Cantillon microbiota

[suregork]

Just thought I'd link to an interesting (and open access!) study that was published last week on how the microbiota of two batches of Cantillon lambic develop over a two-year period:

http://www.plosone.org/article/info:doi/10.1371/journal.pone.0095384

Abstract

Lambic sour beers are the products of a spontaneous fermentation that lasts for one to three years before bottling. The present study determined the microbiota involved in the fermentation of lambic beers by sampling two fermentation batches during two years in the most traditional lambic brewery of Belgium, using culture-dependent and culture-independent methods. From 14 samples per fermentation, over 2000 bacterial and yeast isolates were obtained and identified. Although minor variations in the microbiota between casks and batches and a considerable species diversity were found, a characteristic microbial succession was identified. This succession started with a dominance of Enterobacteriaceae in the first month, which were replaced at 2 months by Pediococcus damnosus and Saccharomyces spp., the latter being replaced by Dekkera bruxellensis at 6 months fermentation duration.

I apologize if this has been posted already somewhere in the forum

 

[bestbuds]

Thanks so much for posting this! If anyone has any questions about the scientific methods used, I'd be happy to try to answer.

Interesting that Brett doesn't really show up in big numbers until 6 months into fermentation. It's also interesting that Pediococcus damnosus is so prevalent - this species has its name because of the excessive amount of diacetyl it is capable of producing. I'm wondering if other species in the ferment are able to reduce or take up the diacetyl in a more efficient manner than Saccharomyces - I don't think many people complain about diacetyl in Cantillon beers.

 

[bestbuds]

It would be interesting to do a controlled lambic following the same timeline - Hanseniaspora (and avoid the enteric bacteria) at the start, then Saccharomyces, then Lacto/Pedio and Acetobacter, finishing up with Brett and more aging.

 

[Johnnyhitch1]

So.....awesome

Thx for the post!

 

[dcp27]

Thanks so much for posting this! If anyone has any questions about the scientific methods used, I'd be happy to try to answer.

Interesting that Brett doesn't really show up in big numbers until 6 months into fermentation. It's also interesting that Pediococcus damnosus is so prevalent - this species has its name because of the excessive amount of diacetyl it is capable of producing. I'm wondering if other species in the ferment are able to reduce or take up the diacetyl in a more efficient manner than Saccharomyces - I don't think many people complain about diacetyl in Cantillon beers.

brett cleans up diacetyl

 

[bestbuds]

brett cleans up diacetyl

Can you please link to the source of the info? I believe you, just want to get more information!

 

[Fermented_minds]

Can you please link to the source of the info? I believe you, just want to get more information!

Both Brettanomyces and Saccaromyces have the enzymes needed to reduce diacetyl to acetoin, and then to 2,3 butanediol:

http://pathway.yeastgenome.org/YEAS...bject=PWY3O-981&detail-level=3&detail-level=2

Can't find the super pathway for Dekkera/Brettanomyces, but there is significant overlap in genetic sequences for Saccharomyces and Dekkera/Brettanomyces that serve as the code for diacetyl reductase and 2,3 butanediol dehydrogenase enzymes. You can find that information by simply clicking on the enzyme in question and running a blast comparison to compare the Saccharomyces to other species including Brettanomyes(Dekkera).

 

[bestbuds]

Yes, but Saccharomyces (and presumably Brett) are not able to clean up the amount of diacetyl produced by many Pediococcus species - which can be several fold higher than that produced by even the butteriest Saccharomyces. It would be really interesting and potentially extremely useful in the industry if it turns out Brett uses that pathway more effectively, allowing it to reduce higher concentrations of diacetyl, however. Hmm.

 

[Fermented_minds]

Yes, but Saccharomyces (and presumably Brett) are not able to clean up the amount of diacetyl produced by Pediococcus - which can be several fold higher than that produced by even the butteriest Saccharomyces.

I have read this before on this very forum, and nary has a source been cited for it. If you have one, I'd be interested to read over it, but it appears to be completely contrary to the experience of many that use Pediococcus with Brettanomyces and, to a lesser extent, Saccharomyces. Not sure there is much hard research on this, though there should be.

Diacetyl coming into the cell is a passive diffusion process dictated by the concentration of diacetyl both in the cell and in the external environment. Its reduction by the requisite enzymes is also concentration dependent with respect the the concentration of enzymes, the concentration of reactant (diacetyl) and the concentration of products (acetoin and 2,3 butanediol). The simple laws governing how these things work would indicate that the presence of increased diacetyl will drive diacetyl (reactant) into the cell, which will then drive the reduction to acetoin and then 2,3 butanediol (products).

It may be the case that different yeast make different quantities of the required enzymes and that is the rate limiting factor. However, in that case a longer period of time will simply be required for the diacetyl to be reduced in a given beer by a yeast that produces lower levels as opposed to one that produces higher levels.

I'd love to see more focused research on this, with some comparative analysis between different yeast species and some solid enzyme kinetics experiments! There's a paper just waiting for someone to do the research and write it up.

 

[sweetcell]

This won't satisfy the requests for a scientific source, but both Vinnie and Jamil have unequivocally stated that one must use Brett with Pedio in order to get rid of Pedio's diacetyl. I trust these guys' opinion on yeast and sour beer fermentation.

The real proof of sorts, for me, is that there are no beers that contain Pedio that do not also have Brett. Why are there no Sacch + Pedio (only) beers? There are Sacch + Lacto beers...

Yes, but Saccharomyces (and presumably Brett) are not able to clean up the amount of diacetyl produced by many Pediococcus species

What is your source for this statement? Or maybe you have experienced this? I was under the impression that in the short term, Pedio will overwhelm the yeasts but given enough time Brett will clean it up.

