Preservation method of biological enzyme preparation

Abstract

The invention discloses a bio-enzyme preparation preserving method, which belongs to the technical field of biological engineering. The method preserves immobilized beta-galactosidase by washing and sterilizing immobilized beta-galactosidase and storing at lower temperature. For the immobilized beta-galactosidase which is used for decomposing lactose in milk, the method can enable the immobilizedenzyme to be repeatedly used for a long time, keep production continuity and lower production cost. Under a noncontinuous use condition, the adoption of the method can be effectively avoid microbial pollution and effectively retain the activity of the immobilized enzyme. When the method is used, the problems of low activity of the immobilized enzyme, influences on reaction speed and production efficiency and high production cost due to pollution of microbe to the immobilized beta-galactosidase in a use process and influences of proteins, fat and minerals are solved, and the reliable, effective and convenient immobilized enzyme washing, sterilizing and storing of lactose-decomposed milk and galactooligosaccharide-containing products can be realized.

Description

technical field

[0001] The invention relates to a method in the technical field of bioengineering, in particular to a method for preserving biological enzyme preparations. Background technique

[0002] β-galactosidase (β-galactosidase, EC3.2.1.23), scientific name β-galactoside galactohydrolase, is a white powder, tasteless and odorless, and becomes a light brown liquid after dissolution. β-galactosidase hydrolyzes galactosidic bonds under specific conditions, and can hydrolyze lactose into glucose and galactose that are easily absorbed by the small intestine. At the same time, β-galactosidase also has the function of transgalactoside, and transfers galactose to the galactose base of lactose to form a type of galactooligosaccharides containing 2 to 5 monosaccharide molecules. The molecular formula is (Gal) n-Glu, n=2-4. β-galactosidase hydrolyzes lactose and transglycosides as figure 1 shown.

[0003] Since β-galactosidase has the function of transgalactoside, it can gen...

Examples

Embodiment 1

[0040] Example 1: Application Results of Immobilized β-Galactosidase for Production of Lactose Decomposed Milk

[0041] Immobilization method of β-galactosidase: Weigh 2g chitosan beads → add 20ml 0.5% glutaraldehyde → cross-link for 40min (30°C, 80rpm) → wash to get activated chitosan beads → add 10ml of 1mg / ml enzyme solution, fix overnight → wash with water → store at 4°C.

[0042] Production of lactose-decomposed milk with immobilized β-galactosidase: use skim milk, after ultra-high temperature (140°C, 4s) sterilization treatment, determine the content of lactose, protein and fat, and cool it as the reaction substrate. Make the substrate milk pass through the reactor at a flow rate of 1L / h to react with the immobilized enzyme. After 4-5 cycles, the lactose is decomposed, the remaining lactose content is measured, and the lactose decomposition rate is calculated. When the decomposition of lactose reaches more than 80%, change to a new batch of substrate milk and operate a...

Embodiment 2

[0054] Example 2: Results of application of immobilized β-galactosidase to whey

[0055] The immobilization method of β-galactosidase is the same as that in Example 1.

[0056] Treat whey with immobilized β-galactosidase to produce galacto-oligosaccharides: Use desalted whey powder to prepare a whey solution with a lactose content of 10%, and after ultra-high temperature (140°C, 4s) sterilization, determine Lactose and protein content, cooled as reaction substrates. The substrate whey is passed through the reactor at a flow rate of 1 L / h to react with the immobilized enzyme, and after 4-5 cycles, lactose is decomposed to generate galacto-oligosaccharides. Using ion chromatography, using ion exchange and gradient elution methods, combined with pulsed amperometric detection system to detect lactose and galactooligosaccharides. When the decomposition of lactose reaches more than 80%, change to a new batch of substrate whey and then operate, continuous operation every day 12 hou...

Embodiment 3

[0059] Example 3: Application Results of Immobilized β-Galactosidase for Processing Lactose Solution

[0060] The immobilization method of β-galactosidase is the same as that in Example 1.

