US4299848A — Modified enzyme system to inhibit bread firming m…

Claim 1

Independent Claim 74 words

A method comprising the steps of: providing a solution of alpha-amylase enzyme and protease enzyme; adjusting the solution with about 0.0006 M calcium sulphate and about 0.8 M sodium chloride to a pH of about 6.5; heating the solution to about 75° C.; adding to the solution from 1-3% wheat starch and the mixture adjusted to a pH of about 6.5; and holding the solution from 15-30 minutes to inactivate the protease enzyme activity.

Claim 2

Independent Claim 74 words

A method comprising the steps of: providing a solution of alpha-amylase enzyme and protease enzyme; adjusting the solution with about 0.0006 M calcium sulphate and about 0.8 M sodium chloride to a pH of about 6.5; heating the solution to about 75° C.; adding to the solution from 1-3% wheat starch and the mixture adjusted to a pH of about 6.5; and holding the solution from 15-30 minutes to inactivate the protease enzyme activity.

Claim 3

Independent Claim 31 words

A method comprising the steps of: providing an alpha-amylase enzyme free of protease activity; and adding the alpha-amylase to chlorinated tap water in the presence of at least 2.0-5.0% wheat flour.

Claim 4

Independent Claim 31 words

A method comprising the steps of: providing an alpha-amylase enzyme free of protease activity; and adding the alpha-amylase to chlorinated tap water in the presence of at least 2.0-5.0% wheat flour.

Claim 5

Dependent Claim 14 words

The method of claim 2 wherein the water has a pH of from 3.50-6.10.

Claim 6

Independent Claim 50 words

A method for the introduction of alpha-amylase to chlorinated tap water comprising the step of: adding the alpha-amylase which is free of protease activity to the tap water in the presence of at least about 0.3 mg. of ferrous sulfate per 100 ml. water at a pH above about 6.50.

Claim 7

Independent Claim 50 words

A method for the introduction of alpha-amylase to chlorinated tap water comprising the step of: adding the alpha-amylase which is free of protease activity to the tap water in the presence of at least about 0.3 mg. of ferrous sulfate per 100 ml. water at a pH above about 6.50.

Claim 8

Independent Claim 40 words

A method of introducing a dry enzyme concentrate to wheat flour comprising the steps of: suspending wheat starch in acetone, adding protease-free alpha-amylase to the acetone, separating the starch from the acetone, and blending the separated starch with wheat flour.

Claim 9

Independent Claim 40 words

A method of introducing a dry enzyme concentrate to wheat flour comprising the steps of: suspending wheat starch in acetone, adding protease-free alpha-amylase to the acetone, separating the starch from the acetone, and blending the separated starch with wheat flour.

Claim 10

Dependent Claim 18 words

The method of claim 5 wherein there is provided about 3405 DU alpha-amylase per 100 lbs. of flour.

Claim 11

Independent Claim 60 words

A method of inactivation of protease enzyme in a mixture comprising alpha-amylase enzyme and glycerol and water comprising the steps of: adjusting the mixture to a pH from 7.50-9.50; adjusting the ratios of glycerol to water from 6.0-1.0:1 by weight; and heating the mixture to a temperature of from 92°-95° C. for a time sufficient to inactivate the protease enzyme.

Claim 12

Independent Claim 60 words

A method of inactivation of protease enzyme in a mixture comprising alpha-amylase enzyme and glycerol and water comprising the steps of: adjusting the mixture to a pH from 7.50-9.50; adjusting the ratios of glycerol to water from 6.0-1.0:1 by weight; and heating the mixture to a temperature of from 92°-95° C. for a time sufficient to inactivate the protease enzyme.

Claim 13

Dependent Claim 13 words

The method of claim 7 wherein the mixture is held from 15-30 minutes.

Claim 14

Dependent Claim 10 words

The mixture of claim 7 wherever the ratio is 2.33:1.

Claim 15

Dependent Claim 68 words

In a mixture comprising alpha-amylase enzyme, protease enzyme, glycerol and water, a method of inactivation of the protease enzyme activity while retaining the alpha-amylase activity comprising the steps of: adjusting the mixture to a pH from 7.50-9.50 and a glycerol to water ratio of from 6.0-1.0:1 by weight, heating the mixture from 92°-95° C., and holding the heated mixture from 15-30 minutes until the protease enzyme is inactivated.

