Enhancing single-cell hyaluronic acid biosynthesis by microbial morphology engineering.

Area of Science:

• Synthetic Biology
• Microbial Engineering
• Bioprocess Technology

Background:

• Microbial morphology engineering offers a novel strategy to enhance product titers in bio-manufacturing.
• Hyaluronic acid (HA), a valuable glycosaminoglycan, is produced by HA synthase (HAS).
• Modulating cell shape is explored to improve single-cell HA-producing capacity.

Purpose of the Study:

• To engineer cell morphology in C. glutamicum for enhanced hyaluronic acid (HA) production.
• To investigate the roles of DivIVA and FtsZ proteins in cell shape and HA production.
• To improve the single-cell HA-producing capacity of microbial cell factories.

Main Methods:

• Dual regulation (down/up) of DivIVA and FtsZ using promoter substitution or plasmid overexpression in C. glutamicum.
• Observation of varied cell morphologies including small-ellipsoid-like, bulb-like, long-rod, and dumbbell-like shapes.
• Analysis of HA production, cell growth, and HA synthase (HAS) membrane levels via flow cytometry.

Main Results:

• Reduced expression of DivIVA and FtsZ inhibited cell growth.
• FtsZ overexpression resulted in a long, thick rod shape with a 5.2-fold increase in surface area.
• Single-cell HA production capacity was improved by 13.5-fold, with a 2.1-fold increase in membrane-bound HAS.

Conclusions:

• FtsZ-mediated microbial morphology engineering significantly enhances hyaluronic acid production.
• Engineering cell shape is a valuable strategy for improving the efficiency of microbial cell factories.
• This approach holds potential for high-titer synthesis of various metabolic products.