How Much Lactase is Needed to Break Down One Gram of Lactose in 30 Minutes?

[Ouabache]

Can someone help me find out how much lactase will breakdown a gram of lactose? I asked at our public library, with no success.
(Please cite reference: e.g text, Biochem course, personal experiment data...)

I am trying to make a calculation and created thread lactase query on the chemistry topic, but have not received any replies. Perhaps the readership there has expertise in other areas, outside of biochemistry.

Thanks for your thoughts and comments..

 

[misskitty]

I'd like to know the answer to your question as well. It might help me manage better. Just out of curiousity, how are you making your calculations?

~Kitty

 

[Ouabache]

So as not to bias the replies to my query, I'll send you my present calculations, by private message.

 

[misskitty]

Makes sense. Still, why the interest in how much it will take to break down one gram of lactose?

~Kitty

 

[quetzalcoatl9]

do a pubmed search for kinetic studies of lactase, I'm sure you'll find what you're looking for.

i think it's a safe bet that the lactase will break down all of the lactose, it is just a question of how long it will take and the dependence upon concentrations of lactase and lactose.

 

[iansmith]

Kinetics will also depend on the temperature.

 

[Ouabache]

Makes sense. Still, why the interest in how much it will take to break down one gram of lactose?

~Kitty

Knowing how much lactase is needed to break down a specific weight of lactose, provides information to treat liquid dairy with any source of the enzyme, not just Lactaid caplets. As you may know, if you predigest the lactose in liquid diary, those with lactose intolerance can eat them without any concern.

 

[Ouabache]

do a pubmed search for kinetic studies of lactase, I'm sure you'll find what you're looking for.

i think it's a safe bet that the lactase will break down all of the lactose, it is just a question of how long it will take and the dependence upon concentrations of lactase and lactose.

Thanks for suggestion. I'm not really concerned how long the reaction will take, more that I am using enough enzyme. I also realize enzymes have a shelf life. It would be nice to know at what rate this potency diminishes? I imagine environmental factors (temp, humidity, time...) are variables of that equation.

Besides its probable affect on shelf life, iansmith points out that temperature is a variable of reaction kinetics.

 

[misskitty]

do a pubmed search for kinetic studies of lactase, I'm sure you'll find what you're looking for.

i think it's a safe bet that the lactase will break down all of the lactose, it is just a question of how long it will take and the dependence upon concentrations of lactase and lactose.

Sometimes the lactose doesn't always work. It has happened to me even when I have taken enough. Sometimes the lactase enzyme doesn't always break down everything even if you haven't pushed yourself over LI limit.

Typically with the over the counter Lactaid it says if you are eating foods containing Lactose for more than 30 minutes you'll need to take more lactaid for it to continue working. From personal experience I can say that Lactaid works for between 30-45 minutes. Never longer than 45 minutes. Usually it works for 35 minutes, less if the food you are eating has more lactose in it.

~Kitty

 

[quetzalcoatl9]

Thanks for suggestion. I'm not really concerned how long the reaction will take, more that I am using enough enzyme. I also realize enzymes have a shelf life. It would be nice to know at what rate this potency diminishes? I imagine environmental factors (temp, humidity, time...) are variables of that equation.

theoretically, one molecule of enzyme will catalyze the breakdown of the entire gram of lactose (if given enough time). see what i mean?

open a book to the topic of enzyme kinetics. look at a lineweaver-burk plot for instance. i would imagine that lactase has been well studied already.

Besides its probable affect on shelf life, iansmith points out that temperature is a variable of reaction kinetics.

as is pressure...but we really don't care about that. if we consider constant temp and pressure then what is important are the concentrations.

 

[misskitty]

Well yes considering that pressure and temperature has an affect on the concentrations of the lactase enzyme.

One molecule would breakdown a gram of lactose if it was potent enough and had a long enough period of time and if the conditions for that to happen were good.

~Kitty

 

[Ouabache]

theoretically, one molecule of enzyme will catalyze the breakdown of the entire gram of lactose (if given enough time). see what i mean?

open a book to the topic of enzyme kinetics. look at a lineweaver-burk plot for instance. i would imagine that lactase has been well studied already.

Yes I am thinking lactase would be well studied too. You are shaking a few cobwebs out of my head, mentioning Lineweaver-Burk plots. I just looked at a http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/E/EnzymeKinetics.html and recognize the general form from bygone biochemistry courses. Looking at them now, I realize I didn't fully appreciate the underlying principles of the variables. (But I could generate some great plots on exams )

Another idea I was reminded of, is that enzymes are catalysts. (also recall that from high school biology) They lower the activation energy needed to convert a substrate by binding with the substrate, changing its conformation to enable further breakdown to occur, without being changed in the process.

