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Method for improving yield of PHB produced by Ralstonia eutropha by optical driving

An Alcaligenes eutropha, light-driven technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as insufficient understanding of microbial nutritional needs, increased production costs, and low success probability. Achieve the effect of promoting growth and production capacity, increasing yield, and low cost

Active Publication Date: 2018-12-21
WUHAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods of improving microbial metabolites are mainly manifested in three aspects. First, the selection and optimization of the medium is often subjective and arbitrary, and there is no way to know what kind of metabolites will be produced by a certain bacterium. Second, the use of nutrient-rich media will not only increase production costs, but also be limited to a certain extent by the lack of comprehensive understanding of microbial nutritional requirements; third, by introducing or changing specific genomes, this method requires Very professional personnel design and countless repeated experiments, the process is complicated and the probability of success is low

Method used

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  • Method for improving yield of PHB produced by Ralstonia eutropha by optical driving
  • Method for improving yield of PHB produced by Ralstonia eutropha by optical driving
  • Method for improving yield of PHB produced by Ralstonia eutropha by optical driving

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Utilize CdS to improve the amount of poly-beta-hydroxybutyrate (PHB) produced by Ralstonia eutropha (Bei Na Biology, BNCC137386), the steps are as follows:

[0032] (1) Preparation of CdS

[0033] Prepare 35 mL each of 0.2 mol / L cadmium nitrate and 0.2 mol / L sodium sulfide solution, add the sodium sulfide solution dropwise into the cadmium nitrate solution under stirring conditions, and after stirring for 2 hours, put the mixed solution into polytetrafluoroethylene In an ethylene reactor, gradually raise the temperature to 200°C and keep it warm for 6 hours. After the reaction was completed, it was naturally cooled to room temperature, and the obtained precipitate was suction-filtered, washed, and dried at 55° C. to obtain a yellow CdS powder.

[0034] (2) Microbial culture

[0035] The strains stored in the freeze-drying tube were activated in the seed medium. After Ralstonia eutropha grew for 24 hours, the obtained bacterial solution was inserted into the basic medi...

Embodiment 2

[0040] Using CdS / Cd(OH) 2 To increase the amount of poly-β-hydroxybutyrate (PHB) produced by R.eutropha, the steps are as follows:

[0041] (1) CdS / Cd(OH) 2 Preparation of Composite Photocatalyst

[0042] Weigh 8.3291g NaOH dissolved in 40mL deionized water, measure 10mL triethanolamine dissolved in NaOH solution, weigh 0.4574g CdCl 2 2.5H 2 O was dissolved in 10mL deionized water, and then weighed 0.4926g Na 2 S·2H 2 O dissolved in 10 mL deionized water, CdCl 2 Add the solution to the mixed solution of triethanolamine and NaOH, stir evenly, then add NaOH dropwise to the mixed solution 2 After stirring the S solution evenly, put the mixed solution into a polytetrafluoroethylene reactor, gradually raise the temperature to 180°C, and keep the temperature for 12h. After the reaction, cool down to room temperature naturally, filter the obtained precipitate, wash it, and dry it at 55°C to obtain CdS / Cd(OH) 2 composite photocatalyst.

[0043] (2) Microbial culture

[0044]...

Embodiment 3

[0049] Use g-C 3 N 4 To increase the amount of poly-β-hydroxybutyrate (PHB) produced by Ralstonia eutropha, the steps are as follows:

[0050] (1) g-C 3 N 4 preparation of

[0051] Weigh 10g of urea, dry it at 55°C, move it to a muffle furnace for 2 hours at 550°C, and cool it down to room temperature naturally after the reaction, filter and wash the obtained precipitate, and dry it at 55°C to obtain Yellow g-C 3 N 4 powder.

[0052] (2) Microbial culture

[0053] The strains stored in the freeze-drying tube were activated in the seed medium. After Ralstonia eutropha grew for 24 hours, the obtained bacterial solution was inserted into the basic medium I for expansion and cultivation until the od value was 1, and the same amount of bacterial solution was transferred to To two groups of basal medium, one of which added g-C 3 N 4 (Concentration is 1g / L), another group does not add anything, cultures in Erlenmeyer flask at 30°C, 200 rpm for 84h, the medium volume of each...

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Abstract

The invention relates to a method for improving yield of PHB produced by Ralstonia eutropha by optical driving. The method comprises the following specific steps: 1) microbial culture: activating theRalstonia eutropha stored in a freeze-drying tube in a seed culture medium, and after the Ralstonia eutropha grows for 24h, inoculating the obtained bacterial solution into a basal culture medium to perform enlarging cultivation until the od value is 1; 2) passaging the Ralstonia eutropha subjected to enlarging cultivation to a basal culture medium II containing a photocatalyst and an electron donor; 3) collecting the bacterial solution in the basal culture medium II and extracting PHB. The method provided by the invention only needs to directly add an appropriate amount of preferred kind of photocatalyst and electron donor to a suitable microbial culture medium, and can induce the microorganism to generate more metabolites. The method has the advantages of simple operation, low cost, mildcondition, environmental friendliness and the like.

Description

technical field [0001] The invention relates to a method for increasing the yield of PHB produced by Ralstonia eutropha by light-driven. Background technique [0002] Poly-β-hydroxybutyrate (PHB) is a biodegradable and renewable bioplastic that can be synthesized by a variety of organisms, stored in cells, and can be re-decomposed for cell growth when carbon sources are scarce. carbon and energy sources. Although PHB is considered an effective alternative to conventional plastics in medical, agricultural and food packaging, full-scale commercialization of this biopolymer has been hampered by its high production cost. In order to overcome these limitations, the production of PHB by bacterial culture has aroused extensive interest of researchers. Ralstonia eutropha H16 is an excellent strain capable of accumulating PHB. It can synthesize PHB in a large amount using fructose, which can reach 80% of the dry cell weight (CDW). Ralstonia eutropha is a ubiquitous soil and freshw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62C12R1/01
CPCC12P7/625
Inventor 张甜许梦莹皮埃尔鲁克·川柏雷胡胜军
Owner WUHAN UNIV OF TECH
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