Official Enzyme Mashing Method Thread

[Dan Call]

Enzymes seem like hit and miss. I'm on my fourth full blown mash with them and it's starting to make a little sense as the sweetness and liquification is getting better. I would like for us to compare notes on this. There seem to be a few things that are absolutely crucial to getting them to work. To keep things simple, it's just all corn, cracked corn, and genuine spring water with no treatment. Using Superstart yeast, it's very sturdy yeast and won't quit. Enzymes are alpha and gluco bought from Mile-Hi. Throwing in a little DAP. Using citric for downward adjustment. Start off the mashes with the 5.2 adjuster.

1. Cook the p*ss out of the corn.
2. See #1
3. Adjust PH down to around 4 before adding alpha. (digital PH very handy)
4. Pre-malt with alpha at somewhere around 170-180 - not getting substantial liquification.
5. Alpha works best at less than 160.
6. Gluco works best at around 140.

One thing I've been looking for but not getting is that clear syrupy layer on top like with malted barley. Nevertheless, I'm getting great liquification. One thing that's helped is sealing off the top of the cooker and letting it sit for a while and just steep, this really seems to have an effect.

30 gallon cooker. Starting off with 10 gallons water, 30 pounds cracked corn. Let water be in rolling boil before adding corn. Stir like crazy. After it reaches the porridge stage, let sit sealed for a while, add five gallons of water, brings down temp to aroun 160, add alpha, really things up, add gluco really really thins up, let sit. This last one has liquified on the top more than any of the other four.

The only real difficulty I'm having, besides conversion, is getting a proper SG reading. The mashes liquify so much I think it's showing artifically low SG. I can't figure this out. I know how to effect and get proper SG. It's just that these mashes are thinning out so much it seems to make the SG goofy. But they are fermeting their nuts off.

This is all I know....so I'm told. So I think.

[Dnderhead]

the pre malt/enzymes is to keep your mash from getting to thick to stir,not to convert. with that mash you should be getting approxmently OG 1.054
FG 1.014.--abv 5.3,, even if it does not fully convert , the enzymes will keep working if you do on the grain ferments.yeast and enzymes will work together.

[blind drunk]

So the PH is important at this stage as well? I didn't get the full conversion but its fermenting like hell and smells as sweet.

[1fourme]

1. don't need DAP, 5.2 will adjust water.
2. drop your temp to 160 for pre malt, I normally heat to 160 turn off heat, then add corn, it drops temp to 145-150 then add either small amount malted barley and cover and wait 20-30 min. or add alpha and wait.
3. boil 1 hour turn off and cool to 140 then add gluco. I leave covered overnight and add yeast in the morning,
temp over 160 kills enzymes.

[Dnderhead]

The grain will alter the PH . and yes enzymes work best at about ph 4-5 same as yeast. grain will bring it down close to that . most of this is they are
reading is working with beer.and what is optimal. not so with distilled beverages.
this is not necessary with what we do. also with beer you want fast conversion, lower the temps fast to keep out off flavors and contamination.
something we are not worried about. as you know yeast will work at lower temps. just takes longer enzymes the same way.
do you check your bread for PH ? analyze the water for minerals and PH?
I do a lot of grain mash, use well water. cook grains add malt, then yeast and I git about 80% conversion.unless you have soft water
( in that case mite want to add calcium) you do not have to worry about it.

[dixiedrifter]

Try grinding your corn to the consistancy of cornmeal. That increases the ability of the grain to absorb water and geletanize the starches.

When initially adding in the alpha-amylase, the pH of the mash should be around 5.8 to 6.0 for maximum activity. A pH below 5.2 or above 7 will produce less than desirable results.

Think of enzymes as having a half life like uranium or other radioactive materials. After x amount of time, half of the enzymes will become denatured, with another half being denatured in another x amount of time till there is very little working enzymes left or the starch has been fully converted. The half life rate depends on temperature, pH, and time.

Generally increasing the temperature increases the activity of the enzyme exponentially while the half life more or less gently decreases provided all other variables are optimum. This is why we mash at higher temps rather than dumping them in at room temp.

