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An expert control system for expressing foreign proteins in recombinant Pichia pastoris

An exogenous protein, Pichia pastoris technology, applied in animal/human protein, fermentation, microorganism-based methods, etc., can solve problems such as difficulty in achieving satisfactory results

Active Publication Date: 2017-04-12
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the protein expression system of Pichia pastoris, several important control variables, such as temperature, DO, sorbitol flow rate (blending induction), etc., have a very complicated relationship with the physiological state of the cell. Difficult to achieve satisfactory results

Method used

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  • An expert control system for expressing foreign proteins in recombinant Pichia pastoris
  • An expert control system for expressing foreign proteins in recombinant Pichia pastoris
  • An expert control system for expressing foreign proteins in recombinant Pichia pastoris

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Application of Expert Control System in the Process of Recombinant Pichia Pastoris Fermentative Production of Porcine Interferon Alpha

[0054] The expert control system for the process of fermenting and producing porcine interferon alpha by recombinant Pichia pastoris, the multi-variable on-line metering and control device for the fermentation process is shown in the figure figure 1 shown.

[0055] Here, the state variable methanol concentration is individually controlled by methanol online measuring electrodes (concentration level fixed). The other 3 control variables directly related to fermentation performance, fermentation temperature (I / O switch controls cooling water bath temperature), sorbitol flow acceleration (D / A controls peristaltic pump) and stirring speed (two-way communication, D / A controls stirring motor ), through online measurement of oxygen uptake rate OUR / carbon dioxide release rate CER (RS232 data acquisition) and methanol consumption rat...

Embodiment 2

[0068] Example 2 Application of expert control system in the process of recombinant Pichia pastoris fermentation and production of Cap protein

[0069] The expert control system for the fermentation and production of Cap protein by recombinant Pichia pastoris, the multi-variable on-line metering and control device for the fermentation process is shown in Fig. figure 1 shown.

[0070] where r (C) NADH The division of , DO and R is shown in Table 2. δ in formulas (5), (6), (7) 1 ,δ 2 ,δ 3 1, 0.15, 20 respectively, sorbitol initial flow acceleration rate r SOR(0), initial temperature T(0) and initial stirring speed r AG (0) were 1 g / L / h, 30°C and 400 rpm, respectively. The control period is 15min. Other specific implementations are the same as in Example 1. After activation, the Pichia pastoris GS115 strain was inoculated into a 10L fermenter according to the inoculation amount of 20%, and then induced after culturing for 30 hours at 30°C, pH 6.0, and ventilation volume...

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Abstract

The invention discloses an expert control system for a process of expressing a foreign protein by recombinant pichia pastoris and belongs to the biotechnical field. When pichia pastoris expresses the foreign protein, several important control variables such as temperature, DO, sorbitol feeding speed (blend induction) and the like have a very complicated relationship with physiological statuses of cells. Aiming at the complicated process that the control variables are mutually interfered, the quantitative relation between the control variables and the controlled variables is not clear and the performance indexes are hard to be independently controlled, the invention provides an expert control strategy. The expert control system is an intelligent computer control system which internally contains expert knowledge and experience. The expert control system is used for solving related problems in a compound optimized control process by constructing, implementing and adjusting expert commands and rules. The control system controls the fermenting temperature, the sorbitol feeding speed and dissolved oxygen online in the optimum level simultaneously by an energy-saving way, so that the activity and concentration of a target protein are improved and the operating cost and energy consumption are lowered.

Description

technical field [0001] The invention relates to an expert control system for the process of expressing exogenous protein by recombinant Pichia pastoris, belonging to the field of biotechnology. Background technique [0002] Domestic and foreign research reports on the optimization and control of recombinant Pichia pastoris in the induction stage mainly focus on univariate (factor) control to enhance the synthesis and accumulation of target proteins, but there is basically no overall optimization of the fermentation process (improvement of fermentation performance + reduction of operating energy consumption). consider. However, in the protein expression system of Pichia pastoris, several important control variables, such as temperature, DO, sorbitol flow rate (blending induction), etc., have a very complicated relationship with the physiological state of the cell. Difficult to achieve satisfactory results. Only on the basis of analyzing the individual effects of each operat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/38C12M1/02C12P21/00C12R1/865
CPCC07K14/56C12M27/02C12M41/00C12M41/12C12M41/22C12M41/32C12M41/34C12M41/48
Inventor 高敏杰詹晓北史仲平丁健杨帅
Owner JIANGNAN UNIV
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