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Escherichia coli is widely used as a workhorse for heterologous protein expression. However, insoluble protein expression is frequently encountered. The mechanisms underlying protein solubility remain incompletely understood, limiting the effectiveness of current protein solubility engineering strategies. Here, SeqSo...
Efficient electron supply and cofactor utilization remain key bottlenecks in cytochrome P450-catalyzed oxidations. Here, we report an electro-assisted system that overcomes these limitations, using the P450 Ema1-catalyzed oxidation of avermectin B1a as a model reaction. Under a constant current of 1 mA, > 96% conversio...
l-Ornithine is a versatile amino acid with broad applications in the pharmaceutical, nutraceutical, and food industries. However, its efficient microbial production is constrained by a critical trade-off: excessive blockage of the downstream l-arginine pathway severely impairs cell growth, while intracellular l-ornit...
Microbial production of l-arginine is often constrained by tight metabolic regulation and insufficient precursor supply. Here a plasmid-free, non-auxotrophic Escherichia coli strain for high-level production of l-arginine was rationally engineered, based on our previous constructed strain G0 with 12.4 g/L l-arginine pr...
The integration of artificial intelligence with protein engineering has created unprecedented opportunities for enzyme discovery and the enhancement of overall catalytic performance. In this study, a novel nitrilase, BhNIT, was identified from Bradyrhizobiumhuanghuaihaiense through a computationally assisted screenin...
d-Pantothenic acid (D-PA) is an essential precursor of coenzyme A and the acyl carrier protein. Its biosynthesis requires abundant precursors and precise regulation of the multifunctional enzyme, ketopantoate reductase. Here, we engineered Corynebacterium glutamicum for enhanced D-PA production through metabolic engi...
This study demonstrated that the GA4/GA7 ratio can be modulated through both molecular and fermentation strategies, and engineering a high-yielding GA4+7 producer using this Δp450-3 mutant as a platform.
The plasmid-free system constructed in this study effectively avoids the plasmid-induced metabolic burden and genetic instability and demonstrates the prominent advantages and great application potential of plasmid-free modular engineering for the efficient biosynthesis of OSH and other high-value amino acids.
This study establishes a robust chassis for sustainable DPA production and provides a generalizable framework for engineering other valuable biochemicals.
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