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High-dissolved oxygen bioreactor for high-density cultivation of genetically engineered bacteria and cultivation control method

A technology for high-density culture and genetically engineered bacteria, which is applied in the field of high-dissolved oxygen bioreactors, can solve the problems of unfavorable growth of shear-sensitive microorganisms, unfavorable growth of microorganisms, and weak macro-mixing strength, and achieve convenient large-scale operation and gas Good liquid dispersion performance and the effect of overcoming the lack of dissolved oxygen

Inactive Publication Date: 2014-10-08
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ventilated stirred reactor is suitable for most aerobic fermentation processes, but there are also deficiencies. For example, in order to achieve a higher dissolved oxygen concentration and mixed mass transfer, it is necessary to increase the stirring speed and aeration volume, which will increase energy consumption and make the The overall shear stress in the tank increases, which is not conducive to the growth of shear-sensitive microorganisms, especially when the filamentous bacteria are cultured, the damage to the cells is greater
Since the gas-liquid dispersion rotor is small and placed at the bottom of the tank, during the fermentation process, the fermentation liquid in the middle and upper part of the tank is only driven by the air bubbles to carry out the overall macroscopic circulation, and its macroscopic mixing intensity is weak, which will lead to a decrease in mass transfer effect. Uneven mixing of materials
The mass transfer effect of mixing will be worse in the reaction system with higher viscosity
[0014] When the gas-liquid dispersion rotor is placed in the middle of the tank, compared with the reactor with the gas-liquid dispersion rotor placed at the bottom of the tank, its critical speed is reduced, and the suction volume at the same speed is increased, but there will be gas inclusions in the upper part of the tank. The phenomenon that the rate is greater than the gas holdup rate in the lower part of the tank, and this phenomenon is extremely unfavorable for the growth of microorganisms

Method used

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  • High-dissolved oxygen bioreactor for high-density cultivation of genetically engineered bacteria and cultivation control method
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  • High-dissolved oxygen bioreactor for high-density cultivation of genetically engineered bacteria and cultivation control method

Examples

Experimental program
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Effect test

Embodiment

[0054] like Figure 1 to Figure 6 As shown, the high-dissolved oxygen bioreactor for the high-density cultivation of genetically engineered bacteria of the present embodiment comprises a tank body with feeding holes 4 in the upper part and a discharge port 15 in the lower part. The output shaft of the drive motor 6 is connected to the stirring paddle extending into the tank; the rotor drive motor 16 is arranged outside the bottom of the tank, and the output shaft of the rotor drive motor 16 is connected to the gas-liquid dispersion rotor 18 inside the tank to achieve gas-liquid dispersion. The rotor 18 has a ventilated inner chamber with an air inlet on the top and an air outlet on the bottom, and the air inlet is connected to a pressurized air source through a pipeline; The diversion groove of the water inlet and the water outlet is established; the gas-liquid dispersion stator 19 is located in the circumferential direction of the gas-liquid dispersion rotor 18, and the gas-l...

Embodiment example 1

[0096] The size of the reactor is: the volume of the tank is 5m 3 , the height-to-diameter ratio of the tank is 2.2:1, the diameter of the gas-liquid dispersion rotor is 215mm, the gas-liquid dispersion stator is designed with 16 diversion grooves, the outer diameter is 405mm, and the width of the four baffles is 140mm. There are two layers of stirring paddles with a diameter of 500 mm.

[0097] In this implementation case, under the condition of 2.8 tons of liquid, the critical air flow rate of the gas-liquid dispersion rotor at a speed of 2100r / min, the critical speed of the gas-liquid dispersion rotor at this air flow rate, and the maximum stirring speed were measured.

[0098] The measured critical ventilation volume is 134.4m 3 / h, according to 85% of the critical ventilation rate is 114.2m 3 / h air intake, the measured critical speed of the gas-liquid dispersion rotor is 1950r / min. The speed of the gas-liquid dispersion rotor is set to 120% of the critical speed, that ...

Embodiment example 2

[0103] The size of the reactor is: the volume of the tank is 5m 3 , the height-to-diameter ratio of the tank is 2.2:1, the diameter of the gas-liquid dispersion rotor is 215mm, the gas-liquid dispersion stator is designed with 16 diversion grooves, the outer diameter is 405mm, and the width of the four baffles is 140mm. There are two layers of stirring paddles with a diameter of 500 mm.

[0104] In this implementation case, under the condition of 2.8 tons of liquid, the critical air flow rate of the gas-liquid dispersion rotor at a speed of 2100r / min, the critical speed of the gas-liquid dispersion rotor at this air flow rate, and the maximum stirring speed were measured.

[0105] The measured critical ventilation volume is 134.4m 3 / h, according to 70% of the critical ventilation rate is 94m 3 / h air intake, the measured critical speed of the gas-liquid dispersion rotor is 1780r / min. The speed of the gas-liquid dispersion rotor is set to 120% of the critical speed, that is...

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Abstract

The invention relates to a high dissolved oxygen bioreactor for the high-density culture of genetically engineered bacteria and a culture control method. The reactor comprises a tank body, wherein the top of the tank body is provided with a stirring driving motor, and an output shaft of the stirring driving motor is used for driving a stirring paddle and is connected with the stirring paddle; the bottom of the tank body is provided with a rotor driving motor, an output shaft of the rotor driving motor is used for driving a gas-liquid dispersion rotor and is connected with the gas-liquid dispersion rotor, the gas-liquid dispersion rotor is provided with an inner ventilation cavity which is provided with an air inlet at the top and an air outlet at the lower part, and the air inlet is connected with an under-pressure air source through a pipeline; the inner side at the bottom of the tank body is also fixedly connected with a gas-liquid dispersion stator provided with a diversion trench; the gas-liquid dispersion stator is located at the circumference of the gas-liquid dispersion rotor and is coaxial with the gas-liquid dispersion rotor to form a rotating pair. The method comprises a gas-liquid dispersion rotor ventilation control process, a gas-liquid dispersion rotor rotating speed control process and a stirring paddle stirring rotating speed control process. The high dissolved oxygen bioreactor is high in dissolved oxygen level and suitable for the high-density culture of the genetically engineered bacteria.

Description

technical field [0001] The invention relates to a high-dissolved oxygen bioreactor for high-density cultivation of genetically engineered bacteria and a cultivation control method using the reactor, belonging to the technical field of biological fermentation engineering. Background technique [0002] With the development of genetic engineering technology, more and more biological products can be produced through genetic engineering process. Among them, Escherichia coli and Pichia pastoris have been widely used in the construction of genetically engineered bacteria to obtain exogenous gene products that are useful to people. [0003] The use of genetically engineered bacteria for fermentation aims to obtain large quantities of high-quality exogenous gene products, and at the same time reduce the contamination of the host cell's own components as much as possible, so its fermentation process is different from that of traditional microorganisms. [0004] The traditional batch ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/04C12M1/02C12Q3/00
CPCC12M27/02C12M29/06C12M29/14C12M41/00
Inventor 洪厚胜李军庆蔡子金
Owner NANJING TECH UNIV
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