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A dynamic control system for blue light control and its application

A blue light and photosensitive 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, etc., and achieve the effect of simple design, less system components, and low strain growth load

Active Publication Date: 2021-10-29
JIANGNAN UNIV
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  • Summary
  • 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 uneven mass transfer of dissolved oxygen, which directly affects the growth and accumulation of acetoin in E. coli

Method used

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  • A dynamic control system for blue light control and its application
  • A dynamic control system for blue light control and its application
  • A dynamic control system for blue light control and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Evaluation of Blu-ray activation elements under different light intensity and optical pulse cycle

[0047] Put P Trc -E and PBLIND-MKATE plasmid transfer to E.Coli F0601 (see: dong x, chen x, qian y, et al.metabolic engineering of escherichia coli w3110 to production l-malate [J] .biotechnology & BioEngineering, 2016, In 114 (3): 656-664. In the engineering strain, it was cultured in the LB medium, and the SPECTramax M3 microplate laminator was used to continuously fluorescence measurement. When strains are in 450 nm Blu-ray conditions, the Blu-ray photoreactive protein EL222 is combined to P lux Or P J23119 The expression of fluorescent protein mkate is achieved before the promoter.

[0048] Experiments under dark or 450 nm Blu-ray light, figure 1 As shown, a figure shows the dose dependence of the lighting strength of the blue-ray system, with the increase of blue light light intensity, increased from 242.68OD (Au) from 242.68OD (Au); B figure shows that Blu-ray...

Embodiment 2

[0049] Example 2 Dynamic gene line introduced a shake flask fermentation performance of the production strain of B

[0050] like image 3 As shown in P Trc -E-P J23119 -Budab-Nox and PBLIND-FTSZ-FTSA (granular map figure 2 In order to express the carrier, different dynamic gene lines were introduced into the E.Coli F0601 strain, and 7 strains of engineered strains such as D1-D7 were obtained, and the D1-D2 strain represented only P. Trc -E-P J23119 -Budab-Nox, PBLIND-FTSZ-FTSA, D3-D7 engineered strain is commonly expressed Trc -E-P J23119 -Budab-Nox and PBLIND-FTSZ-FTSA, D3-D7 respectively indicate that the blue light light intensity is 0.2 w / cm, respectively. 2 0.25W / cm 2 0.3W / cm 2 0.4W / cm 2 , The engineering strain at 0.8w / cm.

[0051] like Figure 4 with 5 As shown, the growth and product synthesis of strains in the NBS inorganic salt medium are detected. The shake flask fermentation results show that the ethlancer production of D1-D2 strains increased by 19.5 g / L fro...

Embodiment 3

[0052] Example 3 Dynamic gene line introduced the fermentation tank fermentation performance of the production strain of B

[0053] The fermentation properties of the D7 strain were detected in a 5L fermentor. The temperature is constant 37 ° C, 500 rpm, the initial vaccination amount is controlled to 5%, the ventilation is 1 Vvm, the fermentation period is 72 h. At the end of the fermentation, the accumulation of B.

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Abstract

The invention discloses a blue-light-controlled dynamic regulation system and its application, specifically discloses a blue-light-controlled dynamic regulation system and its application in the production of acetoin, belonging to the technical field of bioengineering. The present invention selects the blue light photosensitive protein by means of molecular biology, utilizes the light-sensitive protein’s specific response to the light source characteristic of a specific wavelength and the advantage of not needing to add an inducer from an external source, and introduces the dynamic regulation of targeting FtsZ+FtsA into engineering Escherichia coli The gene circuit realizes the production of acetoin in the inorganic salt medium. The yield of acetoin produced by the invention can reach 63g / L, and the conversion rate can reach 0.57g / g glucose.

Description

Technical field [0001] The present invention relates to a dynamic regulatory system of blue-ray control and its application, and there is a dynamic regulatory system of blue-ray control and its application in the group of B, which belongs to the field of bioengineering. Background technique [0002] The dynamic regulatory system is an emerging metabolic stream regulation means in the field of metabolic engineering. The main feature of the static regulation is that the engineering strain will be made according to the fermentation time, physiological state, endydaymame concentration, and extracellular environment changes. Out of the corresponding enzyme is adjusted, which in turn affects metabolic flow distribution and improves product production capacity. The dynamic regulatory system has a fermentation process without human regulation, and does not require an advantage of exogenous addition inducers, which has a significant advantage in the production of high additional value com...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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