this is far from ideal. Yeast need greater
than 8 parts per million (ppm) to adequately ferment a batch of beer. Here are some numbers on different methods (keep in mind
that O2 dissolves less in higher gravity wort) on an average batch of homebrew (5 gallons, 1.050 1.070):

Shaking: 2 ppm

Aquarium pump: maximum of 8 ppm

30 seconds pure O2: 5 ppm

60 seconds pure O2: 8-10 ppm

2 minutes pure O2: greater than 14 ppm

The above pure O2 is assumed to be one liter per minute.

Splashing the wort is a very ineffective method of introducing oxygen and may even introduce contamination. Notice that with an
aquarium pump, air can only give you a max of 8 ppm, whether you pump for five or thirty minutes. Oxygen makes up only 21% of
dry air so there is only so much O2 you can force into beer with this method. Moreover, 30 minutes of aquarium pumping may
drastically reduce head formation since you will create so much foam. The only way to get enough oxygen into your wort is by using
pure oxygen with a sintered stone.
Exactly how much O2 to add to your beer as this depends on many factors:

Yeast strain (different O2 requirements for different strains)

Wort gravity (need more O2 for higher gravity worts)

Beer style

Beer style and flavor profiles generated from oxygenation is the most important factor to me and is the basis of my experiment. For
example, will adding less oxygen enhance fruity characters of some english ales? Would my IPAs be cleaner with higher amounts of
O2? Would there be any changes in fusel alcohols? Would the body be thinner or fuller? Textbooks and the internet can tell you the
answers, but I feel it is better to experiment and find out for yourself.

For the experiment beer, I brewed an ESB in a ten gallon batch at Keystone Homebrew (OG: 1.056 with 40 IBUs):

1 lbs Crystal Light 45L (Thomas Fawcett)

1 lbs Crystal Malt 90L (Thomas Fawcett)

8.0 oz Pale Chocolate Malt (Thomas Fawcett)

12 lbs DME Golden Light (Briess)

2.00 oz Target [11.00 %] Boil 60.0 min

2.00 oz Goldings, East Kent [5.00 %] Boil 0.0

1.0 pkg British Ale II (Wyeast Labs #1335)

From this wort I split the batch four ways and provided different amounts of oxygen:
1.

30 seconds of splashing the carboy

2.

45 seconds O2

3.

5 minutes O2

4.

10 mgs of Olive oil

Sample (1) represents what a new homebrewer might do. I did not dump the wort between two buckets but rather sloshed the wort
around in the carboy. This sample should have little dissolved O2. Sample (2) is my default oxygenation regimen at home I usually
add pure O2 for 1-2 minutes in a 5 gallon batch. Sample (3) was to observe any changes in the other direction with large amounts of
oxygen added to the beer. When I did oxygenate this batch, the foam generated was huge, almost a 3 gallon tick mark. Unfortunately
I did not have access to a medical grade O2 regulator and was unable to determine flow rate. I opened my regulator, what you
typically find at homebrew stores, to its max setting.
Sample (4) may be the most interesting of all. I decided to mimic what Grady Hull, brewmaster at New Belgium, did for his masters
thesis at Heriot-Watt University. The thesis can be found here and is generously hosted bybrewcrazy.com. In a nutshell, he
added dissolved olive oil to the yeast starter of a commercial batch of Fat Tire as a source of linoleic acid, a precursor to UFA
synthesis and did not aerate the wort. The reasoning being that the oil provides all the UFA and sterol needs of the cell and you dont
need to add O2. The goal of the experiment was to prolong beer half-life since oxygen has a severe staling effect. What they found
was that the beer took a bit longer to ferment and was slightly more fruity. Independent tasters actually preferred this beer over the
control beer.
For my experiment I added olive oil directly into the wort (no starter were made for each sample). Importantly, the amount of olive
oil I added followed close to what Gary Hull did (1 mg per 25 billion yeast cells), which was exceedingly small (about a tenth the size
of a pinhead). Moreover, olive oil will not dissolve in wort and must first be dissolved in 100% ethanol. For any homebrewers
reading this do not add a drop of olive oil to your beer. I had to weigh 1 gram of oil, dissolve it, and make a serial dilution until I
had close to 0.1 ug/ml. Of this solution, I added 100 uls directly to the wort.
The beer is already fermented, bottled and was sampled at a yeast class this past Saturday (11/9/2013) at Keystone Homebrew.
Similar to my past experiments, I will post flavor profiles from the students as data points in a future post.

