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1-strain pseudomonas mendocina and uses thereof

A technology of Pseudomonas sapseudomonas, NK-01CCTCCM208005, applied in bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of slow growth rate, long fermentation period, low fermentation product yield, etc., and achieve easy processing and molding , the effect of high elasticity

Inactive Publication Date: 2008-09-24
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there are disadvantages such as complex process, slow growth rate in the initial stage of fermentation, long fermentation cycle, and relatively low yield of fermentation products.

Method used

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  • 1-strain pseudomonas mendocina and uses thereof
  • 1-strain pseudomonas mendocina and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1, using the Nile Red (Nile Red) fluorescent staining method to isolate the bacterial species Pseudomonas mendoza NK-01CCTCC M 208005 from the farmland soil of Fu Village, Xiqing District, Tianjin City:

[0027] Take 0-10cm of soil and dilute it with sterile saline to 10 1 -10 8 , apply 0.5mL of soil suspension with different dilutions on PHA synthetic bacteria isolation medium 1, and culture at 30°C for 4-5 days. The orange-red fluorescent colonies are PHA-synthesizing bacteria. Then the strains were classified and identified by traditional physiological and biochemical classification experiments, 16S rRNA gene sequence analysis and BIOLOG microbial taxonomic identification system. The bacterial strain was established as Pseudomonas mendoza, named Pseudomonas mendoza NK-01, and was deposited under the number CCTCC M208005.

Embodiment 2

[0028] Example 2, one-step fermentation to prepare medium and long-chain polyhydroxyalkanoate (fermentation under high carbon and low nitrogen conditions)

[0029] The pre-cultivation is in the L-test tube, adding 5ml of medium 2 by aseptic operation, inserting a single colony of NK-01 strain with a sterile toothpick, and incubating at 30°C and 120rpm for 12h. Inoculate 0.5 ml of the pre-culture into a 500 ml Erlenmeyer flask containing 100 ml of medium 2, and culture with shaking at 150 rpm at 30°C for 24 hours. Sterile centrifuge at 6000g for 10min at 4°C, discard the supernatant, oscillate and mix with sterile phosphate buffer in a centrifuge tube, centrifuge again at 4°C, 6000g for 12min, discard the supernatant. The bacteria without medium 2 were aseptically inserted into a 1-liter fermentation device with controllable oxygen stirring, which contained 1 liter of medium 3 (pH 7), and cultured at 30°C for 60 hours with aeration. After the fermentation, the cells were centr...

Embodiment 3

[0030] Example 3, two-step method fermentation to prepare medium and long-chain polyhydroxyalkanoate (fermentation under high carbon and nitrogen-free conditions)

[0031] The pre-cultivation is in the L-test tube, adding 5ml of medium 2 by aseptic operation, inserting a single colony of NK-01 strain with a sterile toothpick, and incubating at 30°C and 120rpm for 12h. Inoculate 0.5 ml of the pre-culture into a 500 ml Erlenmeyer flask containing 100 ml of medium 2, and culture with shaking at 150 rpm at 28°C for 24 hours. Sterile centrifuge at 6000g for 10min at 4°C, discard the supernatant, oscillate and mix with sterile phosphate buffer in a centrifuge tube, centrifuge again at 4°C, 6000g for 12min, discard the supernatant. The bacteria without medium 2 were aseptically inserted into 10 bottles of 500ml Erlenmeyer flasks containing 100ml of medium 4 (pH 7), cultured at 30°C with shaking at 150rpm. Finish fermentation after 60 hours, following steps are with embodiment 2.

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Abstract

The invention discloses pseudomonas mendocina and application thereof, and relates to pseudomonas mendocina NK-01 CCTCC M 208005, and a method for producing medium-and-long-chain-length polyhydroxyalkanoate which is the copolymer of 3-hydroxy hexanoate, 3-hydroxy octanoic acid, 3-hydroxy capric acid, Delta<7>-3-jalapinolic acid and 3-hydroxy stearic acid. In the method, Pseudomonas mendocina NK-01 CCTCC M 208005 is fermented in the culture medium of carbon source including glucose or treacle, or the mixture of glucose and treacle. The invention overwhelms the defects that long chain fatty acid needs to be added in the carbon source when long-chain-length polyhydroxyalkanoate, microbial growth is inhibited easily, the fermentation rate is low and the rate of production of fermentation products is low. By adopting the strain and the production process, a novel high elasticity biologic degradable material easy to be processed and shaped, namely the copolymer of 3-hydroxy hexanoate, 3-hydroxy octanoic acid, 3-hydroxy capric acid, Delta<7>-3-jalapinolic acid and 3-hydroxy stearic acid, can be produced. In addition, the fermentation conditions and the process enable large-scale industrial production.

Description

technical field [0001] The invention relates to a strain of Pseudomonas mendoza and its application, in particular to a strain of Pseudomonas mendoza and the production of 3-hydroxycaproic acid, 3-hydroxyoctanoic acid, 3-hydroxydecanoic acid, / Δ 7 - Process for copolymers of 3-hydroxyhexadecanoic acid and 3-hydroxyoctadecanoic acid. Background technique [0002] Polyhydroxyalkanoate (PHA) is a carbon source or energy particle accumulated in the body of microorganisms under the condition of losing nutritional balance, that is, rich in carbon and low in phosphorus, low in nitrogen, or rich in carbon and deficient in phosphorus and nitrogen. These polymer particles are PHA. It has been reported that more than 300 species of microorganisms can synthesize and accumulate more than a hundred different types of PHA. Due to the characteristics of biodegradability, biocompatibility and piezoelectricity, PHA has become a hot spot in the field of biomaterials research. [0003] PHA ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/64C12R1/38
Inventor 宋存江王淑芳郑承纲张斌陶剑郭文斌刘莉刘娜胡丹
Owner NANKAI UNIV
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