Skip to main content

Biosensor-based dual-color droplet microfluidic platform for precise high-throughput screening of erythromycin hyperproducers

 



1. Introduction

The growing demand for natural products in pharmaceuticals, agriculture, and biotechnology has prompted advancements in microbial cell factory engineering. Among the innovative approaches, biosensor-based droplet microfluidic high-throughput screening has emerged as a powerful technique for detecting and selecting high-yield microbial strains. This method leverages genetically encoded biosensors to produce measurable outputs in response to specific metabolite concentrations, enabling rapid identification of desirable phenotypes from large mutant libraries. However, inherent biological variability among microbial cells poses challenges to the reliability and accuracy of this technique, necessitating refined strategies to enhance screening fidelity.

2. Limitations of Traditional Whole-Cell Biosensors in Droplet Microfluidics

Conventional single-color whole-cell biosensors, while effective under controlled conditions, often fail to maintain accuracy within microfluidic droplets. Environmental fluctuations significantly influence bacterial growth and gene expression, leading to heterogeneous cell populations within each droplet. This variability introduces inconsistencies in biosensor output signals, making it difficult to distinguish true positive signals from background noise. Moreover, the inability to measure or control cell density within individual droplets further exacerbates false-positive rates, creating a substantial burden for downstream validation processes.

3. Engineering Dual-Color Biosensors for Normalized Signal Output

To address the challenge of heterogeneity, this study introduced a novel dual-color biosensor design in Escherichia coli. By integrating a second reporter signal that reflects cell growth or viability, the system provides normalized outputs by comparing the product-indicative fluorescence to a constitutive reference. This internal control corrects for variances in cell density and gene expression, resulting in more accurate and robust detection of desired phenotypes. The dual-color format represents a significant step forward in biosensor design for high-throughput screening applications.

4. Integration with Droplet-Based Microfluidic Platforms

The enhanced dual-color biosensors were successfully implemented in a droplet-based microfluidic screening platform, enabling simultaneous analysis of thousands of individual microbial variants. This integration allowed for high-throughput, parallelized screening with improved signal consistency and droplet uniformity. In proof-of-concept experiments, the dual-color system exhibited a markedly higher enrichment ratio compared to its single-color counterpart, underscoring the effectiveness of this strategy in minimizing heterogeneity-induced noise and improving screening outcomes.

5. Application in Erythromycin-Producing Strain Improvement

To validate the practical benefits of the dual-color approach, the system was employed in screening both wild-type and mutagenized Saccharopolyspora erythraea strains for enhanced erythromycin production. Results showed a 24.2% increase in positive identification rates for the wild-type strain and an 11.9% increase for industrial S0-derived libraries using the dual-color method. Notably, strains exhibiting up to a 19.6% improvement in erythromycin yield were successfully isolated, demonstrating the method's potential for industrial strain development and optimization.

6. A Universal Strategy for Next-Generation Biosensor Applications

The dual-color whole-cell biosensor platform provides a universal framework for improving the accuracy and throughput of microbial screening campaigns. By accounting for biological variability within droplets, this system reduces false positives and minimizes the time and resources required for post-screening verification. Its compatibility with diverse natural product biosynthesis pathways makes it a versatile tool for synthetic biology, metabolic engineering, and bioprocess development. This work lays the foundation for the next generation of biosensor-driven high-throughput screening technologies.

🔬 Visit: biophotonicsresearch.com
🏅 Nominate Now: https://jif.li/GZxtt

#Microfluidics #Biosensors #SyntheticBiology #MetabolicEngineering #DualColorBiosensors #HighThroughputScreening #NaturalProductDiscovery #MicrobialCellFactories #StrainEngineering #ErythromycinProduction #DropletMicrofluidics #WholeCellBiosensors #CellHeterogeneity #GeneticallyEncodedSensors #FluorescenceScreening #BiotechnologyInnovation #BioengineeringTools #BioprocessOptimization #BiotechResearch #NextGenScreening


Comments

Post a Comment

Popular posts from this blog

Abrisa Technologies Acquires Agama Glass Technologies

SANTA PAULA, Calif. — Abrisa Technologies, a provider of custom glass optics and thin film coatings and a subsidiary of HEF Photonics, has acquired Agama Glass Technologies, a manufacturer of etched anti-glare glass and technical glass processing. The acquisition, Abrisa said, expands its manufacturing footprint and adds a vertically integrated solution for chemically etched anti-glare display glass. According to Abrisa, Clarksburg, West Virginia-based Agama operates North America’s only high-volume technical glass etching facility. Agama's flagship product, AgamaEtch, is used in high-performance display and optics applications. The company's 85,000 sq ft facility also offers precision glass fabrication, chemical strengthening, and silk-screen printing, serving markets such as avionics, defense, medical, industrial, and touchscreen displays. Combined with Abrisa Technologies’ and HEF Photonics’ thin-film coating and surface engineering capabilities, Agama's offerings wi...
Post a Comment
Read more

How Biophotonics Is Harnessing Light for Health And Science

Fifty or so years ago French physicist Pierre Aigrain coined the term photonics as a research field whose goal was to use light to perform functions that traditionally fell within the typical domain of electronics, such as telecommunications, and information processing. Or maybe it was John Campbell who, in a letter sent to Gotthard Gunther in 1954, wrote, “Incidentally, I’ve decided to invent a new science — photonics. It bears the same relationship to Optics that electronics does to electrical engineering. Photonics, like electronics, will deal with the individual units; optics and EE deal with the group phenomena! And note that you can do things with electronics that are impossible in electrical engineering!” Naming rights aside, the field of photonics began in earnest between 1958 and 1960 with the invention of the maser and the laser. The laser diode followed during the 1970s, optical fibers and the erbium-doped fiber amplifier after that, and, pretty soon, the telecommunications...
Post a Comment
Read more

Laser Method Enables Fast & Precise Blood Vessels in Hydrogel

Researchers from Vienna University of Technology (TU Wien) and Keio University have found a way to create artificial blood vessels in miniature organ models in a quick and reproducible manner. The method utilizes ultrashort laser pulses in the femtosecond range to write highly 3D structures into a hydrogel. In biomedical research, organs-on-a-chip are becoming increasingly important: By cultivating tissue structures in precisely controlled microfluidic chips, it is possible to conduct research much more accurately than in experiments involving living humans or animals. However, there has been a major obstacle: such mini-organs are incomplete without blood vessels. To facilitate systematic studies and ensure meaningful comparisons with living organisms, a network of perfusable blood vessels and capillaries must be created — in a way that is precisely controllable and reproducible. “We can create channels spaced only a hundred micrometers apart. That’s essential when you would like to...
Post a Comment
Read more