Double-bacteria system construction method for PHA accumulation

A construction method and system technology, applied in the field of mixed flora research in microbiology, can solve the problems that xylose cannot be rapidly utilized by microorganisms, heavy metabolic burden of strains, etc., and achieve the effect of avoiding excessive metabolic load and high utilization efficiency

Active Publication Date: 2017-05-10
TIANJIN UNIV
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Problems solved by technology

The reason is that at present, PHA is mainly produced by the traditional fermentation of a single microbial strain [4], xylose itself cannot be rapidly utilized by microorganisms [5], and it is expe

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  • Double-bacteria system construction method for PHA accumulation
  • Double-bacteria system construction method for PHA accumulation
  • Double-bacteria system construction method for PHA accumulation

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[0041] The following describes the present invention in further detail with reference to the drawings and specific embodiments:

[0042] Saccharomyces cerevisiae is a gift from Professor Yuan Yingjin of Tianjin University, and its starting strain is Saccharomyces cerevisiae BY 4741; PHA-accumulating Pseudomonas putida NBRC 14164, purchased from the China Industrial Microbial Culture Collection and Management Center. The design of the two bacteria relationship is like figure 1 Shown.

[0043] 1. Medium and solution formulation

[0044] YPX medium: peptone 20g / L, yeast powder 10g / L, xylose 20g / L;

[0045] LB medium: peptone 10g / L, yeast powder 5g / L, sodium chloride 10g / L;

[0046] Inorganic salt culture medium: ammonium sulfate 3g / L, magnesium sulfate 0.5g / L, dipotassium hydrogen phosphate 3g / L, potassium dihydrogen phosphate 1.5g / L, carbon source 20g / L, trace element 1mL / L;

[0047] Trace elements: ferrous sulfate 2.78g / L, calcium chloride 1.47g / L, manganese chloride 1.98g / L, cobalt chlo...

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Abstract

The invention discloses a double-bacteria system construction method. Brewer's yeast and pseudomonas putida are adopted to form a double-bacteria system I, the brewer's yeast realizes xylose utilization, and the pseudomonas putida realizes PHA accumulation. Studies on the system mainly include: determining lactic acid as an optimal carbon source of the pseudomonas putida, and on the basis, designing a middle exchange metabolism product of double bacteria to be lactic acid; exploring an optimal inoculation proportion of the double bacteria, and setting a standard of inoculating 2 g of pseudomonas putida liquid into each 200mL of inorganic salt-xylose culture medium, wherein when the inoculation proportion of the brewer's yeast and the pseudomonas putida is 10:1, talli grow most fast and are highest in biomass; exploring adding time point of sodium caprylate which is a PHA precursor substance, wherein when adding amount is 5g/L and the adding time point is at the beginning of fermentation, PHA accumulation reaches a highest level.

Description

technical field [0001] The invention relates to a method for constructing a dual-bacteria system for PHA accumulation, which belongs to the research field of mixed flora in microbiology. Background technique [0002] Polyhydroxyalkanoate (PHA) has attracted widespread attention as an excellent substitute for traditional plastics due to its good biocompatibility, environmental friendliness, and renewability of raw materials [1], and it is also widely used in the field of biomedicine. widely used. However, the cost of PHA produced by existing methods is much higher than that of traditional plastics, which limits its large-scale production and application [1]. According to a survey by Tsinghua University, substrate consumption accounts for 50% of the cost of PHA. In order to reduce substrate costs, people have turned their attention from expensive substrates such as glucose to non-grain raw materials, such as xylose, which accounts for 18-30% of natural lignocellulose content....

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Application Information

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IPC IPC(8): C12P39/00C12P7/62C12R1/865C12R1/40
CPCC12P7/625C12P39/00
Inventor 贾晓强刘畅
Owner TIANJIN UNIV
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