A High-Throughput Cell-Free Enzyme Screening System Using Redox-Responsive Hydrogel Beads as Artificial Compartments

ACS Synthetic Biology, Год журнала: 2025, Номер unknown

Опубликована: Фев. 13, 2025

We have developed a rapid, simple, and high-throughput screening system for recombinant enzymes using disulfide-bonded hydrogel beads (HBs) produced via microfluidic method. These redox-responsive HBs were compatible with the biosynthesis of enzyme mutants cell-free protein synthesis, fluorescent staining through an enzymatic reaction, genetic information recovery after fluorescence-activated droplet sorting (FADS). The expression microbial transglutaminase zymogen (MTGz) synthesis cross-linking-reactivity-based product validated. Next-generation sequencing (NGS) analysis genes recovered from highly identified novel mutation sites (N25 N27) in propeptide domain. introduction these mutations allowed design engineered active MTGz, demonstrating potential as artificial compartments FADS-based selection that catalyze peptide cross-linking reactions.

Язык: Английский

---

Living Coral Displays, Research Laboratories, and Biobanks as Important Reservoirs of Chemodiversity with Potential for Biodiscovery

Marine Drugs, Год журнала: 2025, Номер 23(2), С. 89 - 89

Опубликована: Фев. 19, 2025

Over the last decades, bioprospecting of tropical corals has revealed numerous bioactive compounds with potential for biotechnological applications. However, this search involves sampling in natural reefs, and is currently hampered by multiple ethical technological constraints. Living coral displays, research laboratories, biobanks offer an opportunity to continue unravel chemodiversity, acting as "Noah's Arks" that may support molecules interest. This issue even more relevant if one considers reefs face unprecedent threats irreversible losses impair biodiscovery new products, processes, services. displays provide controlled environments studying producing both known metabolites under varied conditions, they are not prone common bottlenecks associated such loss source replicability. Research laboratories focus on a particular species or compound using were cultured ex situ, although differ from wild conspecifics metabolite production quantitative qualitative terms. Biobanks collect preserve specimens, tissues, cells, and/or information (e.g., genes, microorganisms), which offers plethora data study compounds' mode action without having cope issues related access, standardization, regulatory compliance. Bioprospecting these settings faces several challenges opportunities. On hand, it difficult ensure complexity highly biodiverse ecosystems shape chemodiversity corals. other possible maximize biomass fine tune synthesis interest environments. Collaborative efforts needed overcome barriers foster opportunities fully harness before in-depth knowledge pool irreversibly lost due reefs' degradation.

Язык: Английский

---

The biosynthesis of trillin 6’-O-glucoside: a low-abundance yet pharmacologically active polyphyllin from Paris polyphylla

Synthetic and Systems Biotechnology, Год журнала: 2025, Номер 10(2), С. 610 - 619

Опубликована: Март 1, 2025

Язык: Английский

---

Accelerating enzyme discovery and engineering with high-throughput screening

Natural Product Reports, Год журнала: 2024, Номер unknown

Опубликована: Янв. 1, 2024

Recent progress in the DBTL cycle, including machine learning, facilitated enzyme mining for biocatalysis. Automation and standardization of library construction, coupled to high-throughput screening, further accelerates discovery process.

Язык: Английский

---