Enzyme Supplement Formulation
Letter for Doctors & Patients July 2001
I have spent the last decade working with various enzymes ranging
from exotic one such as thermal stable DNA polymerases to the
more common such as bromelain.1-3 As one with many years of experience
with enzymes, I am often asked which enzymes and how much should
go into a digestive product. I can understand this because frequently,
it appears as if there is really no rhyme or reason as to what
is in the blend. I am going to walk us all through how I would
go about designing a general enzyme.
A wide variety of enzyme supplements are currently on the market.
They are designed to address an equally wide range of health
issue. Such health concerns include conditions such as lactose
maldigestion, general gastrointestinal distress, flatulence,
autism, cancer, weight management, etc. Each one of these areas
is often addressed by a particular enzyme supplement. For example,
the use of lactase (beta-galactosidase) for the digestion of
lactose in lactose maldigesters. Many times, the particular enzyme
that is required, such as lactase, is presented by itself. The
choice then becomes very easy as to which enzyme one wants. However,
often times, there are five, ten, and even more enzymes in a
particular product. How does one decide which is right for them?
The answer lies in a more important question. How does one's
favorite supplement company decide what to offer in an enzyme
product? Further, how do they determine how MUCH of each enzyme
goes into that blend? Those are the two deciding questions in
enzyme supplementation and will be address below.
Please keep in mind that this is just a general digestive aid.
It is not meant to completely replace the body's own ability
to carry out some digestion. Rather, it is designed to compliment
There are several issues that
need addressing when developing an enzyme digestive aid. Most
notably are the targeted ages (i.e.
adults would more likely require lactase than juveniles) and
geographical location (i.e. the U.S. consumes more beef and dairy
than Asia on a per capita basis and might require a more aggressive
protease formula). Without further definition, I have made some
assumptions; 1) The consumer is an adult; 2) that adult's
diet is represented, on average, by the examples I have provided.
The examples cover the range of lacto-ovo (consume egg and milk
products) vegetarians to meat eaters. Additionally, each of the
meals would appeal to different levels of health-consciousness.
These examples seem reasonable but I have included the URL, http://www.nal.usda.gov/fnic/
for the USDA web site incase you prefer to find other representatives
for the diet. I am basing my calculations on the ones I selected.
That being said, I have decided the best approach is to take
the obvious facts first and that is, that there should be a Protease,
a Amylase, and a Lipase.
The enzymes should be as stable as possible over the broadest
pH range so they will be functional in both the gastric and intestinal
environments. The fungal enzymes fit these requirements better
than the bacterial and animal enzymes. Today, fungal enzymes
are produced and sold after much
First, the simplest way to do
this is to look at a typical nutritional box. From the Nutrition
Facts Box on foods, 2000 Cal/day is used
as the "average diet". We can make an assumption that
the average diet is 30% calories from fat (67 grams), 30% protein
(150g) and 40% CHO (200g). If 5.2% of the protein is tyrosine
that would give you 7.8 grams or 43,000 micromoles of tyrosine
For an Acid Stable Protease with an activity of 2,000 SAPU /
one SAPU is the amount of enzyme, which will liberate one micromole
of tyrosine per minute. How this relates to conditions in the
stomach and to digesting insoluble protein (e.g. a beef steak)
in the diet versus soluble protein (casein) has not been correlated.
a biomedical text book (White, Handler and Smith) there were
six animal proteins with an average tyrosine content of 5.2%
(range from 0.8 to 12.6%. How this average relates to the average
protein of someone's diet can be highly variable but it should
be in the right order of magnitude.
Assuming you have 30 minutes (more
on that later) to digest the food, you would liberate 30 micromoles
of tryosine per SAPU
during 30 minutes of digestion (assuming the enzyme is completely
stable for that 30 minutes). For 43,000 micromoles (see above)
of tryosine you would need 1,433 SAPUs or 717 mg of the Acid
Stable Protease. If you assume that the protein intake is divided
among 3 meals per day that would be 239 mg of Protease per meal.
Next, what about carbohydrates (CHO)? It is given that one Sandstedt
Kneen Blish Unit (SKBU) will liberate 1.0 grams of limit-dextrin
substrate per hour. Based on the activity, it is reasonable to
say one SKBU will digest on gram of starch per hour. With 40
g of CHO in the diet, and given an activity of 100,000 SKBU /
g, 0.4 mg would be required to digest all the CHO present in
Finally, it is known from our work
that 1.3 g of a lipase at 50,000LU / g ( or a total of 65,000
U) will digest 300 grams
of Olive Oil in 1 hour at 37C. Again, if one assumes all the
fat is from the oil, then the 67 g would take 14,517U (65,000U
/300g = xU/ 67g) to digest to completion. This is the amount
in 181.5 mg of Lipase Concentrate at the activity mentioned.
