Nanoscale surface engineered living cells with extended substrate spectrum.

Area of Science:

• Biotechnology
• Synthetic Biology
• Microbial Engineering

Background:

• Living microorganisms have limited substrate utilization capabilities.
• Surface engineering offers a route to enhance microbial functions.
• Layer-by-Layer (LbL) technology provides a versatile platform for nanoscale assembly on cell surfaces.

Purpose of the Study:

• To engineer the surface of the yeast Arxula adeninivorans LS3 (Arxula) using LbL technology.
• To extend the substrate spectrum of Arxula by introducing new enzymatic functions.
• To evaluate the stability of the engineered cell surface and enzyme conjugates.

Main Methods:

• Utilized Layer-by-Layer (LbL) assembly to encapsulate Arxula cells with polyelectrolyte and enzyme layers.
• Incorporated lactate oxidase enzyme into the outer layer of the capsule.
• Assessed the retention of biological activity post-encapsulation.
• Investigated the stability of the polymeric capsule and enzyme conjugates.

Main Results:

• Successful encapsulation of Arxula cells using LbL technology while preserving biological activity.
• Engineered Arxula demonstrated the ability to convert lactate to pyruvate via immobilized lactate oxidase.
• Extended the substrate utilization range of Arxula.
• Demonstrated stability of the engineered cell surface and enzyme linkages.

Conclusions:

• LbL technology is effective for surface engineering of living yeast cells.
• Surface-functionalized Arxula exhibits enhanced metabolic capabilities by extending its substrate spectrum.
• The engineered system offers a stable platform for developing novel biocatalytic microorganisms.