Claim 16

Dependent Claim 68 words

In a mixture comprising alpha-amylase enzyme, protease enzyme, glycerol and water, a method of inactivation of the protease enzyme activity while retaining the alpha-amylase activity comprising the steps of: adjusting the mixture to a pH from 7.50-9.50 and a glycerol to water ratio of from 6.0-1.0:1 by weight, heating the mixture from 92°-95° C., and holding the heated mixture from 15-30 minutes until the protease enzyme is inactivated.

Claim 17

Dependent Claim 10 words

The method of claim 10 wherein the ratio is 2.33:1.

Claim 18

Dependent Claim 59 words

In a mixture of alpha-amylase enzyme and protease enzyme and glycerol and water, a method of inactivation of the protease enzyme comprising the step of: heating the mixture to a temperature of from 92°-95° C. at a pH from 7.50-9.50 and at a glycerol to water ratio of from 6.0-1.0:1 for a time sufficient to inactivate the protease enzyme.

Claim 19

Dependent Claim 59 words

In a mixture of alpha-amylase enzyme and protease enzyme and glycerol and water, a method of inactivation of the protease enzyme comprising the step of: heating the mixture to a temperature of from 92°-95° C. at a pH from 7.50-9.50 and at a glycerol to water ratio of from 6.0-1.0:1 for a time sufficient to inactivate the protease enzyme.

Claim 20

Independent Claim 56 words

A method of introducing a dry enzyme concentrate to wheat flour comprising the steps of: suspending wheat starch in acetone, adding protease-free alpha-amylase to the acetone, separating the starch from the acetone, and blending the separated starch with wheat flour, wherein the protease-free alpha-amylase is made according to the method of claims 7, 10 or 12.

Claim 21

Independent Claim 18 words

The method of claims 7, 10 or 12 wherein the alpha-amylase and protease is provided as a powder.

Claim 22

Independent Claim 23 words

The method of claims 7, 10 or 12 wherein the alpha-amylase is selected from the group consisting of Bacillus subtilis and Bacillus sterothermophilis.

Claim 23

Independent Claim 42 words

The method of claims 7, 10 or 12 after inactivation of the protease enzyme, further including the steps of: adding the alpha-amylase to a NaCl solution, and fractionating the alpha-amylase by the addition of alcohol thereby precipitating a fraction of the alpha-amylase.

Claim 24

Dependent Claim 14 words

The method of claim 16 wherein the NaCl solution is about a 5% solution.

Claim 25

Dependent Claim 13 words

The method of claim 16 wherein the alcohol is a short chain alcohol.

Claim 26

Dependent Claim 18 words

The method of claim 18 wherein the alcohol is selected from the group consisting of methanol and ethanol.

Claim 27

Dependent Claim 21 words

The method of claim 16 wherein the alpha-amylase fraction is that precipitated by increasing the alcohol content from 50% to 66.7%.

Claim 28

Dependent Claim 11 words

The method of claim 20 wherein the content is from 50-64.3%.

Claim 29

Dependent Claim 11 words

The method of claim 20 wherein the content is from 50-61.5%.

Claim 30

Dependent Claim 11 words

The method of claim 20 wherein the content is from 50-58.3%.

Claim 31

Dependent Claim 11 words

The method of claim 20 wherein the content is from 50-54.5%.

Claim 32

Dependent Claim 11 words

The method of claim 20 wherein the content is from 54.5-61.5%.

Claim 33

Dependent Claim 11 words

The method of claim 20 wherein the content is from 54.5-58.3%.

Claim 34

Dependent Claim 11 words

The method of claim 20 wherein the content is from 58.3-61.5%.

Claim 35

Dependent Claim 12 words

The method of claim 16 wherein the NaCl solution contains wheat flour.

Claim 36

Dependent Claim 11 words

The method of claim 16 wherein the solution contains wheat flour.

Claim 37

Dependent Claim 20 words

The method of claim 16 further including the steps of adsorbing the desired fraction of PBAA on a flour carrier.

Claim 38

Dependent Claim 68 words

A method of fractionating modified alpha-amylase, wherein the modified alpha-amylase is prepared according to the method of claim 7, 10 or 12, comprising the steps of: adding the modified alpha-amylase to about 50% alcohol solution containing NaCl, increasing the alcohol to about 54.5% thereby precipitating an undesired fraction; separating the undesired fraction, and increasing the alcohol to about 61.5% thereby precipitating a desired fraction (54.5-61.5%) of modified alpha-amylase.

Claim 39

Dependent Claim 37 words

A dough comprising a combination of: flour; water; from 0.5-2.0% calcium stearate; and from 18-150 DU of purified alpha-amylase per about 800 grams of said flour, said purified alpha-amylase prepared according to claim 7, 10 or 12.

