Whole-Cell Biosensor and Producer Co-cultivation-Based Microfludic Platform for Screening <i>Saccharopolyspora erythraea</i> with Hyper Erythromycin Production

Hua, Erbing; Zhang, Yue; Yun, Kaiyue; Pan, Wenjia; Liu, Ye; Li, Shixin; Wang, Yan; Tu, Ran; Wang, Meng

doi:10.1021/acssynbio.2c00102

Cited by 12 publications

(15 citation statements)

References 40 publications

(67 reference statements)

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“…A two-plasmid biosensor strain Top10/MphR, where mphr and mpha genes were carried by pMphR and pMphA plasmids, respectively, was used for high-throughput screening of hyper erythromycin producers in our previous work . However, this biosensor had no response to the native producer S.…”

Section: Resultsmentioning

confidence: 99%

Modulating Sensitivity of an Erythromycin Biosensor for Precise High-Throughput Screening of Strains with Different Characteristics

Wang

Xue

et al. 2023

ACS Synth. Biol.

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Genetically encoded biosensors are powerful tools for product-driven high-throughput screening in synthetic biology and metabolic engineering. However, most biosensors can only properly function in a limited concentration cutoff, and the incompatible performance characteristics of biosensors will lead to false positives or failure in screening. The transcription factor (TF)-based biosensors are usually organized in modular architecture and function in a regulator-depended manner, whose performance properties can be fine-tuned by modifying the expression level of the TF. In this study, we modulated the performance characteristics, including sensitivity and operating range, of an MphR-based erythromycin biosensor by fine-adjusting regulator expression levels via ribosome-binding site (RBS) engineering and obtained a panel of biosensors with varied sensitivities by iterative fluorescence-assisted cell sorting (FACS) in Escherichia coli to accommodate different screening purposes. To exemplify their application potential, two engineered biosensors with 10-fold different sensitivities were employed in the precise high-throughput screening by microfluidic-based fluorescence-activated droplet sorting (FADS) of Saccharopolyspora erythraea mutant libraries with different starting erythromycin productions, and mutants representing as high as 6.8 folds and over 100% of production improvements were obtained starting from the wild-type strain and the high-producing industrial strain, respectively. This work demonstrated a simple strategy to engineer biosensor performance properties, which was significant to stepwise strain engineering and production improvement.

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Section: Resultsmentioning

confidence: 99%

Modulating Sensitivity of an Erythromycin Biosensor for Precise High-Throughput Screening of Strains with Different Characteristics

Wang

Xue

et al. 2023

ACS Synth. Biol.

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“…The diversity of porous materials stems from a wide range of preparation methods [ 50 , 51 , 52 , 53 , 54 ]. Because of their controllable pore size and porosity, porous materials offer a wide range of applications for microfluidic systems [ 9 , 55 , 56 , 57 , 58 , 59 ].…”

Section: Different Materials and Preparation Methods For Microfluidic...mentioning

confidence: 99%

Porous Structural Microfluidic Device for Biomedical Diagnosis: A Review

et al. 2023

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Microfluidics has recently received more and more attention in applications such as biomedical, chemical and medicine. With the development of microelectronics technology as well as material science in recent years, microfluidic devices have made great progress. Porous structures as a discontinuous medium in which the special flow phenomena of fluids lead to their potential and special applications in microfluidics offer a unique way to develop completely new microfluidic chips. In this article, we firstly introduce the fabrication methods for porous structures of different materials. Then, the physical effects of microfluid flow in porous media and their related physical models are discussed. Finally, the state-of-the-art porous microfluidic chips and their applications in biomedicine are summarized, and we present the current problems and future directions in this field.

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“…The highest product titer reached 1.836 g/L, representing a 660% improvement [ 7 ]. For biosensor engineering, the microfluidic platform and biosensor engineering were combined to screen erythromycin-producing strains of Saccharopolyspora erythraea , and a 50% improvement of the erythromycin yield was obtained [ 39 ]. In addition, promoter libraries can assist the optimization of genetic circuit.…”

Section: Application Of Filamentous Bacteria As Microbial Cell Factoriesmentioning

confidence: 99%

Microbial cell factories based on filamentous bacteria, yeasts, and fungi

Ding

2023

Microb Cell Fact

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Background Advanced DNA synthesis, biosensor assembly, and genetic circuit development in synthetic biology and metabolic engineering have reinforced the application of filamentous bacteria, yeasts, and fungi as promising chassis cells for chemical production, but their industrial application remains a major challenge that needs to be solved. Results As important chassis strains, filamentous microorganisms can synthesize important enzymes, chemicals, and niche pharmaceutical products through microbial fermentation. With the aid of metabolic engineering and synthetic biology, filamentous bacteria, yeasts, and fungi can be developed into efficient microbial cell factories through genome engineering, pathway engineering, tolerance engineering, and microbial engineering. Mutant screening and metabolic engineering can be used in filamentous bacteria, filamentous yeasts (Candida glabrata, Candida utilis), and filamentous fungi (Aspergillus sp., Rhizopus sp.) to greatly increase their capacity for chemical production. This review highlights the potential of using biotechnology to further develop filamentous bacteria, yeasts, and fungi as alternative chassis strains. Conclusions In this review, we recapitulate the recent progress in the application of filamentous bacteria, yeasts, and fungi as microbial cell factories. Furthermore, emphasis on metabolic engineering strategies involved in cellular tolerance, metabolic engineering, and screening are discussed. Finally, we offer an outlook on advanced techniques for the engineering of filamentous bacteria, yeasts, and fungi.

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Whole-Cell Biosensor and Producer Co-cultivation-Based Microfludic Platform for Screening Saccharopolyspora erythraea with Hyper Erythromycin Production

Cited by 12 publications

References 40 publications

Modulating Sensitivity of an Erythromycin Biosensor for Precise High-Throughput Screening of Strains with Different Characteristics

Modulating Sensitivity of an Erythromycin Biosensor for Precise High-Throughput Screening of Strains with Different Characteristics

Porous Structural Microfluidic Device for Biomedical Diagnosis: A Review

Microbial cell factories based on filamentous bacteria, yeasts, and fungi

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