Bacterial based alpha-amylase enzymes are somewhat to greatly more thermostable than those derived from malted barley or fungal amylases. This is because all enzymes are essentially large proteins and each species or strain used in their manufacture produces a slightly different enzyme even though they do the same thing. As such, different manufacturers use different microbes and their enzymes will have different optimium parameters. Non-thermostable bacterial alpha-amylase generally shows activity up to 85ºC before it begins to denature with peak activity around 65-70ºC, or around the point where the starch gelatanizes. If it were me, I'd shoot for a 70ºC strike temp and see how it goes. If it works, then kick it up to to 75ºC and finally 80º. Remember, your strike water takes a temperature of approx 10ºC more degrees greater than your target strike temp depending on how much heat your mash tun absorbs upon addition of strike water.

Alpha-amylase should have some free calcium ions in the solution. This helps to stabilize the enzyme and make it work more efficiantly. 2 grams per gallon of anhydrous calcium chloride per gallon of should be sufficiant to accomplish this. That should result in a Ca++ ion content of around 144ppm. Bumping it up to 3 grams per gallon would result in a level of 216ppm. In practical terms, calcium chloride normally comes in the form of a dihydrate so 3 grams should result in somewhere between 144 and 216ppm. Be aware that it is possible to overdose on the calcium chloride which results in the amylase being denatured.

Alpha-amylase should be added in two doses. The first dose is for liquefication purposes during cooking and the second is where the actual dextrinization occurs. One could theoretically add strike water at the proper temperature to achieve a mash temp of about 80 degrees or so, then let it sit in a cooler and cook till the temps drop to ideal temps for the second addition. This will eliminate scorching issues.

When cooking other enzymes may be added to assist in liquefication. One such enzyme is Phytase which will work to neutralize phytic acid which is detrimental to alpha-amylase. Supposedly it also allows alpha to work at a lower temperature. Here is one site that might be useful source of this and other enzymes... http://nutriteck.ca/sslshop/product_inf ... cts_id=145" onclick="window.open(this.href);return false;" rel="nofollow

Mash grain to water ratio comes into play as well. A thicker initial mash helps to stabilize the alpha-amylase enzyme. Industry uses a 30% or so average w/w grain to water ratio. Some references are vauge as to whether the ratio includes or does not include the moisture content of the grain when calculating the ratio. 1 gallon 30% slurry by volume taking consideration of the latent moisture in the grain should consist of approximately 1425g of corn with the remainder being 2739mls water. Converted into pounds/quarts thats 3.13 lbs corn to 2.85 quarts of water or a water to grain ratio of .91 Using plain corn weight, those figures changes to 1250g corn and 2932 mls water or 2.75lbs corn to 3.1 quarts water needed to make one gallon of slurry volume. Remember, you'll use slightly more grain/water than the weight of one gallon of water since corn is more dense than water. In any case a simple 1.25 ratio water to grain ratio should be acceptable which would be 2.5lbs grain to 3.2 quarts of water to make one gallon of mash by volume.

Fortunately glucoamylase or amylogluconase or GA-100 is much simpler to use since it can be used in a simultaneous saccarification and fermentation process. Think whiskey yeast with AG. The only catch is glucoamylase requires a lower pH than malt barley or alpha amylase. Peak activity is normally around pH 4.3 however this is dangerously close to too low a pH for ideal yeast activity. An ideal pH for a mash would be around 4.6. Several acids can be used to accomplish this with industy using hydrochloric or sulfuric acids. In addition, pH buffers can be used such as acetic acid/sodium acetate or citric acid/sodium citrate. The pH buffer sold in 1lb jars to set the pH at 5.2 should not be used with synthetic enzymes since it is too high for the glucoamylase to work properly, however, it is perfect for malted barley.

[Dan Call]

Goodness what diverse, yet informative, comments.

I'm not using DAP for PH adjustment, just a little bit extra yeast help.

We seem to have wildly divergent cook-pre-malt methods here. But this is why I started the thread. To get an idea of a methodical way to do it.

I've done some sugar things before, but am strictly all grain now.

Please, keep it coming.....

[T-type]

So do you usually boil corn for 1-1/2 hour or so and then let cool to 160f, pitch alpha and wait 30 minutes and then pitch gluco? and let sit all night? or is are you suppose to pitch alil alpha before you start boiling so it doesnt get so thick? This is what i got from reading anther thread:
"Re: Enough Enzymes ??
by Dnderhead on Sun Mar 15, 2009 11:48 pm

What I do is mash so I can leave it for a while (usually takes me all day any way) but I cook ,cool, add malt and stir, turn off all heat,
rap in old blanket or something to hold heat, an just leave over night at room temps. (some use water cooler)"
Im guessing i can substitute this with alpha at 160 and let sit then add gluco at 140 and let sit all night.

