A kind of cellulase suitable for preparing cellulose nanocrystal and its application

A technology of cellulase and nanocrystals, which is applied in the biological field to achieve the effects of small particle size, high hydrolysis efficiency, and accelerated fracture

Active Publication Date: 2022-05-03
SHANDONG UNIV
View PDF9 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is no report about the cellulase suitable for preparing cellulose nanocrystals and the method of obtaining the cellulase by fermentation and rapidly preparing cellulose nanocrystals by constructing an engineering strain with a reasonable enzyme system

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
  • A kind of cellulase suitable for preparing cellulose nanocrystal and its application
  • A kind of cellulase suitable for preparing cellulose nanocrystal and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] The construction of embodiment 1 engineering strain cEES

[0064] (1) Gene acquisition:

[0065]The activation of the cellulase gene (cbh1) (GenBank: GQ844299.1) was amplified with primers cbh1-U-F and cbh1-U-R using Penicillium oxalicum CGMCC No.5302 genome (GenBank: AGIH00000000.1) as a template Subsequence cbh1 (U), amplified with primers cbh1-D-hphR-F and cbh1-D-R to obtain the downstream sequence cbh1 (D) of the cellulase gene (cbh1) coding region, with primers Cel7B-cbh1 (U) R-F and Cel7B-hphF-R amplifies the gene fragment cel7B (GenBank: EU339127.1), and uses primers cel5B-F and cel5B-R to amplify the gene fragment cel5B (GenBank: KJ 955479); with Trichoderma reesei The cDNA of QM6a ATCC13631 (GenBank: AAIL00000000.2) was used as a template to amplify the gene swo1 (GenBank: AJ245918.1) with primers swo1-AgpdA(p)R-F and swo1-R; the plasmid pAN7-1 (GenBank: Z32698.1 ) was used as a template to amplify the promoter AgpdA( p)(GenBank:Z32524.1); the glyceraldehyde...

Embodiment 2

[0102] After the engineering strain cEES is activated, the seed liquid is obtained. Under sterile conditions, the hyphae are added to the fermentation medium in a proportion of 0.3% by weight, and cultured at 28°C and 250rpm for 7 days or in a fermenter for 7 days; Obtain the supernatant by centrifugation (10000rpm, 10min), which is the cellulase-containing fermentation broth. In the cellulase liquid, the ratio of endo-cellulase activity to exo-enzyme activity is 40, and the protein ratio of endo-cellulase Cel7B to Cel5B is 1.

[0103] Among them: the seed medium formula is: 20g / L glucose, Vogel's salt; the fermentation medium formula is: 10g / L bran, 10g / L microcrystalline cellulose, 3g / L KH 2 PO 4, 2.6g / L NaNO 3 , 0.5g / LMgSO 4 ·7H 2 O, 0.5g / L CaCl 2 , 1g / Lpeptone, 7.5mg / L FeSO 4 ·7H 2 O, 2.5mg / L MnSO 4 ·H 2 O, 3.6mg / L ZnSO 4 ·7H 2 O, 3.7mg / LCoCl 2 ·6H 2 O.

[0104] The formula of the above-mentioned Vogel's salt has been recorded and publicized in Chinese patent...

Embodiment 3

[0106] After the engineering strain cEES is activated, the seed liquid is obtained. Under sterile conditions, the hyphae are added to the fermentation medium in a proportion of 2% by weight, and cultivated at 35°C and 150rpm for 3 days or in a fermenter for 3 days; Obtain the supernatant by centrifugation (10000rpm, 10min), which is the cellulase-containing fermentation broth. In the cellulase liquid, the ratio of endo-cellulase activity to exo-enzyme activity was 44, and the protein ratio of endo-cellulase Cel7B to Cel5B was 1.1.

[0107] Among them: the seed medium formula is: 20g / L glucose, Vogel's salt; the fermentation medium formula is: 10g / L bran, 10g / L microcrystalline cellulose, 3g / L KH 2 PO 4 , 2.6g / L NaNO 3 , 0.5g / LMgSO 4 ·7H 2 O, 0.5g / L CaCl 2 , 1g / Lpeptone, 7.5mg / L FeSO 4 ·7H 2 O, 2.5mg / L MnSO 4 ·H 2 O, 3.6mg / L ZnSO 4 ·7H 2 O, 3.7mg / LCoCl 2 ·6H 2 O.

[0108] The formula of the above-mentioned Vogel's salt has been recorded and publicized in Chinese p...

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

PropertyMeasurementUnit
sizeaaaaaaaaaa
sizeaaaaaaaaaa
sizeaaaaaaaaaa
Login to view more

Abstract

The invention discloses a cellulase suitable for preparing cellulose nanocrystals and an application thereof, wherein the cellulase is prepared by the following steps: (1) construction of an engineering strain; (2) production of cellulase by liquid fermentation . In the process of hydrolyzing cellulose, the cellulase of the present invention strengthens the enzyme component that degrades the amorphous region of cellulose and the protein component that depolymerizes the hydrogen bonds in the large-sized cellulose crystal region, thereby promoting the formation of cellulose macromolecular chains. Fragmentation and "peeling" reaction of cellulose crystallization area, so it is easier to form nanoscale cellulose crystals by using the cellulase of the present invention. Compared with the existing commercial cellulase, the enzyme system composition of the cellulase of the present invention is more suitable for the preparation of cellulose nanocrystals, and has the advantages of controllable enzyme system, high hydrolysis efficiency, low enzyme cost, simple preparation process, It has the advantages of not needing to carry out mechanical treatment on the raw material, and the yield of nanocellulose is high.

Description

technical field [0001] The invention relates to a cellulase and an application thereof, in particular to a cellulase suitable for preparing cellulose nanocrystals and an application thereof, and belongs to the field of biotechnology. Background technique [0002] Cellulose nanocrystals have attracted extensive attention in recent years due to their renewable, nontoxic, high specific surface area, excellent mechanical properties, and easy functionalization of hydroxyl groups. Cellulose nanocrystals have the potential to develop a variety of products that are expected to replace traditional petrochemical and non-biodegradable polymer materials, including food packaging, films, and nanocellulose silver-loaded agents with antibacterial properties. [0003] The preparation methods of cellulose nanocrystals include concentrated acid hydrolysis and biological methods. Among them, the hydrolysis of concentrated sulfuric acid is the most commonly used preparation method at present, ...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12N15/56C12N15/81C12P19/14C12P19/04C12R1/80
CPCC12N9/2437C12N15/81C12P19/14C12P19/04
Inventor 赵建杨甜甜李雪芝
Owner SHANDONG 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