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ABSTRACTIn many Gram-positive bacteria, the transpeptidase enzyme sortase A (SrtA) anchors surface proteins to cell wall and plays a critical role in the bacterial pathogenesis. Here, we show that in Staphylococcus aureus, an important human pathogen, the SrtA is phosphorylated by serine/threonine protein kinase Stk1. S. aureus SrtA can also be phosphorylated by small-molecule phosphodonor acetyl phosphate (AcP) in vitro. We determined that various amino acid residues of S. aureus SrtA are subject to phosphorylation, primarily on its catalytic site residue cysteine-184 in the context of a bacterial cell lysate. Both Stk1 and AcP-mediated phosphorylation inhibited the enzyme activity of SrtA in vitro. Consequently, deletion of gene (i.e. stp1) encoding serine/threonine phosphatase Stp1, the corresponding phosphatase of Stk1, caused an increase in the phosphorylation level of SrtA. The stp1 deletion mutant mimicked the phenotypic traits of srtA deletion mutant (i.e. attenuated growth where either haemoglobin or haem as a sole iron source and reduced liver infections in a mouse model of systemic infection). Importantly, the phenotypic defects of the stp1 deletion mutant can be alleviated by overexpressing srtA. Taken together, our finding suggests that phosphorylation plays an important role in modulating the activity of SrtA in S. aureus.

The aim of this paper was to explore the process of β-glucosidase-assisted extraction of flavonoids from thinned fruits of Guanxi pummelo and the inhibitory effect of the extracted flavonoids on tyrosinase. In this paper, the extraction amount of total flavonoids was used as the index, and the β-glucosidase activity, enzymatic hydrolysis pH, temperature and time were optimized by single factor and response surface experiments, and the inhibitory effect of total flavonoids extract on tyrosinase was studied. The results showed that the optimal conditions for the extraction of flavonoids from thinned fruits of Guanxi pummelo were as follows: β-Glucosidase enzyme activity 0.0128 U/mL, enzymolysis pH4.4, temperature 35 ℃, time 4.8 h. Under these conditions, the extraction amount of total flavonoids was 34.0 mg/g, the half maximal inhibitory concentration (IC50) of the flavonoids extract on tyrosinase was 0.31 mg/mL. The results of this study showed that the enzymatic process could significantly improve the extraction amount of flavonoids from thinned fruits of Guanxi pummelo, and the flavonoids from thinned fruits of Guanxi pummelo had strong tyrosinase inhibition. The results of this study provided a reference for the extraction and application of flavonoids from thinned fruits of Guanxi pummelo, which was conducive to promoting the high-value utilization of Guanxi pummelo.

The focal point of probiotic efficacy and a crucial factor influencing poultry cultivation lies in the level of intestinal inflammation. In conventional farming processes, the reduction of intestinal inflammation generally proves advantageous for poultry growth. This study investigated the impact of Bacillus amyloliquefaciens TL (B.A.-TL) on inflammatory factor expression at both tissue and cellular levels, alongside an exploration of main active secondary metabolites. The results demonstrated that broiler feeding with a basal diet containing 4 × 109 CFU/kg B.A.-TL markedly enhanced chicken growth performance, concomitant with a significant decrease in the expression of genes encoding inflammatory cytokines (e.g., CCL4, CCR5, XCL1, IL-1β, IL-6, IL-8, LITAF, and LYZ) in jejunum and ileum tissues. The extracellular polysaccharides of B.A.-TL (EPS-TL) exhibited notable suppression of elevated inflammatory cytokine expression induced by Escherichia coli O55 lipopolysaccharides (LPS) in chicken macrophage-like cells (HD11) and primary chicken embryonic small intestinal epithelial cells (PCIECs). Moreover, EPS-TL demonstrated inhibitory effect on NF-κB signaling pathway activation. These findings suggested that the metabolic product of B.A.-TL (i.e., EPS-TL) could partly mitigate the enhanced expression of inflammatory factors induced by LPS stimulation, indicating its potential as a key component contributing to the anti-inflammatory effects of B.A.-TL.