Or do you like to start with your corn and water in pot not yet boiling and at a temp of 160 or so and hold it there with your alpha for a hour or so, then raise temp to boiling and boil awhile. then let cool to 160 and throw in your gluco and let sit covered all night. in the morning transfer to fermenter and pitch yeast.
Im just confused on different methods i hear about.

[punkin]

Just to throw another cat on the fire, my alpha is optimised for 100 C temps. ie, boiling temp.

I'm with DD, the spec sheets for my (liquid) enzymes say that the alpha likes a ph round six and the gluco likes a ph under 5. This is easy to do by lowering ph just before the addition of gluco.

[dixiedrifter]

Just to throw another cat on the fire, my alpha is optimised for 100 C temps. ie, boiling temp.

I'm with DD, the spec sheets for my (liquid) enzymes say that the alpha likes a ph round six and the gluco likes a ph under 5. This is easy to do by lowering ph just before the addition of gluco.

Right. If I were in punkin's shoes I would simply bring water to a boil and add it to the grain with occasional stirring and let the enzymes work.

Anyhoos, corn starch is something like 25% amylase and 75% amylopectin. Amylose molecules are more or less straight chain polysaccharides consisting of typically 200 to 20,000 glucose units. In the raw form amylose forms something of a helix shape as a result of the bond angles between the glucose units. The links between molecules are called 1,4-alpha glycosidic bonds. Whenever you bring grain to above the geletanization temperature, it is these straight chains that quickly swell up and form a collodial suspension turning everything into a nice gloppy gel.



Amylopectins are a branched form of amylose. It is different in that it has a branched nature due to both 1,6-alpha bonds and 1,4-alpha bonds.




Typically each branch is about 30 glucose molecules long with a branch about every 20-24 molecules long with a single amylopectin molecule comprising of up to 2 million linked glucose molecules!



Due to the tight bonding of the glucose molecules to each other, amylopectins are generally insoluble in water at room temperature. However, if the temperature is raised enough hydrogen bonding can begin to occur with water in essence causing the molecule to "poof up".

Alpha-amylase works by randomly cleaving the 1,4 linkages. Imagine chopping up a cooked spaghetti noodle with a knife by whacking away wildly at it... you end up with a bunch of smaller pieces of random lengths. This is great for amylose and this is how alpha-amylase works to liquefy the mash by busting up those colloidial suspensions of amylose. However, amylase can't chop the 1,6 bonds of amylopectins... it can only eat the end branches with 1,4 bonds up to the "the joint". If I am understanding correctly, this leaves so-called "limit dextrins".

The degree to which the starch has been hydrolized is called the dextrin equivalent or DE. Table sugar is glucose or dextrin and as such has a DE of 100. In the document below, researchers used concentrated liquid enzymes to hydrolize starch and evaluated the results based upon time and temperature.

PDF link

The highest yield was obtained at 300 minutes @70ºC using a .82 quarts to 1 pound grain ratio of all things. This is very thick and the reults were quite surprising... after 5 hours the mash had a DE of 28.4 with only 15% remaining starch. Whats really amazing is that 9% of the enzyme was still active after 5 hours at 70ºC.

As far as actual sugar conversion, glucoamylase attacks both 1,4 and 1,6 bonds and liberates one glucose molecule at a time like pac-man, but it is important that the starches be completely liquefied for maximum speed in and of conversion.

[HookLine]

Excellent info you're posting on this thread, DD.

(I also fixed the PDF link, wasn't working.)

[dixiedrifter]

So I decided to play with some numbers putting ethanol from plain old sugar vs corn in a cost comparison. Now my math skills prolly ain't too good with all the antihistamines and what not I'm taking so please bear with me.

Current references to industry yields indicate that most ethanol plants are on average yeilding about 18lbs of ethanol per 56lbs of corn. Sugar (assuming a real world isolation of 48% of 51% theory) would yield about 26-27lbs of ethanol per the same weight of corn.

So, for the industry average using corn one can assume it takes 3.1lbs feedstock to make 1lb of ethanol. Assuming a price of $6.50 per bushel and one achieving conversion to match that of the industry average it would cost about 36¢ to make one lb of ethanol, or bout $2.38 cents per gallon assuming there is 6.59lbs in a gallon of 100% ethanol.

