Archives of Microbiology最新文献

Mycoplasma genitalium, Ureaplasma urealyticum and Mycoplasma hominis are bacterial pathogens found in the genitourinary tract, implicated in a range of infections. In women, these infections including pelvic inflammatory disease, vaginitis, infertility, and cervical cancer, while in men, they can cause non-gonococcal urethritis, prostate cancer, among other conditions. These infections are a global health concern, with China identified as a country with a high prevalence. This review provides a comprehensive overview of the epidemiology, causative factors, and diagnostic methods for these three Mycoplasma species with in China. The rise of multi-drug resistance, driven by antibiotics overuse, poses a significant challenge to treatment, complicating patient management. These Mycoplasma species employ unique adhesion mechanisms that trigger a cascade of signal transduction, culminating to inflammatory responses, tissue damage, and the release of toxic metabolites. Here, we delineate the mechanisms of underlying Mycoplasma resistance and propose key therapeutic strategies for these three mycoplasmas in China. This includes a summary of effective antibiotic treatment strategies, and potential combinations of therapeutic to improve cure rates, and a discussion of potential therapeutic approaches using traditional Chinese medicine.

The NADH/NAD⁺ balance plays a critical role in regulating cellular and metabolic pathways. In Saccharomyces cerevisiae, glycerol-3-phosphate dehydrogenase (ScGPD) enzymes are essential for NADH homeostasis, glycerol biosynthesis, and osmotic stress adaptation. This study investigates the replacement of ScGPD isoforms with the water-forming NADH oxidase from Lactococcus lactis (LlnoxE) and its effects on 10% glucose fermentation dynamics in minimal medium under microaerobic conditions. We engineered S. cerevisiae strains by individually or sequentially deleting or substituting ScGPD isoforms with LlnoxE, generating strains with varying NADH oxidation levels, fermentation rates, and byproduct formation. The engineered strains exhibited three distinct fermentation profiles: faster strains (∆GPD2 and ∆GPD1,2), five medium-speed strains (native, ∆GPD1, LlnoxE/∆GPD1, LlnoxE/∆GPD2, and LlnoxE with GPD), and three slower strains (LlnoxE/∆GPD1,2, LlnoxE/∆GPD1-∆GPD2, and LlnoxE/∆GPD2-∆GPD1). Increased NADH oxidation correlated strongly with higher acetic acid production, which inhibited cell growth and reduced fermentation speed, especially when glycerol biosynthesis was abolished. For instance, LlnoxE/ΔGPD1 reduced glycerol production by 88% and increased ethanol yield by 6.2%, despite a 9% increase in acetic acid production. This study underscores the importance of NADH oxidation in optimizing fermentation efficiency and metabolic balance in S. cerevisiae strains lacking GPD during glucose fermentation.

Prions are proteinaceous infectious particles implicated in fatal neurodegenerative disorders known as prion diseases. Herein, we provide an overview of prion biology, emphasizing the structural, functional, and evolutionary aspects of prions, along with their potential applications in protein engineering. Understanding the structure-function relationships of both healthy and disease-associated prion proteins enables a deeper understanding of the mechanisms of prion-induced neurotoxicity. Furthermore, we describe how insights into prion evolution have begun to shed light on their ancient origins and evolutionary resilience, offering deeper insights into the potential roles of prions in primordial chemical processes.

Azurin, a secondary metabolite from Pseudomonas aeruginosa, has attracted much attention owing to its valuable therapeutic and biological applications. This work aimed to study and chartly maximize the azurin production process using different doses of gamma irradiation (5–400 Gy) in P. aeruginosa isolates. Seventy-six P. aeruginosa isolates were sourced from 135 environmental samples and 35 clinical bacterial isolates with the following descending order: 35 isolates (46%) from clinical samples, 26 isolates (34%) from water samples, and 15 isolates (20%) from soil samples. The disc diffusion technique was used for antimicrobial susceptibility testing, revealing that the multidrug-resistant (MDR) rate among all collected isolates according to the criteria determined by Clinical and Laboratory Standards Institute (CLSI) was 54 (71%). The genomic experimental results revealed that only 37 MDR isolates tested positive for the azurin gene, as detected by the PCR product at 446 bp. These findings were further supported by FTIR analysis, which revealed peaks around 1636.96 cm^− 1, indicating a prominent α-helix secondary structure of azurin in these isolates. Related to their pathogenicity and antibiotic resistance, isolates from clinical origin exhibited the higher azurin gene expression level. Besides, this study confirmed the potency of gamma radiation exposure at 50 and 100 Gy significantly increased the azurin expression levels in three tested clinical isolates (P ≤ 0.05), with a maximum fold expression level of 63.55 compared to the non-irradiated samples. In conclusion, low doses of gamma irradiation effectively enhanced expression level of a secondary metabolite azurin, providing a considerable benefit for subsequent purification processes in both biological and medical applications.

