Polyhydroxyalkanoate synthesis bacterium and its fermentation culturing method

A technology of polyhydroxyalkanoate and culture medium, applied in the field of polyhydroxyalkanoate synthetic bacteria and its fermentation culture, can solve the problems of high cost, unstable strain characteristics, difficult to control PHA, etc., and achieve simple synthesis conditions , excellent mechanical and processing properties, and broad application prospects

Active Publication Date: 2011-10-05
NANKAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, under the conditions of different types and proportions of carbon sources, the PHA synthetic bacteria can synthesize different PHA structures. Therefore, it is not easy to control the fermentation conditions to synthesize the expected specific structure of PHA. Moreover, many strains that utilize genetic engineering There are problems such as high cost and unstable strain characteristics

Method used

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  • Polyhydroxyalkanoate synthesis bacterium and its fermentation culturing method
  • Polyhydroxyalkanoate synthesis bacterium and its fermentation culturing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Screening and isolation of Pseudomonas lundensis PHA5

[0023] The experimental materials were selected from farmland soil in Xiqingfu Village, Tianjin.

[0024] The specific implementation steps are as follows: get 1g of remaining activated sludge, stir in 99ml of normal saline, let it stand, take the supernatant, dilute it with a 10-fold gradient with normal saline, and dilute it to 10 times respectively 3 -10 7 , take 10 3 -10 6 Spread on a solid screening medium plate, incubate at 30°C for 48 hours, observe under an ultraviolet analyzer, mark orange-red fluorescent colonies, pick the colony, streak on a solid screening medium plate for purification, and incubate at 30°C for 24 hours After 1 hour, the orange-red colonies were picked and stored in glycerol tubes. After 16S rDNA gene sequence analysis and BIOLOG identification, it was determined that the strain belonged to the genus Pseudomonas and the species lundensis, and it was named Pseudomonas lun...

Embodiment 2

[0025] Example 2 Synthesis of PHA by Fermentation of Pseudomonas lundensis PHA5

[0026] The specific steps are as follows: first, inoculate the strain into 5ml LB liquid medium, and culture it on a shaker at 30°C with a rotation speed of 180rpm. After culturing for 24 hours, inoculate it into the liquid fermentation medium according to the inoculation amount of 10%, shake the flask at 30°C Ferment for 48 hours. The fermentation broth was centrifuged, the supernatant was discarded, and the cells were collected and freeze-dried. Transfer the dried cells into an Erlenmeyer flask, add chloroform and stir thoroughly for 48 hours, remove the cell residue by filtration, concentrate the filtrate by rotary evaporation, add 40 times the volume of cold methanol to precipitate the PHA product, collect the precipitate, dry it in vacuum, and store it.

Embodiment 3

[0027] Example 3 Synthesis of PHA by Fermentation of Pseudomonas lundensis PHA5

[0028] The specific steps are as follows: first, inoculate the strain into 5ml LB liquid medium, and culture it on a shaker at 30°C with a rotation speed of 180rpm. After culturing for 24 hours, inoculate it into the liquid fermentation medium according to the inoculation amount of 10%, shake the flask at 30°C Ferment for 96 hours. The fermentation broth was centrifuged, the supernatant was discarded, and the cells were collected and freeze-dried. Transfer the dried cells into an Erlenmeyer flask, add chloroform and stir thoroughly for 48 hours, remove the cell residue by filtration, concentrate the filtrate by rotary evaporation, add 40 times the volume of cold methanol to precipitate the PHA product, collect the precipitate, dry it in vacuum, and store it.

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Abstract

The invention discloses a polyhydroxyalkanoate synthesis bacterium Pseudomonas lundensis PHA5 and a method for preparing polyhydroxyalkanoate by the culturing and fermentation of the bacterium. The bacterial strain takes glucose as a single carbon source to synthesize shortest-chain and medium-long-chain copolymerized PHA. The synthesis condition of PHA with the single carbon source avoids the requirements on the types and proportion of multifarious carbon sources, therefore simplifying the synthesis condition of the shortest-chain and medium-long-chain copolymerized PHA; the PHA which is composed of multifarious shortest-chain monomers and medium-long-chain monomers has more excellent machinery and processing performance than shortest-chain PHA and medium-long-chain PHA, and has a wider application prospect.

Description

technical field [0001] The invention relates to a polyhydroxyalkanoate synthetic bacterium Pseudomonas lundensis PHA5 and a method for fermenting and synthesizing polyhydroxyalkanoate by culturing the bacterium. Background technique [0002] Polyhydroxyalkanoate (PHA) is an intracellular substance produced by microorganisms to store energy and carbon sources when the nutritional conditions are unbalanced. It has been reported that there are many kinds of microorganisms capable of synthesizing polyhydroxyalkanoate (PHA) in nature. [0003] Polyhydroxyalkanoate (PHA) is a kind of linear saturated polyester and a kind of polymer material with good biodegradability. It not only has similar properties to chemically synthesized polymer materials, but also has some special properties. Excellent properties: ①good biocompatibility; ②plasticity; ③hydrophobicity; ④low oxygen permeability; ⑤piezoelectricity; ⑥optical activity, etc. [0004] Based on many excellent properties of polyhy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/62C12R1/38
Inventor 宋存江杨小娟郭文斌王淑芳李园刘耀辉孙肇直石晓宇
Owner NANKAI UNIV
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