Anyone know where to obtain beta amylase enzyme (without any other enzymes mixed in) for a reasonable price? I have an experiment I would like to do, but it wouldn't be complete if I couldn't get isolated beta amylase.
Brew o
Brew o
Thanks, but I've seen that listing. I'm not interested enough in the experiment to spend $300+ to run the experiment.
This is a lot more reasonable. Thank you.Not sure what your budget is, but Fisher has 25g for $69.
https://www.fishersci.com/shop/products/beta-amylase-soybean-tci-america/A044825G
Comments? @VikeMan
Very interesting! My suggestions, some of which you are probably already planning, but omitted for brevity:
- Boil or otherwise sanitize/sterilize the worts before fermentation. Yes
- Measure the OGs. Yes
- Use a yeast strain that's non-diastatic. Of course, no Belle Saison here. Probably use US05.
- Consider repeating (if affordable) with one hour mashes and no cereal mash, to approximate normal/typical mash conditions. Could remove possible objections that could arise from only doing it with a cereal mash and longer mash. (I wouldn't personally predict the basic result to be different between the two trials, but I'd still probably do them.) A shorter mash series would be interesting to get an idea of how fast the enzymes work, and at different temps too. If the cereal mash isn't done, then at lower temps both beta and limit dextrinase will be significant, with no way to separate out the effects of each.
- If planning to pulverize the grains, do a dry run to make sure they don't tend to "brick" at the bottom of the mash vessel. I've seen that with some malts during pH experiments. Plan is to use my normal BIAB crush @ 0.022" gap on my two roller mill.
ETA: - Consider repeating with different total enzyme concentrations. We'll see how things go.
There are lots of useful enzymes and mix combos available. But for the purposes of this experiment, I need to specifically have beta amylase, and this is not commonly available.I just purchased a vial of enzymes designed for the fermenter, to make a brut IPA/Lager. The LHBS had three different compounds, and I just bought the one he suggested, but I have no idea at the moment what % of which enzymes are in it or the recipes for the ones I didn't buy (one of which was mash-oriented). Are these compounds that difficult to find or is it that your parameters were very specific?
This is what I love about HB and this website. We science nerds love these ideas.I might as well go ahead and outline the experiment I am looking at.
It is my belief, based on research, that it is really limit dextrinase, and not beta amylase, that is responsible for the higher fermentability of worts mashed at low initial temperatures compared to worts mashed at higher initial temperatures. So, I have an experiment in mind that could confirm or refute this hypothesis.
The basic outline is:
If the wort mashed with beta + alpha has a lower FG than the wort mashed only with alpha, then beta amylase action does in fact lead to higher fermentability than mashing with alpha alone. If the two worts finish at the same FG, then beta action does not actually do anything to increase the fermentability of worts. If the second case holds, then the higher fermentability of low mashed worts is due to something other than beta amylase.
- Denature all of the endogenous enzymes by doughing in to boiling water, and then conducting a cereal mash to fully gelatinize all of the starch in the grain.
- Cool the mash down to ~145°F (63°C) and maintain temp with a Sous Vide controller.
- Add either alpha amylase alone, or alpha + beta amylase to the mash, and let the mash go for ~2 hours to make absolutely sure that the enzymes have done all they possible can.
- Ferment both worts using a fast ferment test schedule (with the same yeast obviously.)
Comments? @VikeMan
Brew on
Control would be a mash conducted normally at the ~145°F temp. If my hypothesis is correct, this wort would have higher fermentability, due to the presence of limit dextrinase, than either of the experimental cells.This is what I love about HB and this website. We science nerds love these ideas.
What do you propose for a control? Are you going to split a batch with and without the enzyme from the same boil? Keep us posted with your progress.
I think this is pretty much the only way to make a real Brut. Diastatic strains make their own glucoamylase, but I don't believe that they'll ferment down to 1.000 or below.If I was doing a super dry Brut I would use glucoamylase as that will rip virtually everything and end up at 1.000 fg...
I guess you can't really do the perfect experiment since it's pretty much impossible to buy limit dextrinase. I suppose you could try making your own.Control would be an mash conducted normally at the ~145°F temp. If my hypothesis is correct, this wort would have higher fermentability, due to the presence of limit dextrinase, than either of the experimental cells.
The extraction method is a little beyond my means, both equipment and skills. Will have to stick to the simple experiment.I guess you can't really do the perfect experiment since it's pretty much impossible to buy limit dextrinase. I suppose you could try making your own.
Is your working hypothesis that limit dextrinase contributes to wort fermentability, especially in low-temperature mashes, or is it that beta amylase does not have a role? I don't think the first statement is at all controversial.
If it's the second, it would be interesting if you could provide more of your reasoning.