RfaH is a crucial protein involved in anti-termination of transcription, which is necessary for spreading virulence in certain types of bacteria, such as Klebsiella pneumoniae and Escherichia coli. RfaH works by interacting directly with RNA polymerase and ribosomes, which activates the production of certain components needed for the bacteria's survival. Targeting RfaH offers a novel approach to hindering bacterial transcription and virulence. In this study, we performed computational screening of the IMPPAT 2.0 database consisting of 17,967 natural compounds, which were filtered based on Lipinski’s RO5 filter, selecting only those that had druglike properties. We performed virtual screening on the remaining 11,708 druglike phytochemicals and selected those having strong binding affinity and specificity, leading to the identification of top hits. These hits were further evaluated based on their pharmacokinetic features like PAINS filter, pharmacokinetic properties, pan assay interference, and interaction analysis. Finally, three phytochemicals, Withanone, Withametelin B, and Ixocarpanolide were identified as potential inhibitors for RfaH, having appreciable affinity of − 9.0, − 9.0 and − 8.8 kcal/mol specificity towards the binding pocket of RfaH. An all-atom molecular dynamic simulation was carried out for 500 ns to examine the structural flexibility and dynamic stability of RfaH and RfaH-ligand complexes, which revealed that complexes maintained stability throughout the given duration. All the selected compounds have shown drug-like properties as predicted from ADMET analysis and their physicochemical parameters. These compounds selectively bind to the crucial binding sites of RfaH and interact with important residues, preventing its binding with RNAP which can further be exploited as potential lead molecules against RfaH, providing a promising therapeutic avenue for combating antibiotic resistance.

Guava wilt is a devastating soil-borne disease that causes significant losses in guava orchards. Management of the disease is very challenging once established in the field. Therefore, there is a need to explore for an effective, economical, and sustainable management strategies. Aspergillus niger, a bio-control fungus, has been demonstrated effectiveness against various soil-borne pathogens including guava wilt pathogens. It produces a diverse hydrolysing enzymes and secondary metabolites. However, no extensive study has been undertaken to profile the secondary metabolites of A. niger. In this investigation, we assessed eleven A. niger strains (AN-1 to AN-11) against four guava wilt pathogens (Fusarium oxysporum f. sp. psidii, F. falciforme, F. chlamydosporum, and F. verticillioides) using a dual culture assay. All strains demonstrated effective by restricting the mycelial growth of pathogens, among them AN-11 displayed maximum inhibition of 86.33%, followed by the AN-3 (84.27%). The UPLC-QToF-ESIMS analysis was undertaken to explore the secondary metabolites of AN-11 responsible for inhibiting F. oxysporum f. sp. psidii. The crude extracts were obtained from F. oxysporum f. sp. psidii, AN-11 and their interaction using ethyl acetate as a solvent. After evaporating, the crude fractions were analysed using UPLC-QToF-ESIMS with an Acquity UPLC and a SCIEX SelexION Triple QuadTM 5500 System. From the ethyl acetate extract of F. oxysporum f. sp. psidii, approximately 14 metabolites involved in pathogenicity were identified. Similarly, analysis of AN-11 crude extract revealed 25 metabolites, and notably, 41 metabolites were identified during the interaction between AN-11 and F. oxysporum f. sp. psidii, including kotanin, isokotanin A, aurofusarin, kojic acid, pyranonigrin, aurasperone F, hexylitaconic acid, asperazine, bicoumanigrin, chloramphenicol, cephalosporin C, fusarin C, zearalonone, fonsecin B, malformin A, and others. Among these, 21 metabolites were produced only during the interaction and have antimicrobial properties. This study highlights the significant potential of the AN-11 strain in generating a diverse array of non-volatile secondary metabolites with antimicrobial properties. These metabolites could be further extracted and investigated for their efficacy against other soil borne pathogens and potentially developed into formulations for controlling plant diseases.

The increasing drug resistance of Mycobacterium tuberculosis (Mtb), coupled with the limited availability of effective anti-tuberculosis medications, poses significant challenges for the management and treatment of tuberculosis (TB). Globally, non-tuberculous mycobacteria (NTM) infections are increasing, with Mycobacterium avium complex and Mycobacterium abscessus (Mab) being the most common in labs and having few treatment options. There’s an urgent need for innovative therapies against Mtb and NTM that are effective and have minimal side effects. The study evaluated the in vitro efficacy of JIB-04, a Jumonji histone demethylase inhibitor, against Mtb, Mab, and multidrug-resistant (MDR) clinical isolates using the minimum inhibitory concentration (MIC) assay. We also determined the minimum bactericidal concentrations (MBCs) of JIB-04 against the H37Rv and H37Ra strains. A time-kill assay was performed to assess the comparative efficacy of JIB-04 and rifampicin against H37Ra. Additionally, the study investigated the impact of JIB-04 on biofilm formation and the persistence of H37Ra over extended periods. Our findings demonstrated a substantial inhibitory effect of JIB-04 on the growth of Mab, Mtb, and MDR clinical isolates. JIB-04 showed bactericidal effects at twice the MIC, outperforming rifampicin in reducing viable cell counts over 8 days. It showed moderate cytotoxicity to mammalian cells but effectively inhibited biofilm formation. In our anoxia model, JIB-04 induced a significant, concentration-dependent reduction in bacterial load. JIB-04 is a promising candidate for the treatment of MDR tuberculosis.

