Lallzyme EX and EX-V

[crushday]

Quick question(s): Do either EX or EX-V have an inservice half life? Another way to ask, when it's added to the must when does it stop working? And, are there must conditions that lead to its expiration?

Are there "perfect" must conditions that accentuate extraction?

 

[Snafflebit]

I had the same questions. Enzymes being a complex protein will denature in high acidity, i.e. under 3 to 4 pH and deactivate above 80C (176F), in general. Lallzyme contains pectinase and cellulase. From the literature I have found, pectinase activity slows at 3pH and cellulase slows at 4pH.

These enzymes work quickly at 50C (or 120F) but stop working after ~2 hours at this temp. At about 30C, the activity rate drops by about half the 50C rate. At 18C pectinase will continue to work. But, time in the acid begins to reduce the lifetime of the enzyme also. I have read that an 8 hour cold soak of grape must and Lallzyme is enough, which I assume comes from experience because I cannot find any specific guidance from Lallemand.

As for expiration, I am seeing around 18 months if stored in a refrigerator and 6 months at room temperature and in powder form. If you had some older Lallzyme, you could assume that the half life is, I don't know, maybe 18 months if kept in the refrigerator and increase the dosage accordingly. I do not know of a problem with "overdosing" the must with enzyme. It is such a small addition anyway.

 

[Ajmassa]

I was under the impression that those enzymes are working for the duration of fermentation. Getting them in early (with or without cold soak) just gives them a head start to establish healthy reactions. And the more time on the skins the better—while mindful of not overdoing it for overly tannic wine adding years to aging. I’ve read accounts enzymes being added later during active AF to avoid tannin overload.

also I naturally assumed the effectiveness ebbs and flows with temperature changing, abv increasing, fruit condition, and just all the chemical reactions taking place during AF. Often at press time those skins will be mush- other times not so much. But to try and account for all these things is damn near impossible! So I just add in the recommended dosage and let the chips fall where they may. If fruit seems light I’d then try and stretch time on skins. If the skins look like mush then I’ll punch less vigorously.
Based off my limited experience it seems those enzymes will continue to work as long as there are skins present for them to break down

 

[crushday]

Here's my overriding observation that is driving the question. I used EX-V on my recent Cab Franc. I did not use EX-V on 20 gallons of Petit Verdot frozen must. I didn't think I would need to add it to the frozen PV must favoring the natural cell wall deterioration provided by freezing. The PV was started 7 days prior to the CF, however both were pressed on the same day.

I did not observe any noticeable difference in the condition of the skins from either the CF or the PV. I would think the CF would be broken down somewhat but that was not noticeable to me. It's a head scratcher for me, actually.

Does anyone else have experience in this regard?

 

[Johnd]

Here's my overriding observation that is driving the question. I used EX-V on my recent Cab Franc. I did not use EX-V on 20 gallons of Petit Verdot frozen must. I didn't think I would need to add it to the frozen PV must favoring the natural cell wall deterioration provided by freezing. The PV was started 7 days prior to the CF, however both were pressed on the same day.

I did not observe any noticeable difference in the condition of the skins from either the CF or the PV. I would think the CF would be broken down somewhat but that was not noticeable to me. It's a head scratcher for me, actually.

Does anyone else have experience in this regard?

Only anecdotal evidence for me. When I first used EX-V, it was on frozen musts that I was ordering and having delivered, and the enzymes went on board as soon as the must wast thawed, so in the mid 30's. My first use of the EX-V was an eye-opener, as I miscalculated my units somehow and used 10X the amount indicated in the instructions. My must was sludge at the end of fermentation and was a real PIA to press. I had to let the must sit in the basket nearly half an hour before it was solid enough to press without squishing out around the press plate. Lesson learned.

After that first error, I always got the dosage correct, and never had the problem again. When I switched over to fresh grapes, I continued to use the EX-V, and my grapes were always pretty chilly when I got them home and crushed them, the must was normally under 50 when it went in, and yeast was normally pitched the following day in the mid - 50's. Point being, I always ended up using it in pretty chilly musts, never put it into a warm must, so I can't verify the temp question. I've also never kept it longer than a season, always ordering what I needed for the upcoming season. Hope that helps at least a little...........

 

[Boatboy24]

First, thanks for brining this up, as it reminded me I forgot to order my Lallzyme for my upcoming crush. Second, I always thought the enzymes weren't working for too long, due to this comment in the MoreWine manual:

"Note: When using both Lallzyme-EX and fermentation tannins (VR Supra) in the must, we recommend adding the enzymes first then adding the tannins 6-8 hours after. This avoids the tannins from prematurely fining out the enzyme before it has a chance to work its magic on the grape skins."