For sugar it would take approximately 2.1lbs of sugar to make 1lb of ethanol. Based upon the cheapest price that I could find of 40¢ lb, it would cost about 84¢ or so to make one lb of ethanol, or $5.53USD per gallon using sugar as a feedstock.

But wait, there is more!

Lets say your using a turbo yeast to make your sugar wash and that each package is $5.25 after you pay for shipping. If you stretch the hell out of it, your looking at minimum of 3 packs to ferment 56lbs of sugar. So, suddenly your cost per lb goes up by about 61¢ or about $4.00 per gallon for a grand total of around $9.53 per gallon of 100%! OUCH.

Now then, looking at corn based ethanol and basing it on milehi's numbers it takes about 1 lb of alpha amylase to liquify 1000lbs of corn. Well for one, I don't trust that number unless I can see a certificate of analysis at the time of purchase and know the condition the enzymes were stored in. So to counter that, lets double that figure up to .9g of alpha amylase enzyme per lb of corn. Just for shits and giggles, lets also assume a reasonable even number price of $20.00 per lb of alpha-amylase after shipping costs. According to my calculations, thats about .044 cents per gram. Now then, it takes about 20.5lbs of corn to make a gallon of ethanol. So with the alpha amylase your looking at adding on another 82¢ to the price tag for a gallon of 100%.

Glucoamylase or Liquor Quik Whiskey Yeast with AG is about 1.9¢ per gram if you buy it in bulk after shipping, and IIRC the correct pitching rate of that is 3g per lb of grain. Add on $1.17 to the cost per gallon of 100%. This brings your grand total to $4.37 cents per gallon of 100% or roughly half the cost of sugar/turbo yeast. Due to the "idiot factor" if yeilds slip down 20% your looking at $5.34 per gallon. Still not cheap but a whole lot less than a turbo. I do believe however if you played with a pure sugar recipe some you could theroetically cut your costs down to just about that of corn maybe even lower.

But still corn has one major advantage over sugar no matter what the price... as you can't exactly go out and buy 200lbs of sugar without raising an eyebrow or two where as you can pick up a whole pallet of corn without so much as a blink.

[Dan Call]

Oh man.....you guys are going to laugh your a**es off at this, but I'm going to be honest. The first of the above mentioned mashes using just enzymes to convert. I got ONE PINT of drinkable beverage. The taste was very pleasing. It came off slowwwww too. Couldn't tune for a faster run, it was about a drop a second no matter what. I might have packed the collumn a little too tight but I purposefully lessened the packing. I feel like an idiot....it's sorta makes you wanna throw 50 pounds of Dixie Crystals in that fermenting batch to be sure....you know? But I"m resisting the urge.

I would like to point out that this is first time I've used Pint's temp control kit on the boiler....worked like a champ, very pleased with it. The fine grained adjustments are a little tough though when your temp is jumping all over the place. I finally wrapped several coats of tin foil around the collumn and it stabilized. I think the breeze blowing into the shed was effecting the collumn temp.

Of course...this could all be a likely story.

[Tater]

.85 qts water to lb grain seems like to little water to me .Would be hard to cook unless using a double boiler or steam.For hobby level stilling Id think more water be easier to deal with.

[Dnderhead]

I agree getting much over 1lb (1/2kg)to 2 qt (2l ) for most grains is hard enough . as with corn, wheat, oats and some others.
besides the grain will soak up much of that.
As soon as I git back to it (technical difficulties) I'm going to try 1Lb corn cooking that in 1 gal water then adding 1lb of malted barley cooking again
then adding another Lb malted barley to convert I thank that should give me about 8% wash. I use sparge water to cook with so I mite have to cut
back the next batch. aged in oak , I thank this should give me something like a irish whisky?