That all being said, we have completely
neglected any contribution by endogenous enzymes to the digestive
process. Again, an assumption
was made that we needed to completely supplement the diet. As
a starting point, this then allows one to easily calculate different,
moreindividualized, diet plans.
Next, the more difficult way to do
this is to find some actual examples of meals, which seem typical.
I have taken these off
the USDA website and your more than welcome to find other variations,
which may more closely suit your requirements. Three meals seem
- The BANQUET EXTRA HELPING
- A Glass (cup) of Milk, reduced fat, 2%, protein fortified.
Total protein: (29.083g + 9.717g) 38.8 g
Total fat: (40.045g + 4.871g) 44.916 g
Total carbohydrate: (33.567g
+ 13.505g) 47.072g
Based on the activity of acid stable protease, the amount of
enzyme required to digest the protein in this meal would be:
0.052 x 38.8g = tyrosine present = 10,970?M requiring 365.52
SAPUs for digestion or 182.76mg of the AFP at 2,000 SAPU/g would
Certain assumptions are required here
as well. From our assay for lipase activity, the simplest way
to do this is to assume
all the fat comes from Olive Oil. Without this assumption, the
problem becomes unwieldy in complexity. Given due consideration,
I believe you will find this a reasonable assumption for the
sake of the calculations. Based on the activity of our fungal
lipase, the amount of enzyme required to digest the fat, in this
meal would be:
If 67g of oil requires 181.5 mg of Lipase Concentrate, then
44.916g requires 135.22 mg.
Next, assuming all the carbohydrate
needs to be digested by an enzyme comparable to the human amylase,
I would suggest using
fungal alpha amylase. Based on the activity of this enzyme, the
amount of enzyme required to digest the carbohydrate in this
meal would be: Again, one Sandstedt Kneen Blish Unit (SKBU) will
liberate 1.0 grams of limit-dextrin substrate per hour. Based
on the activity,
it is reasonable to say one SKBU will digest on gram of starch
per hour. With 100,000 SKBU / g, and assuming all 47.072g of
carbohydrate in the meal is starch, one would need 47.072 SKBU
to complete the digestion in an hour or 0.4007 mg per meal /
- HEALTHY CHOICE Chicken Teriyaki with
Rice Medley, Mixed Vegetables in Butter Sauce and Apple Cherry
Compote, frozen meal.
- Milk, reduced fat, fluid, 2% milkfat, protein fortified, with
added vitamin A
Total protein: (17.006g + 9.717g) 26.723g
Total fat: (5.616g + 4.871g) 10.487g
Total carbohydrates: (37.097g + 13.505g) 50.602g
AFP required = 26.723g x 0.052 = 1.390g tyrosine = 7,555uM of
tyrosine requiring 125.87 mg of AFP per meal.
Again, if 67 g requires
181.5mg then 10.487g requires 28.4mg ofLipase @ 80,000 LU /g.
With 100,000 SKBU
/ g, and assuming all 50.602g of carbohydrate in the meal is
starch, one would need 50.602 SKBU to complete
the digestion in an hour or 0.506 mg per meal / capsule.
-GREEN GIANT, HARVEST BURGER,
Original Flavor, All Vegetable Protein Patties, frozen.
- Milk, reduced fat, fluid, 2% milkfat, protein fortified, with
added vitamin A
- 1 container Dannon Sprinkl'ins yogurts, fruit, low fat.
Total protein: (18.00g + 9.717g + 5.069g) 32.786g
Total fat: (4.140g + 4.871g + 1.253g) 10.264g
Total carbohydrates: (7.020g + 13.505g + 22.098g) 42.623g
32.786g x 0.052 =
1.704 g tyrosine = 9,393 ?M tyrosine = 156.48 mg of AFP is required
If 67g required 181 mg of Lipase, then 10.264 g would require
With a starting material
of say 100,000 SKBU / g, and assuming all 42.623g of carbohydrate
in the meal is starch, one would
need 42.623 SKBU to complete the digestion in an hour or 0.4262
mg per meal.
