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

A method for the production of bio-based chemicals using highly cellulose hydrolyzate tolerant Escherichia coli

A technology of cellulose hydrolyzate and Escherichia coli, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problems of difficult operation of foreign plasmids, unstable and easy degradation of host strains, and achieve Mild conditions, easy genetic engineering, and environmentally friendly effects

Active Publication Date: 2015-11-04
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such engineering strains can only target a certain toxic compound, and cannot effectively solve the problem of a wide variety of toxic compounds (furfural, hydroxymethylfurfural, acetic acid, phenolic compounds) in the lignin enoic acid hydrolyzate
[0004] Although the genetic background of Escherichia coli is relatively clear, the low-cost lignocellulose hydrolyzate in the production industry of existing bio-based chemicals contains toxic substances that can inhibit engineered Escherichia coli, and the host strains obtained through domestication are unstable and easy Degradation, the host bacteria obtained through genetic engineering and containing foreign plasmids are not easy to operate, and it is a feasible method to obtain a strain resistant to cellulose dilute acid hydrolyzate by mutagenesis

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 method for the production of bio-based chemicals using highly cellulose hydrolyzate tolerant Escherichia coli
  • A method for the production of bio-based chemicals using highly cellulose hydrolyzate tolerant Escherichia coli
  • A method for the production of bio-based chemicals using highly cellulose hydrolyzate tolerant Escherichia coli

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: strain mutagenesis and screening

[0026] Measure the number of cells in the suspension by hemocytometer, and dilute to 105-106 cells / mL with sterile water. Take 5mL of bacterial suspension in a 9cm-diameter plate, put it into a rotor sterilized by 70% ethanol, and stir it magnetically. Put it under a 30w ultraviolet lamp at a distance of about 25cm. After a certain period of irradiation (1s, 2s, 5s, 15s, 30s, 60s), take 0.1mL and spread it on a lignocellulose hydrolyzate selective medium plate and culture it in the dark at 37°C After 16-18 hours, count the number of colonies on the plate, calculate the lethal rate, and draw the lethal curve.

[0027] The lignocellulose hydrolyzate culture medium is obtained by the following method: add 450mL distilled water and the following components in a 1 L beaker to dissolve it in water: NH 4 Cl 0.3~0.5g, Na 2 HPO 4 2~3g, KH 2 PO 4 1 ~ 2g, adjust the pH to 7.4 with 1 M NaOH and adjust the total volume to 450 ~ 550...

Embodiment 2

[0033] Example 2: Verification of strain tolerance to lignocellulose hydrolyzate

[0034] Preparation of lignocellulose alkenoic acid hydrolyzate: take the dried stalks, filter the crushed corn stalks with 40 mesh according to the solid-liquid ratio of 5%, hydrolyze them in 1% sulfuric acid at 100°C for 5 hours, filter and adjust the pH It is later used as a carbon source for cellulose hydrolysis, and its main components are glucose, xylose, and furfural, and also contains a small amount of hydroxymethylfurfural, acetic acid, phenolic compounds, and sodium sulfate produced by neutralization with sodium hydroxide.

[0035] Pick the strains resistant to lignocellulose hydrolyzate obtained by screening, use the strains that have not undergone mutagenesis, and inoculate them in a test tube containing 2 mL of LB medium, and inoculate when the cell density is about 1 In a 250mL shake flask equipped with 50mL of the above-mentioned lignocellulose hydrolyzate 3% as the sole carbon sou...

Embodiment 3

[0038] Example 3: Synthesis of bio-based chemicals using cellulose hydrolyzate tolerant host bacteria

[0039] Competent strain preparation:

[0040] 1. Pick a single colony with a diameter of about 2 to 3 mm from an overnight (16-20h) culture plate at 37°C. Such a single colony was inoculated into a sterilized 30 ml test tube containing 5 ml of LB broth, and cultured overnight at 37°C with shaking.

[0041] 2. Transfer 0.2 ml of overnight culture to a 50 ml sterilized Erlenmeyer flask containing 15 or 20 ml of LB. Shake culture at 37°C for 2 to 2.5 hours (the bacteria are in the logarithmic growth phase at this time).

[0042] 3. Centrifuge at 4000rpm for 5 minutes at room temperature to collect the logarithmic phase cells. Discard the medium and keep the cell pellet.

[0043] 4. Add 10 mL of ice-cold MgCl 2 -CaCl 2 solution, and beat lightly.

[0044] Cells were collected by centrifugation at 4000 rpm for 10 minutes at 5.4°C.

[0045] 6. Discard MgCl 2 -CaCl 2 solut...

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 producing a bio-based chemical by adopting high-cellulose hydrolysate tolerant escherichia coli, and particularly provides a host escherichia coli strain, which can be used for producing the bio-based chemical, of tolerant lignocellulose hydrolysate. The host escherichia coli strain provided by the invention has very good lignocellulose hydrolysate tolerance, can efficiently and directly utilize the lignocellulose hydrolysate and can be used for producing the bio-based chemical through metabolic engineering reform.

Description

technical field [0001] The invention relates to an Escherichia coli with high tolerance to cellulose hydrolyzate and a method for fermenting and producing bio-based chemicals. Background technique [0002] With the development of biochemical technology, a large number of metabolic pathways of bio-based chemicals have been constructed in Escherichia coli hosts to achieve the biosynthesis of the chemicals, and because of the high added value of bio-based chemicals, they have been favored by biochemists. favor. However, how to effectively reduce the cost of raw materials has become a bottleneck restricting the chemical industry of bio-based chemicals. The price of sugar, the main raw material in the fermentation process, becomes a major factor limiting its cost. Dilute acid hydrolysis of lignocellulose can effectively reduce the cost of fermentation raw materials, but a variety of fermentation inhibitors will be formed in the product, such as furfural, hydroxymethylfurfural, ...

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): C12N1/20C12P5/02C12P7/42C12P5/00C12R1/19
Inventor 咸漠张海波赵国明刘玉华
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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