Bioprospecting microbial genomes to expand the biocatalytic toolbox of rubber oxygenases.

Surveying a Pseudomonas aeruginosa-derived oxidoreductase activity

Findings suggest that the studied FPMO may play a role in antibiotic resistance in P. aeruginosa by oxidatively inactivating ampicillin by oxidatively inactivating ampicillin.

Expression and biochemical characterization of a patatin-like medium-chain esterase Rv1063c from Mycobacterium tuberculosis

Mycobacterium tuberculosis is extremely dependent upon lipid-hydrolyzing enzymes to utilize the resources of host lipids and survive within the cell. Rv1063c is a conserved hypothetical protein predicted to share sequence homology with the patatin-like family. In the present study, the rv1063c gene was cloned and heterologously expressed in Escherichia coli BL21 (DE3). Functional enzyme was obtained by solubilizing inclusion bodies in 8 mol·L −1 urea, refolding via gradient dialysis, and purifying through Ni-affinity chromatography. Bioinformatic homology analysis indicates Rv1063c is homologous to members of the patatin-like family containing a putative Ser52-Asp166 catalytic dyad, though this residue pair has not been experimentally validated in the present study. The recombinant Rv1063c was mainly expressed as inclusion bodies without significant soluble cytoplasmic expression. Biochemical assays demonstrated the recombinant protein exclusively hydrolyzes medium-chain pnitrophenyl esters (C8–12), with maximum hydrolytic activity toward p-NP-C8 (caprylate), followed by C10 and C12, and exhibited the highest activity at 40 °C and pH 7.0. The enzyme exhibited moderate catalytic efficiency but poor thermal stability. All catalytic data in this paper were obtained from artificial p-nitrophenyl ester substrates, which cannot fully reflect natural lipid substrate preference of mycobacteria in vivo. This work biochemically characterizes Rv1063c as a medium-chain-specific esterase encoded by a protein homologous to the patatin superfamily. Its physiological and pathogenic roles remain undetermined, and further multi-level experiments are required to clarify its biological functions.

Genome mining of Trp-anthranilic acid cyclic dipeptides in Aspergillus driven by analysis of NRPS adenylation domains

A novel family of fungal protein biosurfactants: Discovery and sustainable production.

A new class of fungal biosurfactant proteins is introduced and a simplified downstream process based on methanol/chloroform extraction is developed, reducing costs while preserving functionality.

Screening of Actinomycetes from Rhizosphere Soils for L-glutaminase Production

The integrated experimental and in silico approach highlights soil-derived actinomycetes as versatile and sustainable bioresources with significant pharmaceutical and biotechnological potential, emphasizing their role in combating antimicrobial resistance and enabling the rational development of novel therapeutic and industrial products.

Mining, Cloning, and Bioinformatic Analysis of Amidase Genes from Three Brewing Microorganisms

Ethyl carbamate (EC), a Group 2A carcinogen, is unavoidably formed during fermentation and poses a persistent safety concern in alcoholic beverages and fermented foods. Although enzymatic degradation offers a mild and substrate-specific strategy, EC hydrolases suitable for brewing environments remain scarce, and heterologous expression of known candidates often results in inactive inclusion bodies. In this study, we mined amidase genes from three brewing-associated microorganisms—Bacillus velezensis, Clavispora lusitaniae, and Saccharomyces cerevisiae—based on the conserved GGSSGG motif of the AS-family amidases. Three full-length genes, amiE (1458 bp), amdA (1632 bp), and amd08 (1665 bp), were successfully cloned into pET-28a vectors. Prokaryotic expression revealed that AmiE (52.63 kDa) and AmdA (60.76 kDa) were predominantly deposited as insoluble inclusion bodies, while Amd08 (61.39 kDa) was not detected due to apparent gene silencing. To circumvent these expression barriers, we performed comprehensive bioinformatic analyses. All three proteins were predicted as stable, hydrophilic molecules with theoretical pI values of 5.12–5.47 and negative GRAVY indices. Secondary structures were dominated by α-helices and random coils, and homology modeling confirmed the presence of intact GGSSGG motifs and Lys-Ser-Ser catalytic triads in each protein. Multidimensional model validation further supported their stereochemical reliability. Collectively, this study enriches the genetic reservoir of brewing-origin EC hydrolases and provides essential molecular and structural foundations for future solubility engineering and enzyme preparation development.