So how does the conversion of lactose proceed in a milk emulsion?
An NIH site describes, "Adding a few drops of the enzyme will convert the lactose in milk or cream, making it more digestible for people with lactose intolerance."
At least in bacteria (and reasonable assumption, we share this pathway); in the metabolism of lactose, other enzymes are synthesized and used in conversion, such as lactose permease which actively transports the sugar molecules into the cell; and beta galactosidase which cleaves lactose into galactose and glucose. These simple sugars can enter subsequent metabolic pathways to generate energy, etc..

So if these metabolic steps occur in vivo; I wonder what exactly does lactose convert to, in enzyme treated milk? Perhaps the lactase is just binding with all the lactose present, so the converted lactose is just "lactase-bound-lactose". Then we would surely need more than one molecule of lactase

 

[quetzalcoatl9]

Well yes considering that pressure and temperature has an affect on the concentrations of the lactase enzyme.

no, they don't. while the volume of the solution could change (and yes that could technically change the concentration), the mols of enzyme will be the same.

for example, you could compress the solution and cool it and get exactly the same kinetics. what pressure and temperature affect are the kinetic motions of the molecules in solution (from a classical perspective).

temperature or pressure could possibly, argueably, increase or decrease the solubility of the protein or solute, thereby raising the 'virtual' concentration, but that should not be the case in such a study since the solubility should be as close to complete as possible through use of ionic strength buffers, detergents, etc.

 

[quetzalcoatl9]

I wonder what exactly does lactose convert to, in enzyme treated milk?

galactose and glucose i believe.

 

[Ouabache]

galactose and glucose i believe.

How? Doesn't that require Beta galactosidase ?
I don't believe that enzyme is found in milk.

 

[iansmith]

How? Doesn't that require Beta galactosidase ?
I don't believe that enzyme is found in milk.

There bacteria found in milk that have beta-galactosidase. However, these bacteria are in smaller number in pasteurized milk and grow very slowing at 4C.

 

[quetzalcoatl9]

How? Doesn't that require Beta galactosidase ?
I don't believe that enzyme is found in milk.

lactase and galactosidase are the same enzyme - different names.

 

[Ouabache]

There bacteria found in milk that have beta-galactosidase. However, these bacteria are in smaller number in pasteurized milk and grow very slowing at 4C.

You're right.. there are some resident bacteria left in pasteurized milk.. I bet some of them can synthesize beta galactosidase.

 

[Ouabache]

lactase and galactosidase are the same enzyme - different names.

Hmmmm I wasn't aware of that.. As I research this on the web, it appears there is more to it.

Lactase has two properties: β-galactosidase activity and glucosidase activity.http://ethesis.helsinki.fi/julkaisut/laa/biola/vk/peuhkuri/chap2.html
But that does clear up the mystery (confusion) between chemical names.

Thanks for bearing with me, as I try to understand how enzymes facilitate transformation of a molecule.
I believe my above statement is still correct... "Enzymes lower the activation energy needed to convert a substrate by binding with that substrate, changing its conformation to enable further breakdown"

So when lactase attaches to lactose, the latter's conformation (3-dimensional shape) changes and then what? Does lactose just spontaneously cleave into galactose and glucose or is there some other molecule that drives this spit? Can this occur effectively at normal refrigeration; 34 to 40°F (1 to 3°C)?

 

[quetzalcoatl9]

I believe my above statement is still correct... "Enzymes lower the activation energy needed to convert a substrate by binding with that substrate, changing its conformation to enable further breakdown"

yes

So when lactase attaches to lactose, the latter's conformation (3-dimensional shape) changes and then what? Does lactose just spontaneously cleave into galactose and glucose or is there some other molecule that drives this spit?

you can look up the mechanism of action for the enzyme. enzymology is fascinating chemistry. undoubtablely, lactase is coordinating a water molecule (the enzyme certainly contains some metal atom like Fe for this purpose) in close proximity of the polysaccharide bond so that it can be hydrolized, but this is just my guess.

Can this occur effectively at normal refrigeration; 34 to 40°F (1 to 3°C)?

absolutely. people doing protein chemistry go to great lengths to store their proteins at temperature as low as -80 (being frozen at -20 is not enough!) to prevent degradation due to protease activity. the question is: at what rate?

 

[jim mcnamara]

a note to MissK -

The big reason Lacataid is active for a short time is that your stomach is proteolysing the lactase enzyme with proteinase enzymes. Lactase in humans (ones with lactase in sufficient quantities) is produced in the small intestine. Most of the enzymes in Lactaid don't make it that far.

Adding the enzyme to food gives it time to work in the absence of it being digested itself. - unless the food is at very cold temperatures.

 

[Ouabache]

So under normal refrigeration (1 to 3°C), how much lactase is needed to convert 1 gm of lactose in reasonable amount of time (say 30 mins)?

Also on related note, how much lactase is there in 1 caplet of Lactaid, given 3000 FCC Units/caplet? (What is FCC Units to mass conversion ??)