I finally got around to doing something I’ve wanted to do since the beginning: bake bread leavened with the yeast isolated from a sourdough culture. A few days before baking I started growing a starter of sorts. I did three steps up from a very small amount of yeast stored in isotonic salt solution. This is similar to what I do for beer but I stopped at about 200mL of weak broth (can’t remember if it was YEPM or MYPG; I forgot to label the jar when I made it). Once I had enough and it had settled I used a sterile pipette to suck up the sediment from the bottom, about 6 grams’ worth of loose, liquid sediment for a 500g dough. It rose overnight in the upper 60′s with a proof of about four hours the next morning. The tiny amount of yeast is in line with what I’d use with dry baker’s yeast for a long rise–it’s probably equivalent to a sixteenth of a teaspoon or so of active dry yeast. The dough is just flour, water, salt and yeast.
I figured it’d not be worthy of a post because it’d probably turn out exactly like a bread made with standard baker’s yeast. After all, it’s just yeast, with no lactic acid bacteria to create the sourness. To a great extent that is true, but the neat thing is that this yeast apparently produces a ton of diacetyl. I had noticed this before when using the sourdough culture it came from (my wheat starter) but I had for some reason assumed it was bacteria doing it. But when I took it out of the proofing basket today I got a massive whiff of butter. This made for a quite nice aroma while baking, although it seems to have completely disappeared in aroma and taste by the time the bread cooled.
These yeasts were the fastest growing (at least in terms of visible colonies) that I’ve seen so far. I suppose that’s consistent with what you’d expect from a continuously-refreshed culture like sourdough, but that’s just speculation. I have no idea what species they are, but I’m fairly certain it’s not S. cerevisiae based on the quick growth and colony characteristics. The colonies spread out much more than I’ve seen with S. cerevisiae. They are also whiter and flatter. I keep meaning to ferment wort with it to see its alcohol tolerance and sugar utilization but never get around to it.
This is the third and final attempt at plating out the whole sourdough starter. I did a few things differently:
- I fed the three sourdough starters and let them ferment for 12 hours before starting. Last time two of the starters were right out of the fridge.
- I put even less of each culture in the sterile water to dilute it
- I used bromocresol green + potato dextrose plates, plus one potato lactose plate
The rye starter’s plate grew mold in a few days. It totally took over before I had a chance to see anything else growing. The other two starter’s plates plus the lactose plate (on which I plated the wheat starter) were totally fine. I let them grow out for 16 days.
Clockwise from top left: brown rice flour starter, wheat flour starter, wheat starter on potato lactose plate
The colonies look translucent and shiny from certain lighting angles, but they are off-white, smooth and round and actually matte. I did not see any flat colonies like last time. The weirdest surprise is the almost identical growth on the lactose plate. I am going to assume that this is due to whatever sugar was in the potato and not the lactose. My goal in doing so was to see if I could get the bacteria to grow on there and was not expecting the yeast to grow at all. You can see very tiny colonies on each of the plates between the large yeast colonies and smaller yeast colonies. They are transparent and shiny. I believe these are the lactic bacteria.
I think I may not have used enough bromocresol green, or the plates were old enough that it had already started to fade. I was expecting green colonies here, too, but they were white. On my first (recent) attempt I did see green colonies but the plates were also darker back then. The colonies looked quite similar between the two starters. This is a change from the first time I did this (earlier in the year) but could be because I am using a different medium.
I picked out a yeast colony from each and plated it on YEPM medium (recipe from jaapie’s site; the ingredients are not cheap or easy to find for hobbyists like me). In the past, the yeast I isolated from the wheat starter didn’t appear to ferment maltose whereas the rice starter’s did. We’ll see if these grow on it (from what I can tell, yeasts generally but not always can grow on something they can ferment and vice-versa).
On another note, I finally resumed the DCambic project from April. I had isolated a yeast from it back then but it appeared to not do much besides make lots of strong fruit esters. Notably, it didn’t seem to produce any alcohol. This time I instead went for growing it in liquid first instead of plating directly from the sample. I now seem to have two different types of yeast in liquid, judging from the two colors of sediment I see. A darker sediment formed first. I then started shaking to aerate the culture on a regular basis for a few days and then added this to more sterile wort, which I similarly shake on a regular basis. I am now seeing a lighter color sediment forming as well. I also see lots of evidence of fermentation, though I did not see any krausen at any point. I shook up the culture yesterday and streaked a sample on a PDA plate. I’m hopeful that this time I can get the yeast that produces the distinct Bretty aroma I can smell from the culture.
These are the results after 5 days of growth.
Wheat flour starter (Carl’s Friends) on top. Bottom left is the brown rice flour starter (homegrown) and the right is the immature rye flour starter.
I am a bit surprised for a few reasons. First, from looking at them I was convinced that there was no yeast growth. At first I had all three of them in a big plastic bag, and the smell was not at all yeasty. It also looked like all of the colonies on the wheat and brown rice starters were very glossy like bacterial colonies. I had previously plated both of those on malt extract agar and the yeast colonies looked very different from each other, but definitely looked like yeast. I saw nothing of the sort here. The rye starter definitely had nothing that looked like yeast, but it is still young.
