Method for catalyzing and synthesizing starch acetate through yeast show lipase

A technology of starch acetate and lipase, which is applied in the field of bioengineering, can solve the problems of complicated and time-consuming immobilization process, high production cost, and limited commercial application, and achieve the advantages of improved operation stability, low production cost, and shortened reaction time Effect

Inactive Publication Date: 2011-10-12
ZHEJIANG UNIV
View PDF0 Cites 4 Cited by
  • 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 applicatio

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for catalyzing and synthesizing starch acetate through yeast show lipase
  • Method for catalyzing and synthesizing starch acetate through yeast show lipase

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0017] Example 1 Preparation of yeast display lipase

[0018] Synthesize the lipase gene of Rhizopus oryzae (Genbank No.: AF229435) and the cell wall alpha lectin gene of Pichia pastoris GS115 (Genbank No. M28164) by artificial synthesis, and add the C-terminal of the lipase gene The peptide sequence GSSGGSGGSGGSGGSGS (linker) is connected to the upper part, and the nucleotide sequence pro-ROL-linker-α-agglutinin is obtained after the connection. At the same time, EcoR I and Not I restriction sites are added 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 pairs,

[0020] Upstream primer: 5’-AAGGAAAAAAGAATTCGTTCCAGTTTCTGG-3’;

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

[0022] The PCR reaction system is: 1μl template DNA, 0.5μl high-fidelity DNA polymerase, 0.4μl dNTP (50mM)...

Example Embodiment

[0026] Example 2 Yeast display lipase to catalyze the synthesis of starch acetate

Example Embodiment

[0027] Example 1 Prepare 35% starch milk (starch dry basis) from 350g of cereal starch with water, and treat it at 65°C for 15 minutes to absorb water and expand the starch milk. After cooling, add 1g of the yeast display lipase prepared above, and then add 10g of acetic acid in batches. The anhydride is placed in a 85-1 magnetic stirrer and stirred to start the reaction. The speed is 100 revolutions per minute. The reaction temperature is kept at 66°C. After 2 hours of reaction, the stirring is stopped, the pH is adjusted to 6.0, the precipitate is centrifuged to take the precipitate, and washed with water several times Precipitate, wash away unreacted acetic anhydride, then dry and pulverize the precipitate to obtain the finished product.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for catalyzing and synthesizing starch acetate through yeast show lipase, comprising the following steps of: dissolving grain starch in water; after imbibition, adding acetic anhydride and the yeast show lipase; stirring and reacting at 65-75 degrees centigrade for 2-2.5 h; and separating and purifying to obtain starch acetate. A method for preparing the yeast show lipase comprises the following steps of: transforming linearly treated recombinant plasmids into pichia pastoris GS115; inoculating the obtained transformant into a BMMY (buffered methanol-complex medium) culture medium; after inducing and culturing for 72-144 h, centrifugally collecting thallus; and washing, biologically imprinting, freezing and drying the thallus so as to obtain the yeast show lipase. By showing the lipase outside cells, the starch acetate is catalyzed and synthesized through the lipase preparation. By means of the method disclosed by the invention, the transformation efficiency is improved; furthermore, the reaction time is shortened; and the production cost is reduced.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for synthesizing starch acetate catalyzed by yeast display lipase. Background technique [0002] Starch acetate is an important type of modified starch, which can produce products with better coagulation resistance, transparency and other performance than native starch. Compared with native starch, starch acetate after acetylation shows the following properties Features: easy gelatinization, low gelatinization temperature, stable viscosity, neutral solution, no gel formation even after cooling, good film-forming property, greatly improved thermal stability, and greatly improved comprehensive performance , Has been used in food, paper, textile, adhesives and other industrial sectors. [0003] Compared with the chemical synthesis of starch acetate, the biological enzymatic method can obtain a reaction product with a high degree of substitution in a short period of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap