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Related Experiment Video

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A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

Published on: July 31, 2012

Screening microorganisms with robust and stable protein expression and secretion capacity.

Li-Hua Liu¹, Yu Guo¹, Min Yang¹

¹Tidetron Bioworks Technology (Guangzhou) Co., Ltd., Guangzhou Qianxiang Bioworks Co., Ltd., Guangzhou, Guangdong, People's Republic of China.

Protein Science : a Publication of the Protein Society

|December 17, 2024

Summary

A new split GFP activated droplet sorting (SGADS) platform significantly enhances recombinant protein production. This method optimizes protein secretion and protease activity screening, boosting yields of valuable proteins.

Keywords:

droplet sorting protein secretion proteinase activity split GFP sensors

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Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

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Area of Science:

Biotechnology
Molecular Biology
Protein Engineering

Background:

Efficient recombinant protein production relies on robust protein secretion.
Optimizing secretion pathways and screening for high-yield strains are critical challenges.

Purpose of the Study:

To develop a novel platform, split GFP activated droplet sorting (SGADS), to enhance recombinant protein yields.
To create and validate split GFP sensors for assessing protease activity and secretion capacity.

Main Methods:

Developed the SGADS platform using solubilizing peptide P17 and secretory expression in Bacillus subtilis.
Engineered two split GFP sensors: P17-GFP1-9/GFP10-POI-GFP11 for protease activity and P17-GFP1-10/GFP11-POI for secretion capacity.
Applied SGADS to screen high-performance mutant strains across various host organisms.

Main Results:

Successfully screened high-performance mutant strains for producing collagen, amylase, and protein glutaminase.
Demonstrated significant increases in protein production yields using the SGADS platform.
Validated the platform's effectiveness in diverse host organisms like E. coli, B. subtilis, and P. pastoris.

Conclusions:

The SGADS platform is a powerful tool for boosting recombinant protein production yields.
The developed split GFP sensors effectively assess protease activity and secretion capacity.
SGADS shows broad applicability and potential for enhancing protein manufacturing across different hosts.