Bacterial Protease

Description

HT-Proteolytic is a food grade proteolytic enzyme (E.C.3.4.24.4) derived from Bacillus subtillis var. HT-Proteolytic is a highly specific endopeptidase which is active on the interior peptide bonds of gluten proteins. The enzyme complies with the FCC and WHO/FAO (JECFA) recommended specifications for food grade enzymes. Unstandardized HT-Proteolytic is useful in tableting, blending, and other applications where high activity and low bulk are desirable.

HT-Proteolytic is an unstandardized concentrate which is sold on an activity basis. The activity varies over the range 2500 - 4500 NU/g. HT-Proteolytic 200N is standardized at 200 NU/g and 937,500 MWU/g of amylase.

Properties

	HT-Proteolytic Conc.	HT-Proteolytic 20ON
Form	Amorphous Dry Powder	Amorphous Dry Powder
Color	Light Tan	Light Tan
Odor	Free of Offensive Odor	Free of Offensive Odor
Taste	Free of Offensive Taste	Free of Offensive Taste
Solubility	Readily Water Soluble	Readily Water Soluble
Activity	2500 - 4500	200 NU/g, 937,500 MWU/g

Activity

One Northrop Unit (NU) is that activity which gives 40% hydrolysis of one liter of casein substrate in 60 minutes under the conditions of the assay.

A copy of the Colorimetric Northrop Assays available upon request.

Inhibitors

The protease in HT-Proteolytic is a metalloenzyme having zinc as a cofactor. It is inhibited by EDTA, cisteine, phosphates and urea. Heavy metal ions are also inhibitory.

Effect of pH

The protease component in HT-Proteolytic exhibits maximum activity at pH 6.5 - 8 at 40C and exhibits good activity over the pH range of 5.5 to 9.0. The enzyme is stable in dilute solutions over the range of pH 5.5 to 1 0.0 at temperatures below 50C. At temperatures above 60C, the enzyme is rapidly inactivated.

Optimum pH range	pH 6.5 to 8.0
Effective pH range	pH 5.5 to 9.0
pH stability	pH 5.5 to 10.0

Effect of Temperature

In the presence of substrate, HT-Proteolytic exhibits maximum activity at 45 - 55C and is stable in dilute solutions at temperatures up to 60C.

Maximum activity	45 - 55C
Stability	up to 60C

Inactivation

The protease component in HT-Proteolytic may be inactivated by raising the temperature to 80 - 85C and holding for 5 to 10 minutes. If high temperature inactivation is undesirable, the enzyme may be inactivated by adjusting the pH below 5.5 or above 10.0 and holding for 30 minutes at 60C.

Applications

Baking	To improve the mechanical properties of dough, especially for cookies and crackers
Feed Supplement	To improve animal feed utilization and efficiencies
Leather	Dehairing and bating of hides
Photographic	Removal of gelatin from film and silk screens

Use Level

Enzyme requirements are generally dictated by processing conditions. They will vary with protein substrate concentration, degree of desired protein hydrolysis, pH, temperature, and time. In general, increasing the HT-Proteolytic bacterial protease concentration will increase the rate of protein hydrolysis and shorten the time necessary for hydrolysis.

Soda Crackers

While the exact use level of HT-Proteolytic in baking varies with the type and protein content of the flour, the desired degree of modification, and the variety of baked good, we recommend initial evaluation be conducted at the following levels:

In the sponge and dough method, evaluate 1600 - 3200 NU per 1000 kilogram of flour; 8-16 gram HT-Proteolytic 200 or 0.5 - 1.0 gram HT-Proteolytic Concentrate (~3200 NU/g) per 1000 kilograms of flour.

In the dough method, evaluate HT-Proteolytic at 20,000 to 40,000 NU per 1000 kilograms of flour; 100-200 gram HT-Proteolytic 200 or 6 - 13 grams HT-Proteolytic Concentrate (~3200 NU/g) per 1000 kilograms of flour.

Storage Stability

In sealed containers, under cool dry conditions, the loss in activity of HT-Proteolytic is normally less than 10% over one year. Storage life can be extended by storing under refrigeration at 50C.