Co-crosslinking immobilization method of porcine pancreatic lipase

A porcine pancreatic lipase, co-crosslinking technology, applied in immobilized enzymes, biochemical equipment and methods, immobilized on/in organic carriers, etc., can solve the problem of high reactivity, excessive crosslinking of enzymes, loss of enzyme activity and other problems, to achieve the effect of improving the catalytic activity and improving the microenvironment

Active Publication Date: 2020-02-07
NINGBO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The most commonly used cross-linking agent is water-soluble glutaraldehyde, which has high reactivity and is difficult to control the dosage. It is easy to cause excessive cross-linking of the enzyme and cause a great loss of enzyme activity. In addition, the tr

Method used

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  • Co-crosslinking immobilization method of porcine pancreatic lipase
  • Co-crosslinking immobilization method of porcine pancreatic lipase
  • Co-crosslinking immobilization method of porcine pancreatic lipase

Examples

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Embodiment Construction

[0015] Enzyme immobilization

[0016] 1) Bisphenol A epoxy resin (brand name is E-44, epoxy value is 0.44, number average molecular weight is 454), three kinds of components of methanol and diethylenetriamine are mixed according to the mass ratio of 2: 2: 1, Stir and react in the range of 25-35°C for 4-5 hours, pour the mixture into water, wash the precipitate repeatedly with water to remove methanol and a small amount of amine, then put it in a vacuum oven and dry it at room temperature to obtain an aminated epoxy resin;

[0017] 2) Add epoxy resin amides and β-cyclodextrin into water at a molar ratio of 1:2.1 to 1:2.3, heat and stir until all epoxy resin amides are converted into molecular complexes and dissolved in water, and keep the The total mass concentration of the aqueous solution is in the range of 5 to 6wt.%.

[0018] 3) Dissolving porcine pancreatic lipase in a phosphate buffer solution with pH=7.0, keeping the enzyme concentration in the range of 1.0-7.0 mg / mL; ...

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Abstract

The invention relates to a co-crosslinking immobilization method of porcine pancreatic lipase. Oil-soluble butanediol diacrylate is used as a cross-linking agent, reactants in an aqueous phase are amino-containing porcine pancreatic lipase and a supermolecular complex formed by aminated epoxy resin and beta-cyclodextrin, through a Michael addition reaction of dual bonds and amino, a co-crosslinking polymerization reaction is generated at a low temperature, and immobilized porcine pancreatic lipase of different loading capacities can be prepared. The cross-linking degree is controlled, so thatthe dispersibility is improved, and internal mass transfer microenvironment can be improved. The immobilized enzyme has high catalytic activity, when the loading capacity is 42mg of enzymes for 1g ofcarriers, the activity achieves a highest value.

Description

technical field [0001] The invention relates to the technical field of immobilized enzyme biocatalysis, in particular to a method for co-crosslinking and immobilizing porcine pancreatic lipase. The novel immobilized enzyme can be used for hydrolysis of fats and also for dynamic resolution of chiral compounds. Background technique [0002] Porcine pancreatic lipase (EC 3.1.1.3), also known as triacylglycerol hydrolase, is an important acyl hydrolase that can catalyze the hydrolysis of triglycerides to produce diglycerides, monoglycerides, glycerol and fatty acids. Its molecular weight is between 45,000 and 50,000 Da, and it is an acidic protein (the isoelectric point is 5.0). Structurally, porcine pancreatic lipase is ellipsoidal, with a three-dimensional size of 4.6nm×2.6nm×1.1nm. It contains a large number of hydrophobic amino acids, which can form hydrophobic regions, and its polarity index is 47. In terms of composition, except for isoleucine which is different from othe...

Claims

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Application Information

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IPC IPC(8): C12N11/096C12N11/10
CPCC12N11/08C12N11/10C12N9/20C12Y301/01003Y02P20/50
Inventor 吴嘉沁张瑞丰李艳肖通虎龙能兵
Owner NINGBO UNIV
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