Expanding the recombinant protein quality in Lactococcus lactis

Area of Science:

• Microbiology
• Biotechnology
• Protein Engineering

Background:

• Escherichia coli faces challenges in producing soluble recombinant proteins due to conformational stress.
• Generally Recognized As Safe (GRAS) bacterial hosts, like Lactococcus lactis, present an alternative with potentially higher protein solubility.
• Further analysis of recombinant protein quality in L. lactis is needed to assess its full potential.

Purpose of the Study:

• To evaluate the conformational quality and solubility of a recombinant protein in Lactococcus lactis.
• To identify optimal production conditions for improved recombinant protein yield and quality.

Main Methods:

• Production of an aggregation-prone GFP variant (VP1GFP) in L. lactis.
• Analysis of protein conformational quality using specific fluorescence emission.
• Investigation of the impact of production time, culture conditions, and growth temperature on protein characteristics.

Main Results:

• Production parameters significantly affect protein yield, solubility, and conformational quality.
• Fermentative metabolism and anaerobic growth conditions favor protein solubility and quality.
• Growth temperature plays a crucial role in regulating protein solubility and conformational quality.

Conclusions:

• Metabolic regime and cultivation temperature are key factors influencing recombinant protein solubility and quality in L. lactis.
• Anaerobic growth is optimal for recombinant protein production in this system.
• L. lactis demonstrates versatility for improving recombinant protein functionality, yield, and quality, highlighting its potential as a cell factory.