ABSTRACT: The effects of pomegranate peel on the growth performance, intestinal morphology, and the cecal microbial community were investigated in broilers challenged with avian pathogenic Escherichia coli (APEC) O78. A total of 240 one-day-old chicks (120 males and 120 females) were randomly and evenly allotted into 4 treatment groups (each with 6 biological replicates each of 10 chicks), i.e., negative control (NC), positive control (PC), and 2 experimental groups treated with 0.2% fermented pomegranate peel (FP) and 0.2% unfermented pomegranate peel (UFP), respectively, with PC, FP, and UFP groups challenged with APEC O78 (5 × 108 CFU) on day 14. Results showed that the challenge of APEC O78 decreased the body weight (BW) and average daily gain (ADG) of broilers from 1 to 28 d (P < 0.01). These broilers exhibited more pathological conditions in the heart and liver and higher mortality rates in 28 d compared to the NC group. Diet supplemented with pomegranate peel (either fermented or unfermented) significantly increased BW, ADG, and the villus height/crypt depth ratio (VCR) of small intestine in 28 d compared to the NC group (P < 0.05). Results of the taxonomic structure of the gut microbiota showed that compared to the NC group, the APEC challenge significantly decreased the relative abundance of Bacteroidetes and increased the relative abundance of Firmicutes (P < 0.01). Compared to the PC group, the relative abundance of Ruminococcus_torques_group in FP group was increased, while the relative abundance of Alistipes was decreased. In summary, our study showed that the dietary supplementation of pomegranate peel could maintain the intestinal microbiota at a state favorable to the host, effectively reduce the abnormal changes in the taxonomic structure of the intestinal microbiota, and improve the growth performance in broilers treated with APEC.

Background: The use of single cell protein (SCP) has become a method for alleviating the shortage of protein feed that microorganisms propagate in a suitable culture medium. In this study, SCP was produced by yeast to use the nutrition contained in the biogas slurry of chicken manure. Results: The results showed that Galactomyces candidum was the most efficient at producing SCP among the seven yeasts studied. The maximum cell dry weight (CDW) 6.79 g/L and protein content 39.39%, were obtained under the fermentation conditions of initial NH4+-N concentration of 2000 mg/L and a C/N ratio of 6:1 with acetate as the pH regulator. The total CDW increased to 9.24 g/L after secondary fermentation. Metal elements had a little effect on the growth of G. candidum. The addition of sulfur not only promoted the synthesis of sulfur-containing amino acid cysteine but also increased protein content by promoting the synthesis of glutamic acid and glutamine. Conclusions: Future experiments should focus more on achieving high-density cultivation and more efficient utilization of ammonia nitrogen in the biogas slurry.How to cite: Zhou Pan, Zhang Lu, Ding Hongxia, et al. Optimization of culture conditions of screened Galactomyces candidum for the production of single cell protein from biogas slurry. Electron J Biotechnol 2021;55. https://doi.org/10.1016/j.ejbt.2021.11.006

Atrial fibrillation (AF), the most frequent cause of cardioembolic stroke, is increasing in prevalence as the population ages, and presents with a broad spectrum of symptoms and severity. The early identification of AF is an essential part for preventing the possibility of blood clotting and stroke. In this work, a real-time algorithm is proposed for accurately screening AF episodes in electrocardiograms. This method adopts heart rate sequence, and it involves the application of symbolic dynamics and Shannon entropy. Using novel recursive algorithms, a low-computational complexity can be obtained. Four publicly-accessible sets of clinical data (Long-Term AF, MIT-BIH AF, MIT-BIH Arrhythmia, and MIT-BIH Normal Sinus Rhythm Databases) were used for assessment. The first database was selected as a training set; the receiver operating characteristic (ROC) curve was performed, and the best performance was achieved at the threshold of 0.639: the sensitivity (Se), specificity (Sp), positive predictive value (PPV) and overall accuracy (ACC) were 96.14%, 95.73%, 97.03% and 95.97%, respectively. The other three databases were used for independent testing. Using the obtained decision-making threshold (i.e., 0.639), for the second set, the obtained parameters were 97.37%, 98.44%, 97.89% and 97.99%, respectively; for the third database, these parameters were 97.83%, 87.41%, 47.67% and 88.51%, respectively; the Sp was 99.68% for the fourth set. The latest methods were also employed for comparison. Collectively, results presented in this study indicate that the combination of symbolic dynamics and Shannon entropy yields a potent AF detector, and suggest this method could be of practical use in both clinical and out-of-clinical settings.