The first surprise came when I opened the tops to photograph them. I first took some of the colonies from the wheat starter plate and put them in a bit of light wort. But when I took a whiff both the wheat starter plate and rice starter plate smelled very yeasty. The smell from the bag I mentioned before was from the rye starter plate. It smelled not wholly unpleasant. I could pick out acetic acid but there were other things underneath. The colonies were tiny and slightly yellow. I’m not sure if they were heterofermentative LAB that produce acetic acid or some sort of enteric bacteria.
In the end I tossed out all the plates. But now looking at the picture above I can see that the colonies aren’t equally glossy. On malt extract agar (a long time ago) the wheat starter’s yeast were flat, dull and bigger than brewer’s yeast. That kind of matches some of the colonies I see in the picture. The rice starter’s yeast looked and behaved a lot like brewer’s yeast (I planned on trying it for brewing but never got around to it. I just made some small liquid cultures.) I don’t see anything of the sort here.
Later today I’m going for a third try on the bromocresol green and dextrose plates. Hopefully I can get some distinct yeast growth since LAB aren’t supposed to grow on it.
The sourdough stuff I promised in the last post hasn’t gone too well so far. I made the plates I mentioned but doing things with them has not gone well.
The idea is to do a few things:
- Compare the organisms in three sourdough starters
- Evaluate the properties of the yeasts I can isolate
- Try to learn a thing or two about lactic bacteria
- Try to cultivate a new starter (the third from above) from scratch, which I hadn’t done in a while
So the fourth bullet is sort of working, but not without some problems. I made an effort to use sanitized glasses and silverware to handle the starter so that I could be reasonably sure the organisms were coming from the flour and not from my environment. It took a long time but I can’t say whether that is due to that or some other reason. After a lot of strange stages (producing what I think is DMS, since it smelled a lot like cooked corn) it looks like it is becoming a viable starter. However, I messed up one day and stuck an unsanitized fork in there. It was just starting to turn around, but it sped up its turnaround after that. Having recently worked with other starters it could be I “contaminated” it with known-good organisms.
(As a side note, I always wonder if the number of environmental bugs is sufficient to overcome whatever is in the flour, but I suspect that is highly complicated and not subject to a simple yes/no assessment. I wish I had access to irradiated flour so I could put it to some sort of test.)
The first three require me to plate the starters. My first effort at this was destined to fail: I tried to streak after dipping a needle in the starters. I got some bugs from the early 2nd day stage of my new starter, some bacteria I can’t identify (yellow colonies on PDA). The other plates grew only a handful of yeast colonies but the plates grew tons of mold before I could do anything else with them. In any case, the bits of liquid dough along the streaks made it quite difficult to tell if things were growing at first.
I repeated the effort this unplanned long weekend, since I was home from the storm. I went with spreading a sample of sterile water that had a bit of starter dipped in it. After 24 hours there is no growth at 75F. We’ll see what happens in the next couple of days.
Despite the total lack of posting, I am not anywhere near done with the blog. As I mentioned in the last post I had a three week trip followed by moving, and it took until the end of August to settle down. Plus, I also got engaged.
I am preparing some potato plates today or tomorrow. I have an idea for some posts about sourdough. So, stay tuned!
As you may have guessed from my posting frequency, I’ve not done much with yeasts recently. A flurry of unfortunate events in the past month has kept me busy, and with an upcoming international trip followed by a move when I get back there will be little going on until August or so. So, I will leave you with the only interesting thing happening lately, which is that the sample I inoculated with a few drops of the DCambic grew a pellicle.
It’s actually grown two pellicles now. The first fell when I moved the bottle, disappearing completely in a few hours. This one grew a week or so later and also fell, but not before covering the entire surface. The neat thing is that you can see the sunken pellicles at the bottom of the flask.
In addition to the pellicle, I’ve also noted a few things. First, there is a layer of dark sediment which I also noticed in the vial Mike gave me. Some sort of yeast, perhaps, but I have no way of knowing right now. Second, about a week or so before the first pellicle showed up I started seeing CO2 bubbles, which I had not seen before. The yeast ring around the flask came from that stage. All of this almost two months after the initial inoculation. I will have to repeat this after I move and then try to isolate the bugs from there instead of from the vial like I did the first time.
It looks like nothing else is going to grow on the plates, so I went ahead and streaked out one of the colonies on another plate and inoculated a small liquid sample. I will step that up as usual. The plate had a slightly funky smell unlike any yeast plates I’ve dealt with so far. The colonies also look bigger, glossier and more irregular than others I have dealt with. I have not yet worked with laboratory Bretts, though, so I can’t compare. In any case, given that the previous liquid sample grown from the raw beer developed an intense Bretty character I am quite sure that is what I’m dealing with here. I’m not too good at describing the various Brett flavors, so I’ll venture only that it is fruity and phenolic. Although there was no visible fermentation there is definitely alcohol in the aroma. I am imagining the new liquid sample from the pure culture will smell the same.
I had originally planned to streak out two plates, one with potato dextrose and one with potato lactose, but I ended up not having the time to prepare them this weekend. I went instead with another malt extract plate. This doesn’t help me get any new information on how the strain behaves but it’ll do for now. When I have the time I will prepare those potato plates with bromocresol green and restreak. One nice thing about potato media is that you can easily swap the sugars and make different types from the same batch of potato infusion.
Nothing at all grew on the brilliant green plate. My previous experiments with them had at least some yeast growth but it was very slow. This time I don’t see anything. It is safe to say my experiment with that medium is a failure. Good thing it was unnecessary for this experiment.