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Method for performing catalytic synthesis of starch phosphate monoester by using yeast display lipase

A technology for phosphate monoester and synthesizing starch, applied in the field of bioengineering, can solve the problems of complicated and time-consuming immobilization process, high production cost, limited commercial application, etc. Effect

Inactive Publication Date: 2014-07-16
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Lipase is currently the most widely used enzyme in esterification synthesis, but its commercial application is greatly limited by the high production cost and complicated and time-consuming immobilization process.

Method used

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  • Method for performing catalytic synthesis of starch phosphate monoester by using yeast display lipase
  • Method for performing catalytic synthesis of starch phosphate monoester by using yeast display lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Preparation of yeast display cell lipase

[0018] Synthesize the lipase gene (Genbank number: AF229435) of Rhizopus oryzae and the cell wall α-lectin gene of Pichia pastoris GS115 (Genbank number: M28164) by artificial synthesis, and add Connect the peptide sequence GSSGGSGGSGGSGGSGS(linker), and get the nucleotide sequence pro-ROL-linker-α-agglutinin after connection, and add EcoR I and Not I restriction sites at both ends of the sequence, where pro-ROL is the lipase gene , α-agglutinin is the cell wall α lectin gene.

[0019] Using the above artificially synthesized sequence as a template, PCR amplification was performed using the following primer pair,

[0020] Upstream primer: 5'-AAGGAAAAAAAGAATTCGTTCCAGTTTCTGG-3';

[0021] Downstream primer: 5'-TTTTCCTTTTGCGGCCGCTAATGAAACG-3'

[0022] The PCR reaction system is: 1 μl of template DNA, 0.5 μl of high-fidelity DNA polymerase, 0.4 μl of dNTP (50 mM), 0.5 μl of upstream and downstream primers, 5 μl of 10×PC...

Embodiment 2

[0026] Example 2 yeast display lipase catalyzed synthesis of starch phosphate monoester

example 1

[0027] Example 1 Take 350g corn starch and prepare 35% starch milk (starch dry base) with water, absorb water at 65°C for 15min to make the starch milk absorb water and swell, add 1g of the above-mentioned prepared yeast to display lipase after cooling, and then add 10g NaH in batches 2 PO 4 2H 2 O and Na 2 HPO 4 12H 2 O (weight ratio 3: 1) mixture, placed in a 85-1 type magnetic stirrer to stir and start the reaction, the rotating speed is 200 rpm, the reaction temperature is maintained at 60 ° C, the stirring is stopped after 1 hour of reaction, and the pH value is adjusted. 7.0, centrifuge to take the precipitate, wash the precipitate with water several times, and spray dry.

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Abstract

The invention discloses a method for performing catalytic synthesis of starch phosphate monoester by using yeast display lipase. The method comprises the following steps: dissolving cereal starch in water, adding NaH2PO4.H2O, Na2HPO4.12H2O and yeast display lipase after imbibition, reacting at 55-65 DEG C for 1-1.5 hours under stirring, separating, and purifying to prepare starch phosphate monoester. In the method, lipase is displayed outside cells and the biocatalyst is utilized to perform the catalytic synthesis of starch acetate, thus the conversion efficiency can be increased, the reaction time can be shortened and the production cost can be reduced.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for synthesizing starch phosphate monoester catalyzed by yeast surface lipase. Background technique [0002] Starch phosphate monoester is a kind of starch derivative produced by the esterification of native starch and phosphate. It is an anionic modified starch. Compared with native starch, starch phosphate monoester has transparent dispersion, high viscosity, anti-aging and stable Good properties and good water retention performance, when stored at low temperature for a long time or repeated freezing and thawing, there is no water analysis. Starch phosphate monoesters with different degrees of substitution have been widely used in food, medicine, chemical and other industries. Especially in food technology, as a food additive, starch phosphate monoester with low and medium substitution degree is widely used in the processing of frozen food and other aspects. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/04C12N15/63C12N1/19C12N9/20C12R1/84
Inventor 阮晖王睿之古再丽努尔徐娟周陈伟林吉恒何国庆
Owner ZHEJIANG UNIV
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