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junglejimmy

Using amylase to replace malted grains

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Hello

I was wanting to replace my malted wheat with enzymes for cost reasons. It is my understanding that the malted wheat in vodka production is used solely for its enzymes, so replacing it shouldn't have a huge effect on taste.

From my research I have found i need an alpha amylase for liquification (breaking starch down to dextrose), and a beta amylase (breaking dextrose down to glucose).

This brings me to my first question. There are three options available for alpha amylase: high, mid and low temperature alpha amylase. My guess is that the mid tempurature is most suitable as the range is closer to gleatinization temp, you want it to start working before the beta amylase and you don't want to waste time and energy heating the wort to excessive temps.

Also there are two options for the form of the enzymes. One is in powder form and the other is a brown liquid that looks like a syrup. Is one more suitable than the other in relation to quality and taste? Or is it merely a matter of how you like to store your enzymes?

Thanks

Jimmy

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I'm most familiar with the Specialty Enzymes liquids. No reason not to use the high-temp SEBstar HTL alpha as its optimal temp range is 50C to 90C. If you ever experiment with corn, you'll need it. One thing about using enzymes is they can be crazy particular about pH. Have a good meter and be prepared to make adjustments between alpha and gluco.

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Thanks 3dog,

I've decided to go with the high temperature resistant alpha-amylase and a soybean beta-amylase.

The optimal Ph range for the alpha is 6-6.5, which is rather narrow. I should be able to stay within it with some citric acid and a pH meter. The optimal temp is 90C in the specs. Do you think I will still have a good conversion at the gelatinization temp of 74C?

The soybean beta-amylase has an optimal Ph of 5-7.2. That's well with the range of the alpha. Hopefully I wont have to make adjustments. The optimal temp is between 50 - 60, so I should be right to add it after the alpha rest phase.

I do have one question however, does temperature have a large effect on Ph meter reading. if I have to adjust the Ph after the alpha at around 60C?

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I use a Hanna pHep 5 - it couldn't be any easier to take temperature adjusted pH readings, since it does it automatically. Just be sure to calibrate it every week or two (use 4.01 and 7.01, since that's the range we work in).

Only takes a second, adjusting with lactic or citric only takes a few minutes. Add small amounts and test until you get a good feeling for exactly how much acid you need to add. Keep the agitator running, and wait a few minutes between additions and testing to be sure you don't overshoot. It honestly doesn't make any sense to try to manage complex calculations to find out exactly how many ml of what molar acid you need. Trial and error until you get a handle. I don't know your batch sizes, but I will stress, if using trial and error, a little goes a long way.

It's relatively easy to adjust pH during the mashing process to hit the optimal enzyme pH with each step. Optimal temp and pH is different across Alpha Amylase, Beta Amylase, Glucoamylase, and Beta Glucanase - those might all be different from the pH you want at pitching time. Depending on what you are doing, you might find that multi-step acidification is useful.

Print out the spec sheets for all the enzymes you are planning to use, study them. They'll all have the optimal pH and temp, as well as the active range and denature temp/pH.

I'd say go with the HTAA, it's more versatile, skip the Beta, replace it with Glucoamylase, since conversion will be better. Unmalted wheat you might find you need to add Beta Glucanase to keep the mash viscosity manageable.

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Read the specs on your enzyme, but in my experience, pH is more important than temp. These buggers work FAST. You can literally watch the conversion happen within seconds of dumping high-temp alpha into a pot of corn goo.

Temperature makes a HUGE difference on pH. The Hanna pHep 5 jamesbednar mentions is a great bang for the buck, but do note that any meter must be used within its temp compensation range. the pHep 5 has an upper limit of 60C for readings.

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Thanks James,

I did not know the Ph and temp meter existed. That makes life so much easier.

We have 1000L fermentation tanks at our distillery. After doing a not of research and listening to you guys I think I Jane a plan of attack.

One other thing though James, you mentioned glucoamylase and beta-gluconase. I have read so much conflicting and confusing info about enzymes. From what I gather glucoamylase performs the same job as beta-amylase. I am unfimiliar with beta-glucanase, however after a bit of reaserch I came up with this:

"Beta-glucanse is part of the cellulose enzyme family, and are used to break up the beta glucans in (un)malted wheat, rye, oatmeal and unmalted barley."

Do you think I should add a beta-glucanase if I am going to use an all wheat vodka? And if so how would it be implimented?

