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Dry powder type immobilized lipase and production process thereof

A lipase and immobilized carrier technology, applied in the field of biomedicine, can solve the problem of lack of immobilized pig liver esterase

Active Publication Date: 2018-03-20
ABIOCHEM BIOTECHNOLOGY (CHONGQING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is still a lack of immobilized pig liver esterase that can be used in the organic phase in the art

Method used

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  • Dry powder type immobilized lipase and production process thereof
  • Dry powder type immobilized lipase and production process thereof
  • Dry powder type immobilized lipase and production process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Dissolve porcine liver lipase (porcine liver esterase, purchased from Sigma, 15kU / g) in 50mM phosphate buffer (pH 7.5, specific enzyme activity of enzyme solution is 100u / mL), according to the ratio of 20g resin / 100mL enzyme solution Join Lifetech TM ECR1030M resin (also known as ECR1030M resin, purchased from Purolite, with a water content of about 70%, the same below), was subjected to immobilization reaction. The immobilization conditions were 16°C, 120 rpm, stirring for 16 hours, and standing for 4 hours. The immobilized enzymes were collected and washed with tap water and 50 mmol / L, pH 7.5 phosphate buffer solution to remove impurities on the resin surface and weakly adsorbed enzymes. After vacuum filtration, samples were taken to measure moisture and enzyme activity respectively, the moisture content was 69%, the enzyme activity was 500u / g, and the enzyme activity yield was 100%. The water-containing immobilized enzyme obtained is further dried with an ebullatin...

Embodiment 2

[0069] Dissolve porcine liver lipase (purchased from Sigma, 15kU / g) in 50mM phosphate buffer (pH 7, specific enzyme activity of the enzyme solution is 100u / mL), add Lifetech at a ratio of 15g resin / 100mL TM ECR1030M resin, for immobilization reaction. The immobilization conditions were 10°C, 150 rpm, stirring for 20 hours, and standing for 4 hours. The immobilized enzymes were collected and washed with tap water and 50mmol / L, pH7.5 phosphate buffer solution to remove impurities on the resin surface and weakly adsorbed enzymes. After vacuum filtration, samples were taken to measure water content and enzyme activity, the water content was 69%, the enzyme activity was 550u / g, and the enzyme activity yield was 82.5%. The obtained water-type immobilized enzyme was dried with a fluidized bed, the temperature of the air inlet of the fluidized bed was 35°C, the temperature of the air outlet was 25°C, and the drying time was 20min to obtain 8.5g of immobilized enzyme with a moisture c...

Embodiment 3

[0072] Dissolve porcine liver lipase (purchased from Sigma, 15kU / g) in 50mM phosphate buffer (pH 7.5, the specific enzyme activity of the enzyme solution is 100u / mL), add Lifetech at a ratio of 18g resin / 100mL TM ECR1030M resin, for immobilization reaction. The immobilization conditions were 15°C, 100 rpm, stirring for 20 hours, and standing for 5 hours. The immobilized enzymes were collected and washed with tap water and 50mmol / L, pH7.5 phosphate buffer solution to remove impurities on the resin surface and weakly adsorbed enzymes. After vacuum filtration, 18.5 g of wet immobilized enzyme was obtained, the water content was 69%, the enzyme activity was 530u / g, and the enzyme activity yield was 95.4%. The obtained water-containing immobilized enzyme is dried with a fluidized bed, the temperature of the air inlet of the fluidized bed is 40°C, the temperature of the air outlet is 25°C, and the drying time is 15min. The measured moisture content is 45%, and the enzyme activity i...

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Abstract

The invention provides dry powder type immobilized lipase and a production process thereof and particularly provides dry powder type immobilized pig live lipase. The dry powder type immobilized pig live lipase contains pig liver lipase and immobilized carrier resin, wherein the pig liver lipase is adhered to the immobilized carrier resin; and the water content (weight) of the dry powder type immobilized pig live lipase is 40%-50%. The invention further provides a preparation method of the dry powder type immobilized pig live lipase. The immobilized pig live lipase can present extremely high catalytic activity in an organic phase, is good in stability and can be repeatedly used.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular, the invention relates to a dry powder immobilized lipase and a production process thereof. Background technique [0002] Lipase is an enzyme widely used in the field of medicine and chemical industry, and can be used for the resolution of chiral acid, chiral amine and chiral alcohol pharmaceutical intermediates. Pig liver esterase is a lipase found in pig liver. The production of the antihypertensive drug Aliskerin is to use pig liver lipase to split chiral acid; it is also used in the split of chiral diacids, diols, and diamines. Excellent performance. [0003] Immobilized enzymes refer to enzymes that are locked in a certain space and can react continuously, and the reacted enzymes can be recycled and reused. For example, the immobilized Candida antarctica lipase CalB represented by Novo 435 can be used in the organic phase. Compared with the free enzyme, it has the following advanta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/20C12N11/08C12N9/98
CPCC12N9/20C12N9/98C12N11/08
Inventor 田振华罗煜丁时诚瞿旭东
Owner ABIOCHEM BIOTECHNOLOGY (CHONGQING) CO LTD
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