Sodium Pyruvate Addition in Pichia Fermentation for Collagen
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Solution Overview
Problem
Current fermentation processes for recombinant human collagen production using genetically engineered bacteria, such as Pichia yeast, face challenges of long fermentation periods and low production efficiency, which hinder industrialized large-scale production.
Innovation Solution
The process involves inoculating a sterilized fermentation medium with Pichia yeast and adding sodium pyruvate (0.01-10 g/L) during the methanol-induced expression stage, optimizing fermentation conditions like pH, temperature, and oxygen levels, and using a fed-batch mode to enhance the biosynthesis rate and shorten the fermentation time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional fermentation processes are used with genetically engineered bacteria, then the fermentation period is extended and production efficiency is reduced, but the process maintains stable operation
Solution Approach 1:
The patent changes the chemical composition parameters of the fermentation medium by adding sodium pyruvate (0.01-10 g/L) to optimize metabolic pathways. This parameter modification accelerates the fermentation process, reducing the fermentation period from traditional extended durations to 99 hours while simultaneously increasing collagen production efficiency and fermentation level to 0.051 g/L·h
Solution Approach 2:
Sodium pyruvate serves as a metabolic intermediary substance that bridges the gap between traditional fermentation limitations and desired high-efficiency production. It acts as a precursor that enhances the metabolic rate and growth of Pichia pastoris, enabling faster collagen synthesis without compromising process stability
2Duration of action of moving object
If Pichia pastoris C13 is used as a fungal strain, then the fermentation period is shortened to 99 hours, but the protein expression level and fermentation level are too low at 5 g/L and 0.051 g/L·h
Solution Approach 1:
The patent modifies the fermentation medium composition by introducing sodium pyruvate at optimized concentrations (0.01-10 g/L). This parameter change simultaneously addresses both the fermentation duration and protein expression level, achieving a balance where the fermentation period remains short (99 hours) while the collagen production reaches 5 g/L with an improved fermentation level of 0.051 g/L·h
3Quantity of substance
If fermentation period is extended to 136 hrs to increase protein expression level to 16 g/L, then the fermentation level remains low at 0.118 g/L·h
Solution Approach 1:
Sodium pyruvate acts as a metabolic intermediary that enhances the efficiency of protein synthesis per unit time. By providing readily metabolizable carbon and energy sources, it enables the system to achieve higher protein expression levels (16 g/L) without requiring extended fermentation periods (136 hrs), thereby improving the fermentation level to 0.118 g/L·h through more productive metabolism
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach significantly increases the expression and production levels of recombinant human collagen by more than 20%, reduces fermentation time, and lowers production costs, making it suitable for industrial-scale production.
Implementation Method 1
performing fermentation culturing for 14-18 hrs, and adding methanol to induce the expression
Implementation Method 2
the growth and metabolic rate of the Pichia yeast is regulated
Data Source
AI summary
The present invention provides a fermentation process for improving the production level of a recombinant human collagen. The process comprises inoculating a sterilized fermentation medium with a Pichia yeast solution, then performing fermentation culturing for 14-18 hrs, adding methanol to induce the expression, upon which sodium pyruvate is added to the fermentation medium, wherein the amount of the sodium pyruvate added is 0.01-10 g/L. In the fermentation process of the present invention, the sodium pyruvate is added in the methanol-induced expression stage, so that the biosynthesis rate of the recombinant human collagen is improved; and a continuous feed-batch mode is adopted, so that the biosynthesis rate of the recombinant human collagen is further improved, and the fermentation time is shortened. Meanwhile, the expression level of the recombinant human collagen is increased, the fermentation level is increased by more than 20%, and the production cost is reduced. The fermentation process is especially suitable for industrialized mass production of recombinant human collagen, and is of huge practical application value in industrial production.