Method for extracting and isolating clostridium acetobutylicum exopolysaccharide

A technology of Clostridium acetobutylicum and extracellular polysaccharide, which is applied in the field of bioengineering, can solve the problems that the research on the biofilm of Clostridium acetobutylicum is still in its infancy and has little research, and achieves the effects of low cost and simple operation.

Active Publication Date: 2016-10-12
NANJING UNIV OF TECH
View PDF6 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Clostridium acetobutylicum secretes a large amount of exopolysaccharides during immobilized fermentation to produce butanol. However, there are few studies on the biofilm of Clostridium acetobutylicum at this stage. The structure of Clostridium acetobutylicum biofilm and The composition research is still in it

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 extracting and isolating clostridium acetobutylicum exopolysaccharide
  • Method for extracting and isolating clostridium acetobutylicum exopolysaccharide
  • Method for extracting and isolating clostridium acetobutylicum exopolysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Inoculate 200ml of the cultured Clostridium acetobutylicum (Clostridium acetobutylicum ATCC 824) seed liquid into an anaerobic fermenter equipped with 1.8LP2 fermentation broth, and place about 60 grams of fixed The immobilized material was cultured at a constant temperature of 37°C, and the culture was repeated for 12 batches, and each batch was cultivated for 12 hours. A thick layer of Clostridium acetobutylicum biofilm would grow on the surface of the immobilized material.

[0031] The cultivated biofilm was washed twice with ultrapure water to remove free bacteria and residual fermentation broth on the surface of the biofilm. The cultured biofilm was collected and centrifuged at a high speed (10000rpm, 10min, 4°C) to obtain a biofilm with a volume of about 50ml.

Embodiment 2

[0033] Take 12 5ml centrifuge tubes, labeled 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, add 1ml of biofilm respectively, then add 0.075mol / L, 0.1mol / L, 0.125mol / L three concentrations of NaOH solution, Na 2 CO 3 Each 1ml of solution and EDTA solution was shaken for 1h, and the dissolution situation was observed (dissolution rate=10000rpm centrifugation after precipitation volume / volume of initial biofilm), as shown in the following table:

[0034] Table 1 Dissolution of biofilm under different concentrations of different solutions

[0035]

NaOH solution

Na 2 CO 3 the solution

NaHCO 3 the solution

ammonia

0.075mol / L

70％

40％

20％

50％

0.1mol / L

100％

45％

30％

60％

0.125mol / L

100％

45％

35％

65％

[0036] As shown in Table 1, the effect of dissolving biofilm with 0.1 mol / L NaOH is the best, and the dissolution rate reaches 100%, so 0.1 mol / L NaOH is finally chosen to dissolve biofilm.

[0037] Ta...

Embodiment 3

[0039] Take a 50ml centrifuge tube, add 10ml of biofilm, then add 20ml of 0.1mol / L NaOH solution, oscillate to completely dissolve the biofilm, and centrifuge at high speed to remove the sediment of the lower layer of bacteria, and collect about 28ml of supernatant 1.

[0040] Take nine 10ml centrifuge tubes, draw about 2ml of the above supernatant 1, slowly add 1.6ml, 3ml, 6ml of absolute ethanol, acetone and butanol respectively, shake evenly, the solution becomes turbid and precipitates, and centrifuge at a high speed (10000rpm , 10min, 4°C) to obtain a precipitate, that is, exopolysaccharide A, freeze-dry and measure the dry weight, as shown in the following table.

[0041] The dry weight of the polysaccharide precipitated under different concentrations of different solvents in table 2

[0042]

ethanol

acetone

Butanol

45％

14.3mg

8.5mg

7.2mg

60％

21.6mg

9.6mg

8.3mg

75％

21.5mg

10.3mg

9.8mg

[0...

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
Relative molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention provides a method for extracting and isolating clostridium acetobutylicum exopolysaccharide, comprising the following steps: (1), inoculating clostridium acetobutylicum seed liquid to a fermentation tank, adding mobilizing material, culturing until the surface of the mobilizing material is fully covered by clostridium acetobutylicum biofilm; (2), dissolving clostridium acetobutylicum biofilm with alkaline solution, centrifuging to remove bacterial precipitate, adding an organic solvent into supernate so that the organic solvent is 30-90% by weight, the organic solvent being ethanol, acetone or butanol, centrifuging, and collecting precipitate; adding an acid to adjust pH to 3.5-10.0, and collecting precipitate, the precipitates collected twice being the exopolysaccharide. The method provided herein has the advantages of low cost and good operation simplicity.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and specifically designs a method for extracting and separating the exopolysaccharide of Clostridium acetobutylicum. Background technique [0002] Studies have found that when Clostridium acetobutylicum contacts the immobilization medium, the cells aggregate and secrete autopolymers to wrap themselves, forming a special cell population structure on the surface of the medium—biofilm. Microorganisms in biofilms live in a self-produced matrix called extracellular polymers (EPS), the main components of which are polysaccharides and proteins. [0003] Polysaccharides refer to carbohydrate compounds composed of more than 20 monosaccharides. They are one of the important biomacromolecules in living organisms. , anti-aging, hypolipidemic, hypoglycemic, anti-inflammatory, anti-viral and other physiological functions, and has gradually become the research and development focus of modern biomedicine,...

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): C08B37/00
CPCC08B37/0003C08B37/006
Inventor 应汉杰王岩岩柳东沈晓宁许佳慧庄伟
Owner NANJING UNIV OF TECH
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