As a general rule-of-thumb, one glass (12oz) of milk requires
3.3 mg of Lactase (100,000 SKBU/g) to digest 80% of the lactose
under conditions of assay. That translates to 330 LACU / glass
of milk or per dosage. Again, that would be the amount required
for digestion over a very short period of time so the number
can be adjusted downward if a more extended digestion period
Finally, I suggest an inclusion of Phytase. Briefly, phytase
digests phytic acid. Phytic acid (Inositol hexakisphosphate)
is present in plant fiber and due to all the phosphate groups,
has a net negative charge. The negative charge affords good binding
to cations of nutrients such as calcium, magnesium, iron, manganese,
etc. Additionally, digestion of phytic acid allows greater bio-availability
of the freed phosphorus groups. Typically, 500 U are used per
Kg of feed. Humans typically do not eat a Kg at a time (on average),
so I considerably less. One tenth that amount seems reasonable
for the average meal so 50 Units should be sufficient. Phytase
is a fantastic enzyme and anyone concerned about mineral absorption
/ loss should look for it in their enzyme products.
What about Papain and Bromelain?
Both are good digestive enzymes. They fall into the same family
of cysteine proteases with similar
substrate specificities and functions. There should be no problem
substituting one for the other. However, I suggest "hedging
your bet" and splitting the activity between them. Bromelain
has been demonstrated to be an effective anti-inflammatory as
well as digestive enzyme, so if you are forced to pick one over
the other, I would recommend Bromelain. If one grinds through
the calculations to find some sort of equivalency for Bromelain
and papain a ratio that has worked well in our laboratory is
230 BTU for bromelain are equal to 4,500,000 PU for papain. Typically,
we suggest around 20-25 U of bromelain and 250,000 PU of papain.
There is a little more bromelain because, again, I tend to slightly
favor it in the formulations.
In addition, I would recommend alpha galactosidase (AG) to take
care of the oligosaccharides such as raffinose, stachyose, and
melibiose, which are often responsible for complains of flatulence
or indigestion. A 1:3 ratio of alpha amylase : alpha galactosidase
seems prudent given the high prevalence of gas forming foods.
As they are both naturally occurring carbohydrases in many of
the organisms used to produce commercial enzymes, they are both
amenable to digestive blends.
Finally, I said I would talk more about digestion time. This
directly relates to the final formula. The actual dosage used
in the formula can be scaled back somewhat from the numbers I
used above. That is because in the examples I gave, I used very
short digestion times. Also, to keep the calculations from becoming
unwieldy, I assume all the digestion had to come from the supplemented
enzymes. Those are valid assumptions and will help the formulator,
but now we need to apply it to real life. Therefore, one can
decrease the levels of those enzymes several fold and still have
an efficacious and affordable product. A typical final formula
might look something like this:
ENZYME BLEND EXAMPLE
Acid Stable Protease 240 SAPU
Alpha Amylase 15 SKBU
Lipase 6111 LU
Lactase 139 LacU
Bromelain 20 BTU
Papain 210500 PU
Phytase 52.6 U
Alpha Galactosidase 53 U
Please note that I did not throw in "everything but the
kitchen sink," as some blends seem to. For instance, cellulase
is omitted. The jury is still out on this but for a general digestive
blend, for people without any major problem, I don't see any
reason to add it. However, it some instances it could greatly
assist in the release of nutrients and should be considered based
upon the application.
Enzyme supplementation can be a wonderful tool as a digestive
aid.3 With a little time and attention, formulas can be created
to address a wide range of specific health. As for what units
to look for, I have used some of my favorites based on assays
I am comfortable with. The units that are used really do not
matter so long as one is able to calculate how much to use based
on the enzyme assay. All units are defined by their assay and
which ones are used usually is only a reflection of what any
particular laboratory is doing at any given time. Some people
look for FCC units but the problem is that some of the FCC assays
are antiquated and based on old techniques and equipment. Again,
the units are just labels of activity based on an assay. So long
as you have some label of activity ( and not just the mass) then
you can always do a conversion.
While the enzyme unit issue may
appear incomprehensible, it easily understood with a little
background. Armed with this,
you will be able to determine when someone is trying to take
advantage of your innocence by trying to confuse you with enzyme
units. Let's start with an example of a protease, an enzyme
that digests protein. A protease is not a protease is not a protease.
What I mean by that is while yes, A,B, and C protease can cut
protein X, they will probably all cut at different sites. Additionally,
they might use different reactive clefts in the enzyme to do
so. Also, they will all function different even in the same assay.
That functionality is itself a function of several parameters
such as time, temperature, and pH, not to mention, the structure
of the enzyme itself. Similar proteases can have different structures.
Those nuances are why there are different assays for seemingly
similar enzymes. A distillation down to the least common denominator
(i.e., saying they are collectively proteases and therefore the
same) causes a loss of meaning. That is what makes it look incomprehensible
and that is where many enzyme companies try to cash in by confusing
you. Each of those assays, is designed to be able to determine
the maximal amount of activity that enzyme is capable of, under
those conditions. Those conditions, usually are dictated by food
applications, because the lion's share of enzymes sold in the
world is for food processing. Though, detergents may be close
but that we are really not interested in that. It is not a good
idea to base enzyme assays on how much blood detergent-X will
get out. That would *really* flip consumers out. As an aside,
what makes colors bright, is cellulase. It digests the textile
frayed strands and makes them look sharper. Anyway, that issue
is being addressed in that way.
