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Detection method of transesterification activity of non-aqueous phase lipase

A detection method, lipase technology, which is applied in the measurement of color/spectral characteristics and analysis by making materials undergo chemical reactions, can solve the problems of easy volatilization of acetaldehyde, interference of detection results, difficulty in calculating residual content of reaction solution, etc., to achieve Low raw material cost, less time-consuming detection, simple and feasible detection method

Inactive Publication Date: 2013-09-18
ZHEJIANG UNIV OF TECH
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Problems solved by technology

The reaction solvent, the acetaldehyde obtained by the isomerization of the substrate alkyl alcohol and the product vinyl alcohol all absorb at 200nm, so they will all interfere with the detection results. Among them, acetaldehyde is easy to volatilize, and the residual content in the reaction solution is difficult to calculate

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  • Detection method of transesterification activity of non-aqueous phase lipase
  • Detection method of transesterification activity of non-aqueous phase lipase
  • Detection method of transesterification activity of non-aqueous phase lipase

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Embodiment 1

[0028] Example 1: Determination of transesterification activity of 7 commercial lipases and gas chromatography detection and verification of full-wavelength scanning diagrams and wavelengths of nitrophenol palmitate (pNPP) and p-nitrophenol (pNP) using ethanol as a solvent 310nm standard curve see Figure 1~4 .

[0029] 2ml n-hexane reaction system, pNPP concentration 10mM, isopropanol concentration 1M, 0.01g lipase, 30°C, 200r / min shaker speed for 10min, sample 30μl reaction supernatant diluted 100 times with ethanol and measure OD value at 310nm. The conversion rate of the reaction was calculated according to the standard curve of p-nitrophenol, and the enzyme activity data of lipase was obtained.

[0030] Conversion rate calculation formula: where A 0 、A 1 is the OD of blank control and sample detection 310nm , ε 1 , ε 2 are the molar extinction coefficients of pNPP and pNP3 at 10 nm.

[0031] At the same time, samples were diluted with ethyl acetate and analyzed b...

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Abstract

The invention provides a detection method of transesterification activity of non-aqueous phase lipase. The method comprises the following steps of: taking p-nitrophenol ester as shown in formula (I) and alkyl alcohol as shown in formula (II) as substrates, taking the lipase as a catalyst for performing transesterification in an organic solvent to generate p-nitrophenol and alkyl ester as shown in formula (III), performing organic dissolution and dilution, and then detecting the variation of an ultraviolet absorption value of a reaction solution at 310nm to determine the transesterification activity of the lipase. The beneficial effects of the detection method of the transesterification activity of the non-aqueous phase lipase, disclosed by the invention, are mainly represented as follows: the detection method is simple and feasible, the cost of raw materials is low, the time consumption in detection is low, the requirements for instruments are not high, and the detection method can be used for fast determination of esterification activity of the lipase, and is suitable for being applied to conventional biochemical laboratories.

Description

(1) Technical field [0001] The invention relates to a method for detecting transesterification activity of non-aqueous lipase. (2) Background technology [0002] Lipase (Lipase, EC3.1.1.3) is triacylglycerol acyl hydrolase, which catalyzes the hydrolysis of natural substrate oil to generate fatty acid, glycerol and monoglyceride or diglyceride, and can also catalyze esterification, transesterification, esterification Exchange and aminolysis reactions. The active center of most lipases is a triplet composed of serine, aspartic acid and histidine. The lipase molecule consists of a hydrophilic part and a hydrophobic part, and the active center is close to the hydrophobic end of the molecule. Lipase has a special cap structure and the property of interfacial activation. With the rapid development of non-aqueous biocatalysis in the past 20 years, there are more and more basic theoretical and applied researches on lipase. Lipase has been widely used in the fields of food, medi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/75G01N21/33
Inventor 郑建永汪钊付显锋
Owner ZHEJIANG UNIV OF TECH
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