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Glucoamylase is preferred over beta-amylase because it works on a wider range of starch bonds. It'll break 1-4 and 1-6, whereas beta-amylase work only on 1-4, leaving some starches behind as unfermentable.

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Also note the actual pH of a solution varies with temperature. This is NOT related to the ATC of the probe, which compensates for how the probe itself electrically reacts to temperature. Calibration of a pH meter should be done at the same temp as your usual readings. So, if you normally take readings at 50C, your calibration solutions should be heated to 50C before use.

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Thanks bluestone. I read somewhere that the increased maltose concentration from beta-amylase effects flavor in a good way.I think that is for beer though and probably wont effect the taste when doing a reflux run for vodka.

Also heating the calibration solutuons to high temps seems a little tricky if you want to reuse them. I know they usually come in little plastic containers. Maybe I could put them in a microwave for a minute. How do you heat them up?

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No problem. I missed the question about beta-glucanase. The enzyme company will provide you with instructions for everything.

Beer guys have to consider mouthfeel and stuff but I don't know much about that.

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I use the powdered calibration packets and mix my own (with distilled water) in mini mason jars.

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I believe White Labs has an enzyme that will yield close to 100% of the available sugars within barley; excluding fiber and cellulose (obviously).

The standard process in large ethanol distilleries involves a backwards mashing process, beginning with conversion using alpha-amylase, which makes unfermentable sugars, then moving down to glucoamlyase which is pitched with yeast and works actively as fermentation continues.

The ideal yields are measured in absolute alcohol per ton of grain.

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So we have everything pretty much laid out for you already OP but I'll rehash a little bit and consolidate.

Typically you are looking at a 3 step conversion process to turn grain starch into fermentable sugars.

  1. Gelatinization – Process of solubilizing starch granules in water. Typically accomplished by grinding grain and heating in the presence of water.
  2. Liquefaction – Initial breakdown of solubilized starch. Converts starch into dextrins (random sugars)
  3. Saccharification – Final breakdown of dextrins into fermentable sugars.

These have to be done in order or, in some cases, simultaneously. It is not a good idea to try to saccharify liquefied starch, nor is it a good idea to try to liquefy un-gelatinized starch.

Enzymes do not assist gelatinization typically. They are generally used for liquefaction and/or saccharification.

Liquefaction we are talking alpha-amylases. Of which there are thee main temperature ranges (already listed in above posts). The ideal part about using a high temperature alpha-amylase is that simultaneous gelatinization and liquefaction can take place at 80-90 C.

Saccharification we are talking beta-amylases or glucoamylases (every enzyme that has “amylase” in it will work on starch because starch = “amylose”). Beta-amylases are common in brewing (as it is found in malted barley) and will work to produce maltose, glucose, and other unfermentable sugars. Glucoamylases are frequently used by distilleries because it will convert all dextrins (random sugars) into glucose. Additionally glucoamylase has a side 1, 6 activity which will allow further degradation of some sugars that were previously unfermentable.

Beta-Glucanases on the other hand are a hemicellulase that will work to break down a very specific compound found commonly in wheat, barley, rye, and oats called beta-glucan. Beta-glucans can cause viscosity issues and gum up a mash or an immersion heater. This enzyme has little to no effect on starch and sugar conversions/yields.

TL: DR. Alpha-amylase is used for Liquefaction, Glucoamylase is good for Saccharification, and Beta-Glucanase is generally only applicable to rye, wheat, or barley mashes.

Please consult your enzyme provider for specific pH and temperature ranges of individual enzymes.

Cheers!

CDE

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I forgot to mention.

For a 100% raw wheat mash, you would probably want to look into a medium temp alpha-amylase, glucoamylase, and beta-glucanase.

High temp alpha-amylase would work as well, the higher temps might not be worth their cost considering the lower gelatinization temp of wheat.

That being said, if you plan on mashing other grains on a regular basis (like corn) it might be easier to have one procedure rather than several...

CDE

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Thanks CDE,

That is an amazing overview. You should write a book!

I noticed one thing you said about grinding however. Previously I have been steaming and rolling my wheat before mashing. I have a grinder, but even on the lowest setting the wheat comes out almost like flour.

Which method do you think is more suitable for an all grain vodka using enzymes? steaming and rolling or grinding?

Thanks.

