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Deacetylases can directly hydrolyze amide bonds for the environmentally friendly synthesis of the herbicide intermediate O-(3-chloro-2-propenyl)hydroxylamine. However, the rigid halogenated chain of N-[(E)-3-chloroprop-2-enoxy]acetamide (NECA) may hinder productive substrate positioning and limit catalytic efficiency....
The stability-activity trade-off remains a major bottleneck in engineering robust biocatalysts for industrial applications. Here, we developed an integrated computational strategy to simultaneously enhance the catalytic activity and thermostability of 7α-hydroxysteroid dehydrogenase (7α-HSDH) by engineering highly flex...
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...
This paper systematically reviewed the latest research progress on the closed-loop intelligent fermentation control systems, spanning from underlying data perception to high-level decision-making and the transformation of fermentation control from traditional "macroscopic feeding regulation" to "microscopic metabolic p...
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.
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