[0061] Process the lactose solution with immobilized β-galactosidase to produce galacto-oligosaccharides: use lactose to prepare a solution with a lactose content of 10%, after ultra-high temperature (140°C, 4s) sterilization, measure the lactose content, and cool it as Reaction substrate. The substrate whey is passed through the reactor at a flow rate of 1 L / h to react with the immobilized enzyme, and after 4-5 cycles, lactose is decomposed to generate galacto-oligosaccharides. Using ion chromatography, using ion exchange and gradient elution methods, combined with a pulsed amperometric detection system to detect lactose and galactooligosaccharides. When the decomposition of lactose reaches more than 80%, a new batch of substrate whey is used for operation, and the operation is continue...

technical field

[0001] The invention relates to a method in the technical field of bioengineering, in particular to a method for preserving biological enzyme preparations. Background technique

[0002] β-galactosidase (β-galactosidase, EC3.2.1.23), scientific name β-galactoside galactohydrolase, is a white powder, tasteless and odorless, and becomes a light brown liquid after dissolution. β-galactosidase hydrolyzes galactosidic bonds under specific conditions, and can hydrolyze lactose into glucose and galactose that are easily absorbed by the small intestine. At the same time, β-galactosidase also has the function of transgalactoside, and transfers galactose to the galactose base of lactose to form a type of galactooligosaccharides containing 2 to 5 monosaccharide molecules. The molecular formula is (Gal) n-Glu, n=2-4. β-galactosidase hydrolyzes lactose and transglycosides as figure 1 shown.

[0003] Since β-galactosidase has the function of transgalactoside, it can gen...

Examples

Embodiment 1

[0040] Example 1: Application Results of Immobilized β-Galactosidase for Production of Lactose Decomposed Milk

[0041] Immobilization method of β-galactosidase: Weigh 2g chitosan beads → add 20ml 0.5% glutaraldehyde → cross-link for 40min (30°C, 80rpm) → wash to get activated chitosan beads → add 10ml of 1mg / ml enzyme solution, fix overnight → wash with water → store at 4°C.

[0042] Production of lactose-decomposed milk with immobilized β-galactosidase: use skim milk, after ultra-high temperature (140°C, 4s) sterilization treatment, determine the content of lactose, protein and fat, and cool it as the reaction substrate. Make the substrate milk pass through the reactor at a flow rate of 1L / h to react with the immobilized enzyme. After 4-5 cycles, the lactose is decomposed, the remaining lactose content is measured, and the lactose decomposition rate is calculated. When the decomposition of lactose reaches more than 80%, change to a new batch of substrate milk and operate a...

Embodiment 2

[0054] Example 2: Results of application of immobilized β-galactosidase to whey

[0055] The immobilization method of β-galactosidase is the same as that in Example 1.

[0056] Treat whey with immobilized β-galactosidase to produce galacto-oligosaccharides: Use desalted whey powder to prepare a whey solution with a lactose content of 10%, and after ultra-high temperature (140°C, 4s) sterilization, determine Lactose and protein content, cooled as reaction substrates. The substrate whey is passed through the reactor at a flow rate of 1 L / h to react with the immobilized enzyme, and after 4-5 cycles, lactose is decomposed to generate galacto-oligosaccharides. Using ion chromatography, using ion exchange and gradient elution methods, combined with pulsed amperometric detection system to detect lactose and galactooligosaccharides. When the decomposition of lactose reaches more than 80%, change to a new batch of substrate whey and then operate, continuous operation every day 12 hou...

Embodiment 3

[0059] Example 3: Application Results of Immobilized β-Galactosidase for Processing Lactose Solution

[0060] The immobilization method of β-galactosidase is the same as that in Example 1.

[0061] Process the lactose solution with immobilized β-galactosidase to produce galacto-oligosaccharides: use lactose to prepare a solution with a lactose content of 10%, after ultra-high temperature (140°C, 4s) sterilization, measure the lactose content, and cool it as Reaction substrate. The substrate whey is passed through the reactor at a flow rate of 1 L / h to react with the immobilized enzyme, and after 4-5 cycles, lactose is decomposed to generate galacto-oligosaccharides. Using ion chromatography, using ion exchange and gradient elution methods, combined with a pulsed amperometric detection system to detect lactose and galactooligosaccharides. When the decomposition of lactose reaches more than 80%, a new batch of substrate whey is used for operation, and the operation is continue...

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