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There is nothing wrong with having the wheat as a flour as long as you don't end up with lumps in your mash. You will get a better conversion and a faster one with a fine grind. Enzymes are the way to go, but with wheat mashes you will need to make sure your nutrients are adaquate. You may need to add some zinc and di ammonium phoshate isn't a bad idea either. It takes very little and it helps with yield. Sprouted or malted wheat will save some cost of enzymes, but that alpha amalysis is much more heat sensitive. Don't try to push for to high of an alcohol content 10%-11% max and a slower fermentation gives you a better spirit with less congeners to deal with. We use enzymes from Novozymes and their enzymes work very well but as stated before pH is important for best results.

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Hey Jimmy,

Sorry for the delay in getting back to you, maybe I will write a book one of these days…

What “miller” said is correct, grinding to a flour provides higher extract on average, and is generally the best way to fully utilize the starch on simpler systems. However flour is a bit harder to work with (clumping and fire hazard) compared to cracked or rolled grain. Steaming and rolling is also a good method of getting your grain cell walls disturbed enough to extract the starch, though perhaps not as thorough as grinding.

At the end of the say the take home is as such: The best method to use on your system, is the method that works best on your system.

Though there might be better ways to theoretically get higher yields, it may not be possible on your system.

Though “miller” is correct that sprouted of malted grain would save on cost of enzymes, it is important to point out that sprouted or malted grain is usually much more expensive than raw grain, so you would actually spend money avoiding enzymes. Raw grain conversions with enzymes is the cheapest and arguably the most effective method for starch conversion currently (go ask an ethanol plant).

Cheers!

CDE

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CDEs comments are indeed good, but let me add a twist wrt gelatinization.

First point - I am not addressing the issue of 'gums' mostly polymers of pentosan sugars that cause a lot of viscosity in the case of rye, for example.

There are two categories of grain starch, amylose and amylopectin. Amylose is mostly 'straight' chain glucose with 1,4 bonds and typically ~300 glucose molecules in length. Amylose is soluble in water and doesn't cause anything like the viscosity problems of amylopectins. I write 'straight' however they are really a spiral (like DNA) and the spiral traps iodine for the iodine starch test.

Amylopectin is not a real pectin (does not contain regular terminal methyl groups like fruit pectins), but a single grain starch granule may consist of ~300,000 glucose (up to 1.2mill units) units in a tree structure. The tree consists of segment of amylose (1,4 bonded glucose) typically around 18-22 glucose in length, which then split in a 'Y' with a 1,4 and a 1,6 bonded group. [imagine a 14-15 stage binary tree w/ 20-unit amylose units per segment] .The amylopectic is the stuff that absorbs the water in gelatinization. As the tree unfolds, the very polar water molecules become trapped between the branches of the 'Y' (the glucose hydroxyl groups are the cause). Amylopectin is the stuff that thickens a gravy or a mash as the starch granules swell, expand and trap water.

Common grains consist of ~70-80% starch (maize is on the top end, most small grains closer to 70%) and the starch is typically ~25% amylose and ~75% amylopectin. The exception is that "waxy" grains and especially waxy maize can be ~100% amylopectin.

So as you cook starch in a water slurry the amylopectin starch begins to unravel and trap water. If there is insufficient water then retrogradation sets in which the internal tangling of the amylopectins. A common example of retrograded starch is stale bread. Another is the 'skin' that forms on wet dough if allowed to dry. The retrograded starch is not susceptible to enzyme or acidic hydrolysis - it's indigestible and a distillers loss.

So the practical deal is that w/o enzymes, grain grist may require 10x to 12x the grist mass of water to prevent retrogradation during cooking. That means to prevent retro' you can only add ~2/3rd lb of grist to a gallon of water! The solution to get to ~2lb/gallon is to add a little 'debranching' enzyme to the grain cooker and this can drastically reduce the amount of water needed, as the debranching enzymes cut apart the amylopectin tree and reduce the water trapped. Alpha-amylase is a choice. I can't speak to the proprietary 'visco' enzymes, but they are certainly the right direction to create normal cereal cooking.

Enzymes do not assist gelatinization typically. They are generally used for liquefaction and/or saccharification.

Enzymes don't directly cause or improve gelatinization (that's a matter of water, heat and pH) but enzymes are necessary for a practical thick cereal cook w/o excessive retrogradation.

We should also mention beta-glucanases which assist in clearing the cellulosic "wrapper" from around the starch granule mega-molecules. This is fairly effective in malting, but for raw grists a rest may have some advantage. The starch will gelatinize at temperature in any case, but the glucanas residues can create a gel and add to viscosity.

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