I don't think one can assume that the enzymes will only work for 6-8 hours in the absence of fermentation tannin, but it would seem to me that most of the work would be done in that time, or the addition of tannin would be further delayed. This delay is also mentioned on the tannin page:

 

[crushday]

First, thanks for brining this up, as it reminded me I forgot to order my Lallzyme for my upcoming crush. Second, I always thought the enzymes weren't working for too long, due to this comment in the MoreWine manual:

"Note: When using both Lallzyme-EX and fermentation tannins (VR Supra) in the must, we recommend adding the enzymes first then adding the tannins 6-8 hours after. This avoids the tannins from prematurely fining out the enzyme before it has a chance to work its magic on the grape skins."

I don't think one can assume that the enzymes will only work for 6-8 hours in the absence of fermentation tannin, but it would seem to me that most of the work would be done in that time, or the addition of tannin would be further delayed. This delay is also mentioned on the tannin page:

View attachment 65820

Very interesting.

 

[Joe B.]

My first time using enzymes I did exactly what Johnd did and because of what the Morewine manual said about tannin's and enzymes I overloaded the must with oak power I had on hand and pressed a little early. I think the oak power helped with the mush issue some what.

 

[BI81]

The enzyme will complex with the substrate (pectin and cellulose in the cell walls) which allows the enzyme to act as a catalyst and increase the reaction rate to break down the substrate in the cell walls.

The enzymes are not changed by the reaction and are available to facilitate the reaction many times, but can only do so after the reaction is complete and the products have been released from the active site. In effect once the products are released the enzyme is "re-energizing" and able to accept another molecule and will continue to act as a catalyst as long as there is substrate available to do so.

The one variable that I don't know is how long the reaction takes and when the enzyme will be available to complex with another molecule and if this happens quick enough to allow for multiple reactions within the time frame of a fermentation.

 

[KCCam]

you have a simple chemical RXN of sorts going on between the Lallzyme (enzyme) and the grape skins (substrate).

which allows the enzyme to act as a catalyst and increase the reaction rate to break down the substrate in the cell walls.

So the question is:

Does the enzyme take place in the reaction, or is it simply a catalyst that speeds up a reaction but is not consumed by it.​

I am not a chemist, so I’m curious to know which one is correct.

 

[KCCam]

So the question is:

​

Does the enzyme take place in the reaction, or is it simply a catalyst that speeds up a reaction but is not consumed by it.​

I am not a chemist, so I’m curious to know which one is correct.

According to Enzymes and the active site (article) | Khan Academy

A substance that speeds up a chemical reaction—without being a reactant—is called a catalyst. The catalysts for biochemical reactions that happen in living organisms are called enzymes. Enzymes are usually proteins, though some ribonucleic acid (RNA) molecules act as enzymes too.​

I’m leaning toward it being a catalyst, as the link claims, because it explains why such a tiny amount can effect such a huge change.

I’m also quite happy to admit I’m wrong if convinced otherwise.

 

[Snafflebit]

Enzymes are catalysts, but enzymes are complex proteins and can be broken down like any protein, unlike other catalysts such as activated carbon or fined platinum

 

[BI81]

According to Enzymes and the active site (article) | Khan Academy

A substance that speeds up a chemical reaction—without being a reactant—is called a catalyst. The catalysts for biochemical reactions that happen in living organisms are called enzymes. Enzymes are usually proteins, though some ribonucleic acid (RNA) molecules act as enzymes too.​

I’m leaning toward it being a catalyst, as the link claims, because it explains why such a tiny amount can effect such a huge change.

I’m also quite happy to admit I’m wrong if convinced otherwise.

You’re correct an enzyme is a catalyst and is not changed by the reaction. It provides an active site to complex with a specific molecule (some are more selective than others ie will complex with more than 1 molecule) and reduces the activation energy required to complete a reaction. When the reaction is complete the Ezyme-Product Catalyst releases the product and is available to complex with another molecule.

 

[BI81]

Enzymes are catalysts, but enzymes are complex proteins and can be broken down like any protein, unlike other catalysts such as activated carbon or fined platinum

This is also correct all proteins (enzymes included) can be denatured by increasing temp past the optimal range, changes in pH and physical agitation. However in the reactions involving pectinase and hemicellulase during fermentation the reaction itself doesn’t denature the enzyme, as far as I know