Pitching Rate Experiment Results

A few posts back I talked about yeast pitching rate, or the amount of yeast used to inoculate unfermented wort, and how critical this
number is to brewing great beer. Pitching not enough yeast or too much yeast can create off-flavors and make a subpar beer.
Moreover, the pitching rate itself is a number thats not set in stone but rather subject to different variables such as the strength of
the beer and the yeast strain selected. Brewers, both pro and amateur, change the pitching rate to suit their needs. A slightly higher
pitch rate for an ale may produce a completely different beer. The most important thing to do is experiment with your system and
determine the best pitch rate for a strain of yeast.
To this end, I did an experiment to find for myself the effects of over-pitching or under-pitching a beer on my brew system and the
results are in this post. This experiment created its own thread over at Homebrewtalk so I wanted everyone reading to get these
results. I want to point out, however, that I dont feel the experiment was a success, although some information was gleaned from the
experiment. Here are the reasons why it failed:
1.
2.

Beer style. I chose a blonde ale thinking that off-flavors would be more apparent. Differences were actually very subtle.
Yeast strain. This is probably the biggest mistake. I chose Wyeast 1056, or Chico yeast, to test. Unfortunately, this yeast is
pretty clean and pitching rate changes might not change the yeast flavor profile that much.

3.

No oxygen or aeration. I did not oxygenate the wort prior to pitching. I thought the action of sprayed wort hitting the side
of the fermentation vessel would introduce enough oxygen but I think I was wrong. Several brewers on homebrewtalk
pointed this out and I think they are right. The control beer had a strange off-flavor that was hard to place.

4.

Fermentation vessels were not covered. Another big mistake. There was varying degrees of lightstruck, with the control
pitch the most affected.

Having said all of this, here is the fermentation profile of the three beers:

I measured the gravity of each beer every 8 to 12 hours using a 200 ul pipette-man. As you can see, the fermentation profiles are
exactly what I was hoping for. The over-pitched beers rate of fermentation was faster than the control, while the under-pitched beer
was slower. One interesting thing to note was the under-pitch beer finished at a lower gravity than the control pitch (1.009 compared
to 1.012). Im not sure what this means and this bears repeating. Could under-pitching give slightly lower final gravities? One
speculation is that the yeast are stressed to the point where attenuation is raised.
Once the beers finished bottle conditioning, I had my wife, Kim, set up the samples in a blind tasting for me. I could not pick out the
control beer. Actually, I totally missed on guessing which beer was which. The next day, I took all the beers to a yeast class I was
teaching at Brooklyn Homebrew yeast and the students could not pick out the beers either. This was my first time teaching a class on
yeast for homebrewing, and I went over the 2 hour time limit. I was rushed, due to no fault but my own, and did not tally what
people thought of the beers.
However, after knowing which beers were which (I know not very controlled/blinded) I could taste subtle differences. Hopefully
my ability to critique my beer is not subject to suggestion. Here is my take on the three beers, faults and all:
Control-pitch: The nose smelled of faintly oxidized hops and medicinal. Uh oh. Toasty malt background with agressive bitterness
that approaches astringent. There is a phenolic, almost plastic-like taste in the backend. I dont think this is due to contamination as
the other beers seem fine. Its possible that the lack of oxygenated wort and light produced this result. However, cutting through this
reveals a very clean beer with the american two-row shining.
Over-pitch: No evidence of phenols. However, the is no evidence of anything. The beer is super clean with a bland malt profile. The
beer is thin and almost water-like with very little body.
Under-pitch: The nose of the beer gives away some fruiter ester that lingers and disappears. The taste has an odd assortment of
esters that clash with the malt and make the beer quite unpalatable. This beer had a thicker body (although slight) compared to the
control. The ester profile, although not strong for Wyeast 1056, reminded me over-ripe peaches.
Conclusions: As a scientist, I would have to say these results are pretty much inconclusive since my control did not work. However,
the over-pitch beer was cleaner and thinner while the under-pitched beer had some strange esters going on. This would seem to back
what other people have seen about pitching rates: over-pitching produces a bland beer, while under-pitching produces a stressed
beer with esters being produced from the yeast.
Future: I plan on teaching another class at Brooklyn Homebrew on February 26th and plan on repeating this experiment in the
coming weeks. My presentation will be more streamlined and more time will be given to the students to sample and critique the

beers more data for the experiment. Also, a few posters on homebrewtalk have asked for the presentation slides. I will post them
on February 26th after revising them for my next class. The next experiment will be a characterful English Bitter and I will use an
expressive english ale yeast. I may also increase the over-pitch rate to exacerbate any results. I will also try assess head retention in
each condition. Once the experiment commences I will post here and on homebrewtalk. Hopefully, this will be the last word (at least
for me) on pitching rates.

