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Method for extracting high purity PHAs (Polyhydroxyalkanoates) from wet thallus

A polyhydroxyalkanoate, high-purity technology, used in the field of bioengineering, can solve the problems of long extraction time of acetone, long extraction time, increase of process procedures, etc., to reduce investment in production equipment, short extraction and stirring time, high molecular purity high effect

Inactive Publication Date: 2006-07-12
深圳市奥贝尔科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of the aqueous phase extraction method (non-organic solvent extraction method) are: 1. the molecular structure, especially the molecular weight, is greatly damaged; 2. the total cost of extraction is high, and the waste water treatment is troublesome; It is difficult to achieve a purity of more than 97%. There are always a small amount of impurities such as inorganic salts or proteins. These impurities have a considerable impact on the performance of the product in the post-processing, especially for film-forming products. strength, and will accelerate molecular weight degradation under processing conditions
The disadvantage of this method is that it does not take into account the loss of its high boiling point non-solvent in the biological residue (after extracting the polymer)
If it is not recovered, obviously the loss of non-solvent will increase the total solvent usage, thereby increasing the total cost; if it is recovered, due to its high boiling point, the recovery cost is high, making the total production cost higher
At the same time, this method will form small particles similar to gel in the process of removing the solvent. Such small gel particles will "enclose" the impurities (soluble in non-solvent) in the system. After filtering and drying, these "enclosed" "Impurities are difficult to remove
Therefore, the product obtained by this method has color, and its purity is difficult to reach more than 99%, and it is difficult to be suitable for products similar to medical and food packaging.
Aisao. Nuo Da proposed " one-step extraction method of oil and PHA and subsequent solvent elution ", " oil and PHA One-step extraction method and subsequent non-solvent-induced precipitation", "two-step continuous extraction method of oil and PHA", the disadvantages of these methods are: ① they are all aimed at plants or oily biomass, at this stage, the PHA content in plants All are very low, and the highest content is reported to be about 14% (by dry weight) in the leaves of Arabidopsis poly(3HB), which is not suitable for microbial fermentation broth, and the extraction yield is low with this method; ② acetone extraction The time is long, and the efficiency of wet bacteria is low. Generally, wet bacteria contain at least 50% water. If it is dried, the process procedure will be increased and the cost of extraction will be increased; ③ too much organic solvent will be lost in the biological residue
The disadvantages of this method are: 1. the wall-breaking extraction takes a long time, requiring 1-10 hours, and the efficiency is low; 2. the use of five- and / or four-carbon alcohols makes recovery difficult, solvent loss increases, and the production cost is relatively high; 3. the obtained product has low purity (less than 96%); ④ PHAs extraction yield is low, and need to carry out pretreatment, increased process procedure; ⑤ too many organic solvents lost in the biological residue

Method used

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  • Method for extracting high purity PHAs (Polyhydroxyalkanoates) from wet thallus
  • Method for extracting high purity PHAs (Polyhydroxyalkanoates) from wet thallus
  • Method for extracting high purity PHAs (Polyhydroxyalkanoates) from wet thallus

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Effect test

Embodiment 1

[0069] Extraction object: 2000ml fermentation broth containing P3HB4HB (15% 4HB) in cells

[0070] Cell concentration: 150 g (dry weight) / liter of fermentation broth

[0071] P3HB4HB content: 80% of dry cell weight

[0072] pH value: 7

[0073] The specific steps of this embodiment are as follows:

[0074] Take 2000ml of fermentation broth, centrifuge with a low-speed centrifuge (LXJ-IIB, Shanghai Anting Scientific Instrument Factory) to remove the supernatant, the cell content is 30%, to obtain 1000ml of fluid cell, add 6000ml of ethyl acetate , Stir for 25 minutes, separate the solution, collect the organic phase, this organic phase is the ethyl acetate solution with P3HB4HB dissolved; the once extracted bacteria are extracted twice according to the above operation steps, each time adding 3000ml ethyl acetate Ester, the organic phase separated for the third time is used as the solvent for the next fresh bacteria. Put the organic phase obtained from the previous two separations...

Embodiment 2

[0076] Extraction object: 2000ml fermentation broth containing P3HB4HB (15% 4HB) in cells

[0077] Cell concentration: 150 g (dry weight) / liter of fermentation broth

[0078] P3HB4HB content: 80% of dry cell weight

[0079] pH value: 7

[0080] The specific steps of this embodiment are as follows:

[0081] Take 2000ml of fermentation broth and centrifuge with a low-speed centrifuge (LXJ-IIB, Shanghai Anting Scientific Instrument Factory) to remove the supernatant, and the bacterial content is 30%. Get 1000ml of fluid bacteria, add 6000ml of ethyl acetate, stir for 30 minutes, separate the solution, collect the organic phase, this organic phase is the ethyl acetate solution with P3HB4HB dissolved; the once extracted bacteria according to the above The operation steps were repeated twice more, adding 3000 ml of ethyl acetate each time, and the organic phase separated for the third time was used as the solvent for the next fresh bacteria. Pour the organic phase obtained from the pr...

Embodiment 3

[0083] Extraction object: 100L of fermentation broth containing P3HB4HB (15% 4HB) in the cell

[0084] Cell concentration: 150 g (dry weight) / liter of fermentation broth

[0085] P3HB4HB content: 80% of dry cell weight

[0086] pH value: 7

[0087] The specific steps of this embodiment are as follows:

[0088] Take 100L of fermentation broth, centrifuge with a disc centrifuge (DHC / DRY-250, Liaoyang Sunshine Pharmaceutical Machinery Co., Ltd.) to collect the bacteria, and the bacteria content is 25%. Obtain 60L of fluid cell, add 210L of ethyl butyrate, stir for 25 minutes, separate the solution, collect the organic phase, this organic phase is the ethyl acetate solution with P3HB4HB dissolved; press the once extracted cell The above-mentioned operation steps were repeated for two more extractions, each adding 70L of ethyl butyrate, and the organic phase separated for the third time was used as the solvent for the next fresh bacterial cells. Pour the organic phase solution obtain...

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Abstract

The invention relates to a method for extracting high purity PHAs (polyhydroxy etianate) from the humid thalline, which comprises a method for directly extracting polyhydroxy etianate from humid biology and a method for obtaining a high purity polyhydroxy etianate. It comprises the following steps: compressing the fermentation liquor, collecting the thalline, adding organic solution into the humid thalline to dissolve the macro mole, separating the residuum, reserving the organic phase with PHAs. It compresses the organic phase with PHAs to obtain PHAs product; it uses non-solvent to treat the organic phase with PHAs to obtain the deposition which is PHAs. The obtained PHAs uses polarity non-solvent to treat, filter and dry to obtain high purity PHAs product.

Description

Technical field [0001] The invention belongs to the field of bioengineering technology and relates to a method for directly extracting polyhydroxyalkanoates from biological systems such as bacteria using organic solvents. It mainly refers to a method for extracting high-purity polyhydroxyalkanoates from wet bacterial cells: wet extraction method. Background technique [0002] Polyhydroxyalkanoates (PHAs) are usually granular substances accumulated in cells by microorganisms under unbalanced growth conditions for storing carbon sources and energy. They have physical properties similar to polyethylene, polypropylene, etc. Thermoplastic polyester is a chemically synthesized plastic, but it has characteristics that general synthetic plastics do not have: biodegradability, biocompatibility, piezoelectricity, optical activity, etc., and is considered a "new biodegradable material". Much attention is paid to it. [0003] Fermentation technology for PHAs has developed greatly due ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G63/06C12P19/00
Inventor 夏帮富
Owner 深圳市奥贝尔科技有限公司