In particular I'm wondering about high-temperature (75 C) mashing to produce wort with poor fermentability (for the low-alcohol folks), where the standard explanation is that you're eliminating beta amylase activity and as a result leaving behind longer 1,4-linked oligosaccharides.
I never postulated that beta amylase does not contribute to starch hydrolysis. It does, and it makes the overall process of saccharification go faster. But since gelatinization is the rate limiting step in starch conversion, the speed of hydrolysis is a minor factor in time to "complete" a mash. What the experiment is designed to elucidate, is whether or not having beta working with alpha results in more fermentable wort - so, just to support your so called "weaker conclusion", and that beta is not responsible for the higher fermentability of worts initially mashed at low termperatures. To do this, we need to denature the limit dextrinase very early on in the mash, unfortunately, this will also denature the beta and alpha amylase, thus the need to add exogenous amylase enzymes for the experiment.It seems like your experiment will have to be careful to match the enzyme concentration and duration of a typical mash. Otherwise, you will only reach a weaker conclusion: "Given enough alpha amylase and enough time, you can achieve a similar fermentability without beta amylase," instead of "In a normal mash, beta activity does not contribute to starch hydrolysis."
Indeed, I would expect this to come pretty close to reality. Not sure what a diastaticus yeast like 3711 or Belle might be able to do here though.If alpha amylase alone is sufficient to get you all the way to limit dextrins, you’d expect an alpha-only mash to be ~85% fermentable sugars.
I am not certain but have the feeling that this the very low end of the fermentability range for any hot mash of any starch. Even bread & water with no added sugars will somehow ferment to about 50% AA. Ever heard of kvass? It is one of the strangest "beers" I know of, fermented bread-water. I made it once. Some of the starch and dextrins seem to be able to be broken down enough via whatever process (I'm not certain how) to allow partial fermentability. There's probably just too many variables for me to be able to understand how this works -- a little sugar in the bread, a little enzymatic action within the yeast cells, the temperatures involved breaking down some dextrins into actual fermentable sugars... boggles my mind somewhat that yeast is still able to eat a large percentage of what should be not far from 100% complex starch.High-temperature mashing gives you wort that’s <50% fermentable.
Ferment everything, all the way down. The glucoamylase produced by diastatic yeasts will break all starch down into simple sugars. (Non-starch carbohydrates, like beta glucans, are not broken down.)Indeed, I would expect this to come pretty close to reality. Not sure what a diastaticus yeast like 3711 or Belle might be able to do here though.
The last beer I made mashed at 167 F went from 1.025 to 1.016 for 36% apparent attenuation.I am not certain but have the feeling that this the very low end of the fermentability range for any hot mash of any starch. Even bread & water with no added sugars will somehow ferment to about 50% AA. Ever heard of kvass? It is one of the strangest "beers" I know of, fermented bread-water. I made it once. Some of the starch and dextrins seem to be able to be broken down enough via whatever process (I'm not certain how) to allow partial fermentability. There's probably just too many variables for me to be able to understand how this works -- a little sugar in the bread, a little enzymatic action within the yeast cells, the temperatures involved breaking down some dextrins into actual fermentable sugars... boggles my mind somewhat that yeast is still able to eat a large percentage of what should be not far from 100% complex starch.
For a hot mash in the zone for alpha only, where the beta (and I suppose the limit dextrinase also) have been denatured, I have figured one can obtain a reasonably fermentable wort anyway simply by mashing for a long time, such as with an overnight mash of 6-10 hours or whatever. I've yet to experiment with this hypothesis.
If this were the case, then boiling wort should work even faster to hydrolyze starch to sugar, since the temp is higher, and reactions go faster the higher the temperature (all else being equal.)Will there be spontaneous breakdown of the sugars in the warm Liquor?
The enzyme just catalyses the reaction to get the conversion in a suitable time I suppose.
The last beer I made mashed at 167 F went from 1.025 to 1.016 for 36% apparent attenuation.
But then, wort extract is only about 90% carbohydrates, so if 85% of that 90% is fermentable, then the fermentability is about 76-77%. Apparent attenuation will be different than that since the presence of alcohol reduces the SG.The estimates I’ve found are that once the amylolytic enzymes have broken all the 1,4 bonds they can, the little 1,6-branched pieces that are left comprise about 10-15% of the total starch.
If alpha amylase alone is sufficient to get you all the way to limit dextrins, you’d expect an alpha-only mash to be ~85% fermentable sugars. High-temperature mashing gives you wort that’s <50% fermentable.
Isn’t it beano?Anyone know where to obtain beta amylase enzyme (without any other enzymes mixed in) for a reasonable price? I have an experiment I would like to do, but it wouldn't be complete if I couldn't get isolated beta amylase.
Brew o
Isn’t it beano?
Reddit...Only said it because someone recommended it on Reddit for a too-high mash
if you want really dry, use amyloglucosidase (aka glucoamylase or just gluco) in the fermenter.
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