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Method for Producing a Dairy Product

a technology for producing dairy products and products, applied in dairy products, milk preparations, buttermilk, etc., can solve the problems of low production of galactooligosaccharides and efficient hydrolysis, and achieve the effect of high efficiency, high amount of galactooligosaccharides, and high efficiency

Inactive Publication Date: 2010-11-11
NOVOZYMES AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009]The present inventors have surprisingly found that a C-terminally truncated fragment of the extracellular lactase from Bifidobacterium bifidum, which was originally isolated and patented for its ability to make high amounts of galactooligosaccharides from lactose, can be used very successfully for hydrolysis of lactose in milk. When tested in water+100 g / l lactose at 37° C., the enzyme makes galactooligosaccharides with high efficiency as described in the prior art. However, when tested in milk, the ratio of hydrolytic to transgalactosylating activity has changed markedly, resulting in efficient hydrolysis and very low production of galactooligosaccharides.
[0016]Further, the inventors have surprisingly found that very low levels of lactose can be achieved when using the lactase from Bifidobacterium bifidum as compared to other lactases typically used for treatment of milk. Another unexpected advantage of using the lactase from Bifidobacterium bifidum is that the enzyme is active at high temperatures, allowing for treatment of the milk at, e.g., 52° C., thus reducing the microbial count and thereby improving the quality of the milk.
[0026]Use of a lactase enzyme being active over a broad pH spectrum is especially useful for the production of fermented dairy products, where it allows for low enzyme dosage, since the enzyme is still active during and after fermentation. Also, very low levels of lactose in the fermented dairy product can be reached using such enzyme.

Problems solved by technology

However, when tested in milk, the ratio of hydrolytic to transgalactosylating activity has changed markedly, resulting in efficient hydrolysis and very low production of galactooligosaccharides.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0192]Lactase Activity-Assay in Eppendorf Tubes at 37° C., pH 6.5

[0193]Principle:

[0194]Lactase hydrolyses lactose into glucose and galactose. Glucose is measured after a modified version of the common glucose oxidase / peroxidase assay (Werner, W. et al. (1970) Z. analyt. Chem. 252: 224.).

[0195]LAU is defined as the amount of enzyme liberating 1 micromole of glucose per min at 37° C., pH 6.5 in M-buffer (M-buffer is defined in the description of the present patent application). Alternatively, the activity in LAU for a specific lactase may be determined by the method described here. The value obtained is compared to a standard curve run with a lactase of known activity, and the activity of the unknown sample calculated from this. The lactase of known activity may, e.g., be Lactozym obtained from Novozymes A / S, Denmark.

[0196]Solutions:

[0197]Assay buffer: 50 mM succinate, 50 mM HEPES, 50 mM CHES, 150 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 0.01% Triton X100, pH 6.5

[0198]GOD-Perid solution: 65 mM...

example 3

[0215]100 ml 15 or 30% (w / w) whey permeate containing primarily lactose and ions was made by mixing 15 or 30 g spray-dried whey permeate powder (Variolac, Arla) in 85 or 70 ml ionic water respectively. The solution was poured in a flask containing a magnetic stirring bar and placed in a water bath at 37° C. After 15 min, enzyme was added. Enzymes tested were Lactozym, a commercially available lactase from Novozymes A / S, Denmark, having an activity of 3060 LAU / g, and an experimental lactase from Bifidobacterium bifidum having the encoded sequence shown in SEQ ID NO: 2 and an activity of 295 LAU / g.

[0216]Dosages were 4225 LAU / l milk of Lactozym and 2025 LAU / l milk of the Bifidobacterium lactase. Milk samples were taken at regular intervals up till 5.5 hrs. and the enzyme inactivated by heating to 99° C. for 10 min in a thermomixer. Samples were diluted appropriately and filtered through a 0.20 um filter.

[0217]Lactose hydrolysis was measured using a Dionex BioLC equipped with a Dionex P...

example 4

[0220]pH Profile (at 37° C.) and Temperature Profile (at pH 6.5) of Experimental Lactase from Bifidobacterium bifidum using Lactose as Substrate.

[0221]Principle:

[0222]Lactase hydrolyses lactose and glucose+galactose is formed. Glucose is measured after a modified version of the common glucose oxidase / peroxidase assay (Werner, W. et al. (1970) Z. analyt. Chem. 252: 224.) pH profile

[0223]Substrate:

[0224]167 mM lactose, 50 mM succinate, 50 mM HEPES, 50 mM CHES, 150 mM KCl, 2 mM CaCl2, 1 mM MgCl2 and pH adjusted to pH 3, 4, 5, 6, 7, 8, 9 and 10 with NaOH.

[0225]Enzyme Sample:

[0226]Experimental lactase from Bifidobacterium bifidum having the encoded sequence shown in SEQ ID NO: 2 was diluted appropriately in 150 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 0.01% Triton X100.

[0227]Procedure:[0228]10 ul enzyme sample diluted in enzyme dilution buffer was added to PCR tubes at room temp.[0229]90 ul substrate was added at room temp. and quickly placed in a Peltier Thermal Cycler (PCT-200, MJ research) at ...

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PUM

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Abstract

The present invention relates to a method for producing a dairy product using an enzyme having lactase activity.

Description

SEQUENCE LISTING[0001]The present invention comprises a sequence listing.TECHNICAL FIELD[0002]The present invention relates to a method for producing a dairy product using an enzyme having lactase activity.BACKGROUND OF THE INVENTION[0003]Lactose intolerance is perhaps the best-known food sensitivity in the United States and other parts of the world. It is estimated that about 70% of the world's population has a genetically controlled limited ability to digest lactose. Therefore, to help dairy maldigesters keep dairy foods in their diet, there is a growing demand for dairy food products that contain no or only low levels of lactose.[0004]Lactase is used commercially to break down lactose in milk to produce dairy products which are suitable for people with lactose intolerance and / or have a sweeter taste. Because glucose and galactose are sweeter than lactose, lactase produces a more pleasant taste. Lactase is also used in the manufacture of ice cream. Lactose crystallises at the low ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A23C9/12A23C9/123
CPCA23C9/1206A23C9/123
Inventor HENDRIKSEN, HANNE VANGERNST, STEFFENWILTING, REINHARDTAMS, JEPPE WEGENERRUNGE, METTE OERHRSTROEMGULDAGER, HELLE SKOV
Owner NOVOZYMES AS
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