Claim 40

Dependent Claim 15 words

The dough of claim 32 wherein the purified alpha-amylase is present in from 45-150 DU.

Claim 41

Dependent Claim 15 words

The dough of claim 33 wherein the purified alpha-amylase is present in from 90-150 DU.

Claim 42

Dependent Claim 14 words

The dough of claim 34 wherein the purified alpha-amylase is present from 45-72 DU.

Claim 43

Dependent Claim 13 words

The dough of claim 32 wherein the calcium stearate concentration is from 0.5-1.0%.

Claim 44

Dependent Claim 13 words

The dough of claim 36 wherein the calcium stearate concentration is about 0.5%.

Claim 45

Dependent Claim 22 words

The dough of claim 32 wherein the calcium stearate concentration is about 1.0% and about 54 DU of purified enzyme is present.

Claim 46

Dependent Claim 22 words

The dough of claim 32 wherein the calcium stearate concentration is about 1.0% and from 90-100 DU or purified enzyme is present.

Claim 47

Dependent Claim 131 words

A dough comprising in combination: a predetermined amount of flour; chlorinated tap water; a predetermined amount of surfactant weighing not in excess of 2.0% of the weight of said predetermined amount of flour, wherein said surfactant is selected from the group consisting of sodium stearoyl-2-lactylate, calcium stearate, glycerol monostearate, calcium stearoyl-2-lactylate and succinylated monoglyceride; and bacterial alpha-amylase completely free of detectable protease activity introduced to said dough in a predetermined amount not in excess of 150 DU per 800 grams of said predetermined amount of flour on a carrier comprising a portion of said flour, wherein the amount of surfactant and alpha-amylase combined is sufficient to retard firming of the product made from said dough, and wherein said alpha-amylase is prepared according to the method of claim 7, 10 or 12.

Claim 48

Dependent Claim 115 words

A dough comprising in combination: a predetermined amount of flour; chlorinated tap water; a predetermined amount of surfactant weighing not in excess of 2.0% of the weight of said predetermined amount of flour, wherein said surfactant is selected from the group consisting of sodium stearoyl-2-lactylate, calcium stearate, glycerol monostearate, calcium stearoyl-2-lactylate and succinylated monoglyceride; and bacterial alpha-amylase completely free of detectable protease activity introduced to said dough in a predetemined amount not in excess of 150 DU per 800 grams of said predetermined amount of flour on a carrier comprising a portion of said flour wherein the amount of surfactant and alpha-amylase combined is sufficient to retard firming of the product made from said dough.

Claim 49

Dependent Claim 115 words

A dough comprising in combination: a predetermined amount of flour; chlorinated tap water; a predetermined amount of surfactant weighing not in excess of 2.0% of the weight of said predetermined amount of flour, wherein said surfactant is selected from the group consisting of sodium stearoyl-2-lactylate, calcium stearate, glycerol monostearate, calcium stearoyl-2-lactylate and succinylated monoglyceride; and bacterial alpha-amylase completely free of detectable protease activity introduced to said dough in a predetemined amount not in excess of 150 DU per 800 grams of said predetermined amount of flour on a carrier comprising a portion of said flour wherein the amount of surfactant and alpha-amylase combined is sufficient to retard firming of the product made from said dough.

Claim 50

Dependent Claim 24 words

The dough of claim 41 wherein said predetermined amount of alpha-amylase is about 54 DU per 800 grams of said predetermined amount of flour.

Claim 51

Dependent Claim 10 words

The dough of claim 41 wherein the alpha-amylase is fractionated.

Claim 52

Dependent Claim 13 words

The dough of claim 41 wherein the alpha-amylase is produced by Bacillus subtilis.

Claim 53

Dependent Claim 24 words

The dough of claim 41 wherein said predetermined amount of surfactant is present from 0.25-2.0% of the weight of said predetermined amount of flour.

Claim 54

Dependent Claim 25 words

The dough of claim 41 wherein said predetermined amount of alpha-amylase is present from 18-150 DU per 800 grams of said predetermined amount of flour.

Claim 55

Dependent Claim 12 words

The dough of claim 41 wherein said dough is a bread dough.

Claim 56

Dependent Claim 13 words

The dough of claim 41 wherein said dough is a yeast-raised donut dough.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1984-11-09	1985-05-11	1985-11-10	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1988-11-09	1989-05-11	1989-11-10	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1992-11-09	1993-05-11	1993-11-10	$7700.00	Window Closed / Lapsed

Modified enzyme system to inhibit bread firming method for preparing same and use of same in bread and other bakery products

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