A kind of soluble-insoluble ucst type pmaac carrier, its immobilized enzyme and application

A carrier and solubility technology, applied in the direction of immobilized enzymes, immobilized on/in organic carriers, glycosylases, etc., can solve the problems of difficult recovery and low efficiency of cellulose hydrolysis, and achieve improved stability and effective It is beneficial to full contact and solves the effect of LCST type low-efficiency hydrolysis problem

Active Publication Date: 2020-09-25
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the problem that the immobilized enzyme is difficult to recover, and the synthesized immobilized enzyme has low efficiency for cellulose hydrolysis, thereby synthesizing a soluble-insoluble high critical solution temperature (UCST) type carrier polymethacrylic acid sodium salt (PMAAc ) to immobilize cellulase, and co-immobilize cellulase and β-glucosidase

Method used

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  • A kind of soluble-insoluble ucst type pmaac carrier, its immobilized enzyme and application
  • A kind of soluble-insoluble ucst type pmaac carrier, its immobilized enzyme and application
  • A kind of soluble-insoluble ucst type pmaac carrier, its immobilized enzyme and application

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Effect test

Embodiment 1

[0050] Embodiment 1: the preparation of soluble-insoluble UCST type PMAAc carrier and the immobilization of enzyme

[0051] (1) Preparation of soluble-insoluble UCST PMAAc carrier:

[0052] At room temperature, dissolve 1 g of acrylic acid (AAc) and 0.022 g of ammonium persulfate (APS) in 10 mL of deionized water, then transfer them to a three-necked flask, and slowly add 0.32 g of methacrylamide dissolved in 10 mL of deionized water dropwise. (MAA), after shaking well, bubbling with nitrogen gas for about half an hour, then transfer to a single-necked flask, vacuumize under nitrogen protection, magnetically stir at 60 °C for 3.5 hours, take it out, cool and centrifuge, remove the supernatant, and use deionized water The temperature was raised to resuspend the material, and the product was obtained after repeated three times. After drying in a vacuum oven at 40° C. overnight, a transparent gel-like product was obtained. Next, lead an appropriate amount of PMAAc solution into ...

Embodiment 2

[0057] Embodiment 2: the preparation of soluble-insoluble UCST type PMAAc carrier and the immobilization of enzyme

[0058] (1) Preparation of soluble-insoluble UCST PMAAc carrier:

[0059] At room temperature, dissolve 1.88 g of acrylic acid (AAc) and 0.042 g of ammonium persulfate (APS) in 10 mL of deionized water, then transfer them to a three-necked flask, and slowly add 0.52 g of methacrylic acid dissolved in 10 mL of deionized water dropwise. Amide (MAA), after shaking well, bubbling with nitrogen gas for about half an hour, then transferred to a single-necked flask, vacuumed under nitrogen protection, magnetically stirred at 60 °C for 4.5 hours, then taken out, cooled, centrifuged, supernatant removed, and deionized The material was resuspended in water, and the product was obtained after repeated three times. After drying in a vacuum oven at 40° C. overnight, a transparent gel-like product was obtained. Next, lead an appropriate amount of PMAAc solution into a glass t...

Embodiment 3

[0064] Embodiment 3: the preparation of soluble-insoluble UCST type PMAAc carrier and the immobilization of enzyme

[0065] (1) Preparation of soluble-insoluble UCST PMAAc carrier:

[0066] At room temperature, 1.44 g of acrylic acid (AAc) and 0.032 g of ammonium persulfate (APS) were dissolved in 10 mL of deionized water, then transferred to a three-necked flask, and slowly added dropwise with methacrylamide dissolved in 10 mL of deionized water ( MAA, 0.42 g), shake well, then bubble with nitrogen for about half an hour, then transfer to a single-necked flask, vacuumize under nitrogen protection, stir magnetically at 60°C for 4 hours, take it out, cool and centrifuge, remove the supernatant, and use The material was resuspended in deionized water, and the product was obtained after repeated three times. After drying overnight in a vacuum oven at 40°C, a transparent gel-like product was obtained. Next, lead an appropriate amount of PMAAc solution into a glass test tube, hang...

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Abstract

The invention belongs to the technical field of immobilized enzymes, relates to a soluble-insoluble UCST type PMAAc vector as well as an immobilized enzyme and an application thereof, and in particular to a preparation method of the soluble-insoluble high critical solution temperature (UCST) type sodium polymethylacrylate (PMAAc) vector, an application of the immobilized enzyme and an applicationof an immobilized enzyme degraded cellulose material thereof. The preparation method comprises the following steps: dissolving acrylic acid and ammonium persulfate through de-ionized water; slowly dropping methacrylamide which is dissolved by de-ionized water; uniformly shaking an obtained solution, filling nitrogen until bubbling and transferring an obtained solution into a single-mouth flask; implementing vacuumizing in nitrogen protection; conducting a reaction under magnetic stirring; implementing centrifuging after conducting cooling; discarding supernatant liquid; and heating up a resuspended material via de-ionized water, so that the finished product (the PMAAc vector) is obtained. The vector provided by the invention is applicable to immobilized cellulase, and the vector is used for degrading cellulose materials; the immobilized enzyme prepared by the invention is excellent in storage stability and reuse effect; and efficiency of hydrolyzing and catalyzing the cellulose materials is also obviously improved.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, and relates to a UCST type PMAAc carrier, its immobilized enzyme and its application, in particular to a soluble-insoluble high critical solution temperature (UCST) type methacrylamide-acrylic acid copolymer (PMAAc ) carrier preparation method and application of immobilized cellulase and co-immobilized cellulase and β-glucosidase. Background technique [0002] Cellulose, one of the most abundant resources on earth, can be converted into functionally diverse glucose through chemical and biological means. At present, the fossil energy crisis is becoming more and more serious, and the green way of converting biomass glucose into fuel ethanol is becoming a major research hotspot in the energy industry. Because the free enzyme is difficult to recycle, and after a long time of enzymatic hydrolysis, the enzyme activity is easy to lose, these are the main obstacles inhibiting its industrial ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/18C12N11/087C12P19/14C12P19/02C08F220/06C08F220/56
CPCC08F220/06C12N9/2437C12N9/2445C12N11/08C12N11/18C12P19/02C12P19/14C12Y302/01021C08F220/56
Inventor 王赟韩娟王蕾李春梅倪良罗鹏万靖
Owner JIANGSU UNIV
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