Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mutant of cellulosic exoglucanase cbh I and viscosity reducer of syrup in corn fuel ethanol fermentation production

A technology of exo-glucanase and cellulose, applied in the directions of fermentation, biofuels, enzymes, etc., can solve the problems of increasing energy consumption of feed drying steam, dark color of DDGS, destruction of nutrients, etc., and achieve excellent enzyme stability and catalytic activity, reducing steam consumption, and improving the effect of primary product yield

Inactive Publication Date: 2017-11-14
NANJING TIANKAI BIOTECH
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the concentration of syrup is not high, a large amount of water will be brought in, which will greatly increase the energy consumption of feed drying steam, increase the drying temperature, prolong the high temperature residence time, destroy a large amount of nutrients, produce a large amount of Maillard reaction, and make DDGS color It is darker and easily has a burnt smell, which affects the palatability of raised animals, affects the grade and price of DDGS feed, and thus affects the profits of corn fuel ethanol enterprises
[0005] In the prior art, there is no method for low energy consumption, rapid and efficient viscosity reduction of syrup in corn fuel ethanol fermentation production

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
  • Mutant of cellulosic exoglucanase cbh I and viscosity reducer of syrup in corn fuel ethanol fermentation production
  • Mutant of cellulosic exoglucanase cbh I and viscosity reducer of syrup in corn fuel ethanol fermentation production
  • Mutant of cellulosic exoglucanase cbh I and viscosity reducer of syrup in corn fuel ethanol fermentation production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation and Properties of Cellulose Exoglucanase CBH I Mutant

[0029] 1. Sequence of the mutant

[0030] The amino acid sequence of Trichoderma reesei cellulose exoglucanase CBH I is shown in SEQ ID NO:1, and the gene cDNA sequence is shown in SEQ ID NO:2.

[0031] After analysis by the molecular simulation software PROPKA software, the tyrosine (Y) at position 68 of the amino acid sequence of Trichoderma reesei cellulosic exoglucanase CBH I was replaced with glycine (G) to obtain mutant Y68G, and the corresponding replacement Codon (substitute GGT for TAC at positions 202-204 in SEQ ID NO:2). For Escherichia coli, the website http: / / www.jcat.de was used for codon optimization, and finally the coding gene of mutant Y68G (SEQ ID NO: 3) was obtained. In addition, the tyrosine (Y) at position 68 of the amino acid sequence of Trichoderma reesei cellulose exoglucanase CBH I was replaced with alanine (A) to obtain mutant Y68A, and the mutation was obtained by...

Embodiment 2

[0044] The preparation of embodiment 2 viscosity reducer

[0045] Viscosity reducers of the following compositions were prepared respectively. The parts in this embodiment are parts by mass. The original cellulosic exoglucanase CBH I in this example, two kinds of mutants Y68G and Y68A, respectively obtain the crude enzyme liquid according to the method in Example 1, then according to Document 1 (Pan Feng et al., Usami Aspergillus Y- 26 The purification and enzymatic properties of cellulase, the method in Nanjing University of Science and Technology, 2001) method to purify each cellulosic exoglucanase and freeze-drying, the specific activity of cellulosic exoglucanase CBH I is 21500U / g, the specific activity of mutant Y68G is 39600U / g, and the specific activity of Y68A is 38400U / g. The information of the remaining enzymes used is shown in Table 2.

[0046] Enzyme information used in the compound viscosity reducer of table 2

[0047] Enzyme name

factory

pro...

Embodiment 3

[0054] Application of embodiment 3 viscosity reducer

[0055] The technological process of corn fuel ethanol fermentation is: corn flour—pulp mixing—adding enzyme preparation and yeast—raw meal fermentation—distillation—dehydration—fuel ethanol. The addition amount of each substance in the fermentation is 2kg of corn flour, 0.1-0.5% of yeast, 20-80U / g corn flour of alpha-amylase, and 100-200U / g corn flour of glucoamylase. Fermentation conditions: material-to-water ratio 1:2-1:5, constant temperature 28-35°C, stirring speed 100-200r / min, fermentation without cooking for 3-6 days. After the fermentation is finished, the fuel ethanol is distilled out, and the distillation waste mash is filtered to obtain a clear liquid. Concentrate the supernatant to a syrup with a concentration (mass percentage of dry matter, the same below) of 29%. Viscosity reduction agent 1 and comparison viscosity reduction agent 1, viscosity reduction agent 4 and comparison viscosity reduction agent 2 in ...

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 provides a mutant of cellulose exocellobiohydrolase CBH I and viscosity reducer of syrup in corn fuel ethanol fermentation production and relates to the field of biotechnology. The mutant of the cellulose exocellobiohydrolase CBH I is obtained by replacing the 68th tyrosine of the amino acid sequence of the cellulose exocellobiohydrolase CBH I with glycine or alanine. The viscosity reducer is a mixture of the mutant of the cellulose exocellobiohydrolase CBH I and one or more than two of pectinase, xylanase, beta-glucanase, mannase, glutamine transaminase and lipase. The mutant of the cellulose exocellobiohydrolase CBH I is great in enzyme stability and catalytic activity, and the preparation method is simple and efficient. By adopting the viscosity reducer, viscosity reduction can be conducted on the syrup in the corn fuel ethanol fermentation production rapidly and efficiently under the condition of low energy consumption.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a mutant of cellulose exoglucanase CBH I and its application in reducing the viscosity of the clear liquid concentrate in starch fermentation ethanol. Background technique [0002] For a long time, fossil energy has been one of the main energy sources that human beings rely on. However, fossil energy is a non-renewable resource, which is currently being exhausted after being exploited and used in large quantities for a long time; in addition, the long-term use of fossil resources has also brought about increasingly serious environmental pollution problems. In view of the above reasons, countries around the world have been actively looking for various renewable and clean energy sources that can replace fossil resources, among which fuel ethanol is one of the most popular renewable clean energy sources. [0003] However, since 2014, due to the 60% drop in international crude oil price...

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): C12N9/42C12N9/46C12N9/10C12N9/20C12N9/88C12N15/56C12N15/70C12P7/10
CPCC12N9/1096C12N9/20C12N9/2437C12N9/2454C12N9/88C12P7/10Y02E50/10
Inventor 黄和俞亚东李秀娟李干禄
Owner NANJING TIANKAI BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com