 

[bestbuds]

This won't satisfy the requests for a scientific source, but both Vinnie and Jamil have unequivocally stated that one must use Brett with Pedio in order to get rid of Pedio's diacetyl. I trust these guys' opinion on yeast and sour beer fermentation.

The real proof of sorts, for me, is that there are no beers that contain Pedio they do not also have Brett. Why are there no Sacch + Pedio (only) beers? There are Sacch + Lacto beers...


What is your source for this statement? Or maybe you have experienced this? I was under the impression that in the short term, Pedio will overwhelm the yeasts but given enough time Brett will clean it up.

You're right, anecdotally it seems to be true but I can't find public research backing up the phenomenon.

I was taught in a brewing class that Saccharomyces has a maximum concentration of diacetyl that it is able to clean up, but I don't think I was provided an exact amount. Furthermore, I can't find this information in any brewing books or Pubmed. Hmm.

 

[suregork]

Here is a recent study, where up to 50 mg/L diacetyl (about 50 times what is a typical peak concentration of 'total diacetyl' (alpha-acetolactate + free diacetyl) in wort during fermentations with Saccharomyces) was supplemented to grape must prior to pitching of yeast. As you can see from Figure 1, despite the high initial concentrations of diacetyl, it was rapidly reduced during fermentation. It is interesting though that the final diacetyl concentration in the wine produced from the supplemented must (~2 ppm) was quite a lot higher than the control must (~0.4 ppm).

http://onlinelibrary.wiley.com/doi/10.1002/jib.106/abstract

 

[Fermented_minds]

Here is a recent study, where up to 50 mg/L diacetyl (about 50 times what is a typical peak concentration of 'total diacetyl' (alpha-acetolactate + free diacetyl) in wort during fermentations with Saccharomyces) was supplemented to grape must prior to pitching of yeast. As you can see from Figure 1, despite the high initial concentrations of diacetyl, it was rapidly reduced during fermentation. It is interesting though that the final diacetyl concentration in the wine produced from the supplemented must (~2 ppm) was quite a lot higher than the control must (~0.4 ppm).

http://onlinelibrary.wiley.com/doi/10.1002/jib.106/abstract

Wonderful find, suregork. I think that fact that there is SOME research on this is good. Thanks for sharing!

I have always been of the opinion Saccharomyces could likely clean up copious amounts of diacetyl, as I have used Pedio without Brett and the beer was great, no butter. It was a Pale Ale I soured, and since the IBU were a little higher I tossed in Pedio AND Lacto in hopes it would still sour. I primaried with trusty 1056. A word of caution on my anecdotal evidence though, Pedio may have not created that much to begin with and even if it did and the Sacch cleaned it up, it may be strain dependent as I theorized earlier.

 

[sweetcell]

as I have used Pedio without Brett and the beer was great, no butter.

aha! we've found one!

 

[bestbuds]

Here is a recent study, where up to 50 mg/L diacetyl (about 50 times what is a typical peak concentration of 'total diacetyl' (alpha-acetolactate + free diacetyl) in wort during fermentations with Saccharomyces) was supplemented to grape must prior to pitching of yeast. As you can see from Figure 1, despite the high initial concentrations of diacetyl, it was rapidly reduced during fermentation. It is interesting though that the final diacetyl concentration in the wine produced from the supplemented must (~2 ppm) was quite a lot higher than the control must (~0.4 ppm).

http://onlinelibrary.wiley.com/doi/10.1002/jib.106/abstract

Awesome find! Coauthored by a former advisor of mine even.

 

[mot]

Can you please link to the source of the info? I believe you, just want to get more information!

You will have numerous talks and hits with a google search on that topic

 

[mot]

Thanks so much for posting this! If anyone has any questions about the scientific methods used, I'd be happy to try to answer.

Interesting that Brett doesn't really show up in big numbers until 6 months into fermentation. It's also interesting that Pediococcus damnosus is so prevalent - this species has its name because of the excessive amount of diacetyl it is capable of producing. I'm wondering if other species in the ferment are able to reduce or take up the diacetyl in a more efficient manner than Saccharomyces - I don't think many people complain about diacetyl in Cantillon beers.

Brett's pretty slow growing, what makes more of a surprise to me is the large amount of lager yeast

 

[tootal]

My friend and I just returned from a 2 week Euro beer tour which included the zythos festival in Leuven and Night of the great thrist south of Brussells. During our visit we went to the Cantillon brewery and got to talk to the owner/brewmaster. He has good news as he is getting another building down the street to store,(lager), his wooden barrels in. The reason he can only brew so much is he runs out of space for the barrels. Now he will be able to brew longer and increase production. Of course, since it takes three years to make Geuze we won't notice anything for a while!

Very cool place by the way.

 

[smokinghole]

I think when it comes to research available for diacetyl reduction by yeast the best you'll find is a wine industry based research. Since the wine industry is backing the study that's what they'll study. You will not likely find research profiling diacetyl reduction ability of brett or dekkera because the wine industry hates them and don't care how well they reduce diacetyl because they don't want it in their musts. Although there is a realization that some regional styles achieve their signature flavors because of brett/dekkera. Joke's on the snooty brett/dekkera haters.

Anyhow research showing the how and why isn't a concern of mine. I've yet to drink a beer with pedio that also had brett that had any sign of diacetyl. Given enough time I would wager that it would be reduced no matter how productive the pedio is at making diacetyl. I've actually thought about using sausage making strains of pedio in beer since they're intended for fast growth and acid production.