The protein journal最新文献

Protein and peptide drugs have become essential in treating numerous diseases due to their high specificity and therapeutic potential. However, their clinical application faces several challenges, including chemical instability, physical instability, short in vivo half-life, low oral bioavailability, and immunogenicity. These issues reduce drug efficacy and limit patient compliance. This review critically examines these limitations and presents current strategies to address them. PEGylation, the covalent or noncovalent attachment of polyethylene glycol (PEG) molecules to proteins, is highlighted for its ability to increase stability, reduce renal clearance, lower immunogenicity, and extend half-life. The review distinguishes between random and site-specific PEGylation, highlighting site-specific methods that preserve protein activity while optimizing pharmacokinetics. Additionally, the encapsulation of proteins in polymeric and lipid-based delivery systems is discussed to protect drugs from enzymatic degradation, control their release, and enhance absorption. Biobetter approaches, including site-directed mutagenesis, are also presented to improve protein stability and reduce immunogenicity. Market data and approved drug examples illustrate the impact of these strategies. Overall, the article provides a comprehensive overview of innovative pharmaceutical and biotechnological methods that enhance the clinical performance and market viability of protein therapeutics.

The utilization of Bacillus spp. as eco-compatible bioresources represented a promising strategy in antifungal biotechnology. In this study, Bacillus subtilis Bs-06, isolated from the rhizosphere of healthy tomato plants, exhibited strong antagonistic activity through the secretion of diverse bioactive metabolites. Gas chromatography-mass spectrometry (GC-MS) analysis of its culture extract identified ten major compounds, including Phenol-3,5-bis(1,1-dimethylethyl), palmitic acid, oleic acid, and octadecanoic acid, which had been widely reported for antimicrobial efficacy. To investigate their Molecular Docking targeted the key virulence proteins of Fusarium oxysporum f.sp. lycopersici Cutinase (5AJH), Avr1/SIX4 (7T6A), and Avr3/SIX1 (7T69). Docking simulations revealed strong binding affinities, with Phenol-3,5-bis(1,1-dimethylethyl) showing the highest interaction energy (- 11.53 kcal/mol) against Avr3. The ligand-protein complexes were stabilized by hydrogen bonding and hydrophobic interactions, suggesting potential inhibition of pathogen virulence functions. Normal mode analysis further demonstrated differential flexibility among protein-ligand complexes, where Avr1 exhibited the highest conformational mobility, while Cutinase displayed the greatest structural rigidity. These findings established Bs-06 as a valuable source of antifungal metabolites capable of targeting virulence determinants at the molecular level. The integration of GC-MS profiling, molecular docking, and dynamic simulations underscored the potential of Bacillus derived metabolites as sustainable biocontrol agents for managing Fusarium wilt in tomato.

Sea cucumbers Eupentacta fraudatrix can completely regenerate their organs within one month after evisceration. The regeneration process involves complex structural changes including restructuring of the intracellular matrix. The connective tissue of echinoderms consists of bundles of collagen fibrils and proteoglycans. It is assumed that animals capable of regeneration must possess a diverse set of proteases and glycosylases that modify the connective tissue. Glycosidases catalyze the hydrolysis of glycosidic bonds in carbohydrate molecules, and these enzymes have not yet been studied in the sea cucumber E. fraudatrix. Here four glycosidase-enriched fractions having optimal pH values at 7.0, 7.5 (two fractions), and 6.0 were revealed. Thе glycosidase of Peak 1 (pH 7.0) was moderately activated by Mn²⁺, Ca²⁺, and Zn²⁺ but its best activators are Co²⁺ and Ni²⁺, while Mg²⁺ ions suppress glycosidase activity. Thе glycosidase of Peak 2 (pH 7.5) is activated at low concentrations of Ca²⁺ and Mg²⁺; Mn²⁺ and Co²⁺ increase its activity at higher concentrations, while Ni²⁺ and Zn²⁺ are inhibitors of this enzyme. Thе glycosidase of Peak 3 (pH 7.5) has maximal activity in the presence of Ca²⁺ and Mg²⁺ ions but its activity is suppressed by Mn²+, Ni²⁺, and Zn²⁺. Acid glycosidase (pH 6.0) is Ca²⁺ dependent enzyme, which activity suppresses by several metal ions to varying degrees: Zn²⁺ > Mg²⁺ > Ni²⁺ > Mn²⁺ > Co²⁺. These are the first glycosidases identified in the sea cucumber E. fraudatrix that can be utilized for future studies of their role in extracellular matrix remodeling.

Stonustoxin (SNTX) is a lethal protein found in stonefish venom, responsible for many of the symptoms associated with stonefish envenomation. To counter stonefish venom challenges, antivenom is a well-established and effective solution. In this study, we aimed to produce the recombinant alpha subunit protein of Stonustoxin from Synanceia horrida and prepare antibodies against it The SNTXα gene sequence was optimized for E. coli BL21 (DE3) expression and cloned into the pET17b vector. Following purification, the recombinant protein was subcutaneously injected into rabbits, and antibodies were extracted from rabbit´s serum using a G protein column As a result of codon optimization, the codon adaptation index for the SNTXα cassette increased to 0.94. SDS-PAGE analysis validated the expression of SNTXα, with a band observed at 73.5 kDa with a yield of 60 mg/l. ELISA results demonstrated rabbits antibody titers were detectable up to a 1:256,000 dilution. The isolated antibody from rabbit´s serum exhibited a concentration of 1.5 mg/ml, and its sensitivity allowed the detection of a minimum protein concentration of 9.7 ng. In the neutralization assay the purified antibody against SNTXα protected mice challenged with 2 LD50. In conclusion, our study successfully expressed the alpha subunit of Stonustoxin in a prokaryotic host, enabling the production of antibodies for potential use in developing stonefish antivenom.