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A method of improving catalyzed resolution properties of free lipase in an organic phase

A technology of free lipase and organic phase, applied in the field of non-aqueous phase lipase catalytic chiral separation, to achieve low cost, low catalytic performance and low catalytic efficiency

Inactive Publication Date: 2016-04-20
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the effect of biosurfactant bile salts on improving the efficiency of lipase-catalyzed resolution in non-aqueous media.

Method used

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  • A method of improving catalyzed resolution properties of free lipase in an organic phase
  • A method of improving catalyzed resolution properties of free lipase in an organic phase
  • A method of improving catalyzed resolution properties of free lipase in an organic phase

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Effect test

Embodiment 1

[0029] Embodiment 1: a kind of method that improves free lipase catalytic resolution performance in organic phase, comprises the following steps:

[0030] S1, take a certain mass of sodium cholate and dissolve it in a phosphate buffer solution with pH=7.5, prepare a sodium cholate concentration of 16mM, and dilute the sodium cholate buffer solution with a concentration of 16mM to a solubility of 2mM, 3mM, 4mM, 6mM, 8mM sodium cholate phosphate buffer solution;

[0031] Weigh 5 parts of 0.01g porcine pancreatic lipase (PPL) or Candida columnar lipase (CRL) coarse powder and place them in 5 sample bottles in turn, add 5 different concentrations of sodium cholate to the 5 samples respectively Phosphate buffer solution 2ml;

[0032] Weigh 0.01g of porcine pancreatic lipase (PPL) or Candida columnar lipase (CRL) coarse powder and place it in a sample bottle and add 2ml of phosphate buffered saline solution as blank control group 1;

[0033] Pipette 2ml of phosphate buffer solutio...

Embodiment 2

[0042] Embodiment 2: a method for improving the catalytic resolution performance of free lipase in the organic phase, comprising the following steps:

[0043] S1, weigh a certain amount of sodium deoxycholate and dissolve it in a phosphate buffer solution with a pH of 7.5, prepare a concentration of sodium deoxycholate of 16 mM, and dilute the sodium deoxycholate buffer solution with a concentration of 16 mM in sequence 2mM, 3mM, 4mM, 6mM, 8mM sodium deoxycholate phosphate buffer solution;

[0044] Weigh 5 parts of 0.01g porcine pancreatic lipase (PPL) or Candida columnar lipase (CRL) coarse powder and place them in 5 sample bottles in turn, add 5 different concentrations of deoxycholic acid to the 5 samples respectively Sodium phosphate buffer solution 2ml;

[0045] Weigh 0.01g of porcine pancreatic lipase (PPL) or Candida columnar lipase (CRL) coarse powder and place it in a sample bottle and add 2ml of phosphate buffered saline solution as blank control group 1;

[0046] ...

Embodiment 3

[0055] Embodiment 3: a method for improving the catalytic resolution performance of free lipase in the organic phase, comprising the following steps:

[0056] S1, weigh a certain amount of sodium chenodeoxycholate and dissolve it in a phosphate buffer solution with pH = 7.5, prepare a concentration of sodium chenodeoxycholate of 16 mM, and buffer The solution was successively diluted into sodium chenodeoxycholate phosphate buffer solution with a solubility of 2mM, 4mM, 6mM, 8mM and 10mM;

[0057] Weigh 5 parts of 0.01g porcine pancreatic lipase (PPL) or Candida columnar lipase (CRL) coarse powder and place them in 5 sample bottles in turn, add 5 different concentrations of goose deoxygenate to the 5 samples respectively Sodium cholate phosphate buffer solution 2ml;

[0058] Weigh 0.01g of porcine pancreatic lipase (PPL) or Candida columnar lipase (CRL) coarse powder and place it in a sample bottle and add 2ml of phosphate buffered saline solution as blank control group 1;

...

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Abstract

The invention relates to a method of improving catalytic chiral alcohol resolution properties of free lipase in an organic phase by inducing lipase conformational changes by utilizing a bile salt. The method includes S1) weighing a catalytic amount of the lipase, and adding into a buffer solution containing the bile salt, S2) fully mixing the mixture in the step S1), performing vacuum freeze drying and grinding into powder, S3) mixing the powder prepared in the step S2) with an organic solvent, a chiral alcohol and vinyl acetate or isopropenyl acetate and reacting, and S4) after the reaction is finished, diluting the reaction solution, and detecting generation amounts of R-chiral alcohol and S-chiral alcohol through a high performance liquid chromatograph to determine chiral resolution activity of the lipase treated with the bile salt. The method is convenient in material preparation, simple in operation, good in repeatability and obvious in effects. Compared with lipase not subjected to treatment with the bile salt, the catalytic resolution properties of the lipase treated with the bile salt are greatly improved.

Description

technical field [0001] The invention belongs to the technical field of non-aqueous phase lipase catalyzed chiral separation, and in particular relates to a method for inducing and activating lipase by using bile salts to improve the catalytic resolution performance of free lipase in an organic phase. Background technique [0002] Lipase is a biological catalyst that exhibits excellent stereospecificity in the catalytic resolution process. However, the active region of lipase contains a "lid" structure, so as the surrounding environment of lipase changes, its "lid" will change to varying degrees. This will change the catalytic performance of lipase to a certain extent. [0003] Previously, some researchers used various surfactants to induce the activation of the active center of lipase by using the "lid" feature of lipase to enhance the catalytic activity of lipase. For example, Fernando et al. used the surfactant Tritonx-100 to stimulate the active site of BTL2 lipase, caus...

Claims

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Application Information

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IPC IPC(8): C12P41/00C12P7/22C12P7/62
Inventor 叶鹏胡鹏程韩祝平
Owner ZHEJIANG SCI-TECH UNIV
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