Pitching Rate Experiment

All homebrewers realize that healthy yeast makes great beer. Our microbial friends need to have adequate oxygen and plenty of
nutrients to ferment a batch of beer to completion without any off-flavors. Another aspect to yeast, besides their health, is the
amount of yeast added to unfermented wort. This is also known as the pitching rate.
This number is important because if you add too little yeast they will tend to grow more, produce more off-flavors such as diacetyl,
and not ferment your beer adequately. Adding too much yeast could lead to yeast autolysis flavors, a thin and watery mouthfeel, and
lack of desired flavors. There is a sweet spot that all brewers (pro and amateur) aim for to excel in fermentation. Pitching rates are
measured in yeast cells per ml per degree of Plato and there are differing opinions as to the ideal number. If you find different
pitching rate numbers the most important thing to do is to determine your own pitching rate on your system as this could vary
between different brewing systems. An average pitching rate that is cited is:
1 million yeast cells / ml of wort / Plato
This number can change depending on several factors:

Ale versus lager. Lagers generally need more yeast to ferment adequately, and the pitching rate could jump to 1.5
million cells/ml/ Plato. Some ales, such as mild ales, may need less.

Gravity. Higher gravity beers need more yeast to ferment the high concentration of sugars (above 1060). Again, 1.5
million cells/ml/ Plato is a good choice for high gravity ales.

Fermentor Geometry. Different vessels can alter how yeast grow and ferment. For example, fermenting in a tall
cylindrical vessel puts more pressure on the yeast so a slightly higher rate is needed. The opposite would be true in a plastic
bucket.

So how do you determine the number of yeast cells to pitch? Lets do some some simple math. Lets say you have a 5 gallons of 1.035
wort and we selected a pitching rate of 0.8 million cells/ml/ Plato. First convert gravity to Plato scale.Divide the last two gravity
points by four to get that number:
Plato = 1.035 gravity / 4
Plato = 8.8
Next, figure out how many millimeters are in 5 gallons of wort. Since 1 gallon has around 3785 mls that would be 18,925 mls. Now
multiply all the numbers together to get your total number of cells needed to ferment the batch of beer:
Cells needed = (pitch rate) * (mls of wort) * ( Plato)
=(800,000) * (18,925) * (8.8)
= 133 billion yeast cells
One important thing I want to mention is the need for a starter. Realizing that we need 133 billion yeast cells for a 5 gallon batch of
low gravity beer, the 100 billion cells provided by WYeast packs, for example, is not enough yeast cells to pitch. Lots of homebrewers
stick to the rule of above a gravity of 1.050 is when you make a starter, but this might not necessarily true and why I always make
a starter for my beer. Go by the numbers and not by conjecture.
=======================
Now this is all well and good, but what does that mean for my beer? Personally, Ive never tasted a beer that was underpitched or
overpitched. Will there be off-flavors? If there are, what kind? What will be the kinetics of fermentation? This sort of experiment has
been done before, but I really wanted to see the results first hand. Moreover, I will be blind tasting the results at a yeast class that I
am teaching at my local homebrew shop, Brooklyn Homebrew.
The experiment will based on the following criteria:

1.

Beer style: Blond ale. I chose a blond ale because any off flavors will have little to hide behind.

2.

Yeast strain: Wyeast 1056. I had a tough time choosing the yeast. I could have gone with a very expressive yeast to detect
changes in flavor. I eventually went with a popular strain that many brewers use for their batches.

3.

Every sample will be treated the same and the test batches will be fermented in 1 gallon batches. The original gravity of the
beer will be 1.046 and only one hop will be used (see recipe below).

Samples:
1.

Control pitch: 0.75 million cells/ml/ Plato 32.9 billion cells.

2.

Overpitch: 2.5 million cells/ml/ Plato 112.8 billion cells.

3.

Underpitch: 0.1 million cells/ml/ Plato 4.7 billion cells.

The recipe (4.5 gallon batch to be split into 1 gallon fermentors):

7 pounds 2-row Pale malt

1 pound of Carapils

Mashed in at 153F for one hour. Raised temps to 168F for mashout and recirculated as usual. First runnings at 1.063. Second
runnings at 1.018. Collected a total of 5.9 gallons at 1.034. Boiled for 60 minutes:

0.5 oz of Centennial at 60 minutes.

0.5 oz of Centennial at 0 minutes (flameout).

Whirlfloc and yeast nutrient at 15 minutes.

Cooled to 62F and pitched the amount of yeast noted above. I did not oxygenate the wort in any sample. Fermented cool (62F)
until fermentation slowed then slowly ramped up to 70F.
On December 14th the class is scheduled and I will post the results, including flavor and fermentation profiles. Im also thinking of
polling the students to see how each beer scored.