Dynamic control system for blue light control and application thereof

A blue light and promoter technology, applied in the field of bioengineering, can solve the problems of uneven mass transfer of dissolved oxygen, affecting the growth and accumulation of acetoin in Escherichia coli, and achieve a simple design, few system components, and low strain growth load. Effect

Active Publication Date: 2019-11-12
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the production of acetoin is limited by the specific surface area, causing the problem of unev...

Method used

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  • Dynamic control system for blue light control and application thereof
  • Dynamic control system for blue light control and application thereof
  • Dynamic control system for blue light control and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Evaluation of blue light-activated components under different light intensities and light pulse periods

[0047] Will P Trc -E and pBLind-mKate plasmids were transferred into E.coli F0601 (see: Dong X, Chen X, Qian Y, et al. Metabolic engineering of Escherichia coli W3110 to produce L-malate[J]. Biotechnology&Bioengineering, 2016, 114 (3): 656-664.), obtain engineering strain, place it in LB culture medium, use SpectraMax M3 microplate microplate reader to carry out continuous fluorescence measurement to above-mentioned engineering strain. When the strain is exposed to 450nm blue light, the blue light photosensitive protein EL222 binds to P lux or P J23119 Before the promoter, the expression of fluorescent protein mKate is realized.

[0048] Experiments were carried out under dark or 450nm blue light conditions, and the results were as follows figure 1 As shown, Figure A shows the dose dependence of the light intensity of the blue light system. With the in...

Embodiment 2

[0049] Embodiment 2 dynamic gene circuit introduces the shake flask fermentation performance of acetoin production strain

[0050] Such as image 3 shown in P Trc -E-P J23119 -BudAB-NOX and pBLind-FtsZ-FtsA (see plasmid map figure 2 ) as an expression vector, different dynamic gene circuits were introduced into the E.coli F0601 strain, and a total of 7 engineering strains such as D1-D7 were obtained, and the D1-D2 strains expressed only the expression of P Trc -E-P J23119 -Engineering strains obtained from BudAB-NOX, pBLind-FtsZ-FtsA, D3-D7 engineering strains for the co-expression of P Trc -E-P J23119 -Engineering strains obtained from BudAB-NOX and pBLind-FtsZ-FtsA, D3-D7 respectively represent blue light intensity of 0.2W / cm 2 、0.25W / cm 2 、0.3W / cm 2 、0.4W / cm 2 , Engineering strains at 0.8W / cm.

[0051] Such as Figure 4 with 5 As shown, the strain growth and product synthesis were detected in NBS inorganic salt medium. The shake flask fermentation results show...

Embodiment 3

[0052] Example 3 Fermentation performance of fermenter for introducing acetoin production strain into dynamic gene circuit

[0053] The fermentation performance of the D7 strain was tested in a 5L fermenter. The temperature is constant at 37°C, 500rpm, the initial inoculation amount is controlled at 5%, the ventilation rate is 1vvm, and the fermentation period is 72h. At the end of the fermentation, the accumulation of acetoin reached 63g / L, and the conversion rate reached 0.57g / g glucose.

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Abstract

The invention discloses a dynamic control system for blue light control and application thereof, in particular to a dynamic control system for blue light control and application thereof in acetoin production, and belongs to the technical field of biological engineering. According to the invention, the blue light photosensitive protein is selected by molecular biology means, the characteristic thatthe photosensitive protein specifically responds to a light source with a specific wavelength and the advantage that an inducer does not need to be added externally are utilized, and the production of the acetoin in an inorganic salt culture medium is realized by introducing a dynamic regulation gene line targeting FtsZ + FtsA into engineering escherichia coli. The yield of acetoin produced by the system can be up to 63 g/L, and the conversion rate can be up to 0.57g/ g glucose.

Description

technical field [0001] The invention relates to a dynamic control system controlled by blue light and its application, in particular to a dynamic control system controlled by blue light and its application in the production of acetoin, belonging to the technical field of bioengineering. Background technique [0002] The dynamic control system is an emerging metabolic flow control method in the field of metabolic engineering. Its main feature different from the static control is that during the fermentation process, the engineered strains will adjust according to the fermentation time, physiological state, intracellular metabolite concentration, and changes in the extracellular environment. The corresponding adjustment of enzyme activity can affect the distribution of metabolic flux and improve the production capacity of products. The dynamic regulation system has the advantages of no need for artificial regulation in the fermentation process and no need for exogenous additio...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12P7/26C12R1/19
CPCC07K14/195C07K14/245C12N15/70C12P7/26
Inventor 陈修来丁强刘立明李洋刘佳罗秋玲
Owner JIANGNAN UNIV
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