You see, in enzymology, for all
the reasons listed above, each lab has to determine how the
enzymes perform for them. The units
listed on a specification sheet only give an idea of where to
start. When one gets enzymes "in" from a company, what
is usual is to take that enzyme, read the data sheet and set
up a standard curve of your own. Slight variations in methods
(and that is common from lab to lab) can have dramatic differences
in how the enzymes perform. Your lab may be at 23C and mine at
25C. That *will* make a notable difference in some cases. Those
can be minimized but not eliminated by the scientific method.
Also, the application is vital. That standard curve just gives
the lab an idea of the activity that enzyme will have in their
lab. Then, the lab/company needs to determine how well the enzyme
performs in their application. Again, they then, usually, do
serial dilutions and assay the enzyme in the application. Often
times, they make up their own units during these assay procedures
so that they mean more to them. For instance, who cares how many
tyrosines a protease will liberate in one minute if you are trying
to debitter a powdered drink mix, for instance. Answer: no one.
It might give an approximation on where to start with your activity,
but it really means very little. What will usually be done, is
that lab will create a unit definition in their books based on
debittering of protein. In a theoretical case, the assay is really
how well the blend works in the application of being a digestive
aid in humans. That being said, one suggests starting with a
low concentration of the blend (half a cap or a quarter cap...whatever)
and then working up until the desired effect is observed. That
is how to establish a 'standard curve' if you will. That is the
way good enzymology is done. Period.
Recognizing this, the Enzyme
Technical Association (ETA), the decided governing body on
enzymes in the United States states on their website, under Industry
Guidelines for the Use of Enzymes in Dietary Supplements:
VI. ENZYME ACTIVITY METHODS FOR DIETARY SUPPLEMENTS
In order for consumers and health professionals to be able to
compare enzymes, both in the commercial market place and in clinical
and other published studies, it is necessary that enzyme potencies
be expressed in scientifically sound units of activity.
The ETA guidelines for enzyme activities
1. Enzymes in dietary supplements should be measured and
labeled on an activity basis, not a weight basis.
2. An authorized, compendial method of measuring and expressing
enzyme activity such as Food Chemicals Codex (FCC), United States
Pharmacopoeia (USP), Federation Internationale Pharmaceutique
(FIP) or Japan Pharmacopoeia (JP) should be adopted whenever
3. For enzymes not adequately covered in compendial sources,
it is recommended that activity methods that are used have undergone
scientifically sound development and validation procedures.
4. When developing new assay methods, widely available equipment
and reagents should be used.
5. The assay temperature should be body temperature, 37oC, unless
characteristics of the enzyme preclude this temperature.
6. An assay pH range of pH 4 to pH 5 is recommended whenever
possible if the enzyme is to be ingested.
7. A well-defined substrate with adequate lot-to-lot uniformity
should be used.
8. At a minimum, validation of enzyme assays should document
assay specificity, assay variability, assay linearity and assay
Paying special attention to numbers 2 and 3, one can see that
there are some recommended federations from which to use enzyme
assays and there is also recognition that not all areas are covered
by those. For enzyme units that are not included in the compendial
sources mentioned in number two, then number three should apply.
The qualification being that those assays are properly validated,
which is exactly what any good enzyme laboratory will do anyway.
I have sketched some broad strokes and covered a lot of ground
for you. Both patients and doctors should be asking the questions
I raised to make sure their enzyme product is really formulated
in their best interest. I have provided enzymes with units of
activity that I prefer because they are common in the food industry.
Others may, of course, be used but we have found in our laboratory
that those presented work best.
1. Miller, K.S. and Brudnak, M. (1994) Expression Cloning: PCR
Versus Episomal Vectors for Rescue of Transfected Genes. In
PCR in Neuroscience (Methods in Neuroscience Vol. 26) Volume
Editor G. Sarkar, Academic Press, Orlando, FL.
2. Mark Brudnak 2000.
Enzyme Therapy - Part I Townsend
Letter for Doctors & Patients.
3. Mark Brudnak 2001 Enzyme Therapy
- Part II Townsend
Letter for Doctors & Patients. January
Please feel free to contact
Mark A. Brudnak PhD, ND
957 Lake Shore Road
Grafton, WI 53024
(Wisconsin is in the Central