[kaziel]

Got me 25kg of corn, hope to make some decent liker with that. I'm not a big brew guy and don't want to use malt so I've go me enzymes as well. My local distillery sold me gluco and alpha. Alpha is a bacteria enz and cant stand temps up to 120 deg C. I've also received a "How to" paper. I should take a fine starch source in max 3:1 water/starch ratio, bring water to boil add half of alpha, slowly add starch bring it to boil, again after 10-15 minutes add second half of enz cook it on low fire for next 10-15 minutes. Then I should isolate it to retain heat after 4h when temp will drop to 60-65 deg C I should adjust PH to 4.2 I should add glucko stir it good and let it to cool for next few hours until below 30C, then add yeast.
I'm waiting to my PH strips to arrive, got citric acid to lower ph got me some calcium carbonate if I've will go to low. I will use 15L of water and 4kg of corn and se how that goes. I want to ferment "on the grains" and filter mash with nylon bag. I'm thinking about boiling corn for hour with alpha. Want to run the mash on my Boka with valve wide open it's a 1/8" so I've still got some reflux. I've got some questions and concerns hope you don't mind sharing your knowledge with me.
Q1. That this foams a lot during
a) fermentation
b) distillation
c) what can be used if so
Q2. Can this type of ashes can be used with immersion electric heated still (got 0.5% regulation up to 3.4kW)?
Q3. What's the best I can get out of that, I think about ABV (what's the yield like with methods described above)?

[Dan Call]

For some reason using just enzymes threatens your confidence in conversion. I have never iodine tested, but maybe I need to start.

My first "permit" mash is coming up and my attitude is complete different because I don't feel rushed or like I have to hide anything. I"m going to get this down to a replicable method to do it right every time, then branch out from there.

Here's my next move; I'm going to start adding an appropriate amount of 2 row barley into my corn mashes for diversity and extra conversion. I'm also going to add beano. In the first edition of Smiley's book he "required" beano then dropped it altogether in the second edition.

I have found that there are at least two different kind of alpha and gluco enzymes out there, one that can withstand higher temps and one that cannot. I think I have both kinds, but just ordered a fresh batch of the low temp kind. I've been told that enzymes will keep a long time, and you can just use a little more, and the recommended amounts are pretty small to begin with. I'm going to be looking for more clear syrupy liquid after conversion, as well as more sweet taste.

In mashes before I ever used enzymes, I would normally get a two inch layer of clear yellowy syrup on top, and it was very sweet. I don't know if enzymes are supposed to give this particular result. Between that and enzynes I'm expecting better conversion. I've also started sparging the corn, and I liked the taste better.

Anyone else using barley and enzymes in combination?

[Dnderhead]

I never had god luck with enzymes.they seem to lack something.
you also can try wheat base malt.

[aj2456]

HI all thought i would share what works for me on rice, using information from this thread:

First off i wouldnt recommend this as better than malt, definately do an Ag mash before this as this for me is harder, its just an option- as are yeast rice balls

i have the aplha and gluco from brewhaus/milehi i forget which one and a cheap temp probe and citric acid and ca chloride (fish tank grade)

i begin by cooking/ gelatinising the starches,

i then add calcium chloride- half a teaspoon seems about right for 3kg of rice and up to teaspoon of citric acid (start with less if in softer water and add more if not liquidating)) (or similar) this adds the ca ions for the enzymes and gets the Ph lower for the enzymes to kick in

speaking of which add the enzyme, ive not perfected the amounts but my pack says 0.1g to .3g per gallon , this seems like BS to me so i add about a 1/4 to 1/2tsp to my 3kg

if u have a bigass pot you can keep it all in there with the enzymes, i move mine into my mashtun and get it to 72 degrees c ideally and mix it all as best as i can (adding more water helps possibly with extra enzyme) and leave 20-40 mins/ start cooking another 3kg (my pot only holds 3kg)

if the alpha works it should noticably go mushy- sometimes v fast as you stir (this is a good day) if not even after being left, the temp was too high and it denatured, or the Ph was wrong- more acid can be added (tasting it helps, if noticably sharp dilute with water, add more enzyme and add some ca chloride)

after this steps good (if not sure/ had enough dont worry will still work but just not same efficency) lower the temp and Ph (more citric, to about 1tsp per gallon) (do want it noticably sharp now, think v dry harsh cider)

and when temp is 45degrees (can add cold water)

go ahead and add similar amounts of glucoamylase and leave (i leave mine overnight) till at pitching temps,

after about an hour with the glucoamylase it should have that nice clear syrupy layer (taste it, if not sweet something is wrong)

hints and tips
no need to grind rice,
lumps are bad, do best to kill at or during alpha amylase treatment
mix then mix some more
enzyme can be dissolved in warm not hot water before adding
dont add yeast till well cool else get suphoury badness

good luck