The respiratory bacterium nontypeable (non-encapsulated) Haemophilus influenzae (NTHi) is a key pathogen driving exacerbations in chronic obstructive pulmonary disease (COPD), and is associated with an excessive airway inflammation. Increasing issues with tolerance and unwanted side effects of existing pharmaceuticals present an urgent need for new, effective and minimally toxic therapeutic options. This study aimed to investigate the potential role of Tachyplesin III, an antimicrobial peptide derived from the hemolysates of Southeast Asian horseshoe crabs, in regulating NTHi-induced airway inflammation. The results revealed that Tachyplesin III effectively inhibited the production of IL-1β in NTHi-stimulated human lung epithelial cells (A549), without causing cytotoxic effects. Additionally, Tachyplesin III significantly reduced TNF-α, PGE₂ and NO production in NTHi-stimulated A549 cells. Moreover, this peptide inhibited the nuclear translocation of the NF-κB p65 subunit in NTHi-stimulated lung epithelial cells. It also reduced transcriptional activation of NF-κB target genes, as shown by lower mRNA levels of IL-1β, TNF-α, COX-2 and iNOS, which correlated with corresponding decreases in their protein expression. Tachyplesin III peptide also inhibited pro-IL-1β and NLRP3 protein expression and prevented NTHi-induced caspase-1 cleavage and IL-1β maturation. Together, our findings demonstrate that Tachyplesin III effectively reduced NTHi-mediated inflammation via the NF-κB/NLRP3 inflammasome signaling pathway, highlighting its important anti-inflammatory activity. Complementing these findings, in silico analysis revealed key pharmacokinetic and toxicological attributes, establishing a foundational understanding of Tachyplesin III as a promising therapeutic agent for managing NTHi-associated inflammation.

Penicillin-binding proteins (PBPs) are the targets of β-lactam antibiotics; however, changes in the affinity of PBPs for beta-lactam antibiotics often affect the susceptibility of bacteria to antibiotics. The purpose of this study was to elucidate the mechanism by which cefditoren, an oral third-generation cephalosporin, binds PBPs. The minimal inhibitory concentration (MIC), bactericidal curves, and inhibition zone comparisons were assessed to evaluate the antibacterial activity of cefditoren. PBP1A and PBP2X proteins from Streptococcus pneumoniae were purified, and their ability to bind to cefditoren was investigated via microscale thermophoresis. The Kd of cefditoren toward PBP1A was 0.005 ± 0.004 µM, which was lower than those of other cephalosporins (cefcapene, cefixime and cefdinir). In contrast, the Kd of cefditoren toward PBP2X of S. pneumoniae was 9.70 ± 8.24 µM, which was lower than that of cefixime but higher than those of cefcapene and cefdinir. Additionally, the biotinylated ampicillin (BIO-AMP) method was employed to evaluate the affinity of cefditoren toward PBPs of Haemophilus influenzae, and the results demonstrated that cefditoren and PBP3A/B had the lowest IC₅₀ values (0.060 ± 0.002 µM). These findings indicate that cefditoren has a strong affinity for PBP1A of H. influenzae. Cefditoren has a high affinity toward the PBP1As of S. pneumoniae and PBP1A and PBP3A/B of H. influenzae, which may contribute to the effective antibacterial effects of cefditoren against clinical strains and its low propensity for inducing resistance. The data presented in this article help elucidate the mechanism by which cefditoren, an oral third-generation cephalosporin, binds to PBPs and provide theoretical support for the wider use of cefditoren as an antibiotic therapy.

Pathogenic blue ear disease caused by porcine reproductive and respiratory syndrome virus (PRRSV) bring severe loss to breeding industry due to high infectivity and mortality. L. plantarum serves as the probiotic host strain, known for its beneficial properties in the gut microbiota. E. coli is used as a cloning host for the initial genetic engineering steps, facilitating the construction and amplification of the desired genetic constructs. In this study, using synthetic biology technology, we constructed engineered probiotics which could adhere and display nanobody on the surface to neutralize virus. Firstly, we screen an optimal nanobody to effectively bind with PRRSV by building library, expression and purification. Then, the integration of adhesion protein and nanobody into the genome of probiotics significantly improved its adhesion to IPEC-J2 cells. In addition, this engineered probiotic is almost non-toxic to cells with good safety, which can be used as a daily probiotics to prevent virus fecal transmission. Our study proposed this novel construction strategy of engineering probiotics with both adhesion and neutralization effects, which provided a new therapeutic view for intestinal virus clearance.

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