Development of a Chemically Defined Medium for Better Yield and Purification of Enterocin Y31 from Enterococcus faecium

The macro- and micronutrients in traditional medium, such as MRS, used for cultivating lactic acid bacteria, especially for bacteriocin production, have not been defined, preventing the quantitative monitoring of metabolic flux during bacteriocin biosynthesis. To enhance Enterocin Y31 production and simplify steps of separation and purification, we developed a simplified chemically defined medium (SDM) for the growth of Enterococcus faecium Y31 and production of its bacteriocin, Enterocin Y31. We found that the bacterial growth was unrelated to Enterocin Y31 production in MRS; therefore, both the growth rate and the Enterocin Y31 production were set as the index for investigation. Single omission experiments revealed that 5 g/L NaCl, five vitamins, two nucleic acid bases, MgSO4·7H2O, MnSO4·4H2O, KH2PO4, K2HPO4, CH3COONa, fourteen amino acids, and glucose were essential for the strain’s growth and Enterocin Y31 production. Thus, a novel simplified and defined medium (SDM) was formulated with 30 components in total. Consequently, Enterocin Y31 production yield was higher in SDM as compared to either MRS or CDM. SDM improved the Enterocin Y31 production and simplified the steps of purification (only two steps), which has broad potential applications

Purification and Partial Characterization of Bacteriocin Lac-B23, a Novel Bacteriocin Production by Lactobacillus plantarum J23, Isolated From Chinese Traditional Fermented Milk

The exploration and evaluation of bacteriocin-producing lactic acid bacteria (LAB) have been one of the powerful means to food preservation. A total of 300 strains were isolated from Chinese traditional fermented milk products. A bacteriocin-producing LAB, named Lactobacillus plantarum J23, was screened and identified. Bacteriocin Lac-B23 from L. plantarum J23 was purified by 80% ammonium sulfate precipitation, cation-exchange chromatography, and reverse-phase high-performance liquid chromatography. Molecular weight of bacteriocin Lac-B23 was determined to be approximately 6.73 kDa by tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, and it was confirmed as a novel bacteriocin by liquid chromatography-mass spectrometry. Moreover, bacteriocin Lac-B23 showed thermal stability when heated at below 100°C for 30 min, pH stability between pH 2.0 and 12.0, and sensitivity to trypsin, proteinase K, and proteinase E. The antimicrobial activity of bacteriocin could be enhanced by addition of Fe2+, Mn2+, and ethyl alcohol, and inhibited by Cu2+, K+, Ca2+, Zn2+, Mg2+, and sodium chloride. The results suggested bacteriocin Lac-B23 to have potential application prospects in the food industry

Enhancement of Innate Immune Function in Mice by Bifidobacterium bifidum FL-228.1

In this study, eight potential functional strains were selected to interfered with RAW264.7 murine macrophages and human peripheral blood mononuclear cells (PBMCs). Then, changes in phagocytic activity of RAW264.7 cells and natural killer (NK) cell activity were detected and the screened potential probiotics were further intervened in BALB/c mice to explore their immunomodulatory efficacy in vivo. In cell experiments, the results showed that the intervention of different strains significantly increased the phagocytic activity of RAW264.7 cells (P0.05). In conclusion, Bifidobacterium bifidum FL-228.1 can improve innate immune function and have a more comprehensive effect on the immune system by regulating immune cell activity, cytokine expression and mRNA levels of immune molecules related to antimicrobial peptides

The deubiquitinase USP6 affects memory and synaptic plasticity through modulating NMDA receptor stability

人类与其他动物相比的重要区别在于人类拥有高等认知能力，这种能力集中体现在学习记忆和语言表达方面。厦门大学医学院神经科学研究所王鑫教授团队发现人科动物特异性基因USP6作为一个新的NMDA受体调控因子，可通过去泛素化途径调节NMDA型谷氨酸受体的降解和稳定性，进而调控突触可塑性和学习记忆能力。 本研究工作由王鑫教授指导完成，博士生曾凡伟、马学海与硕士生朱琳为共同第一作者，王鑫教授为通讯作者。Ubiquitin-specific protease (USP) 6 is a hominoid deubiquitinating enzyme previously implicated in intellectual disability and autism spectrum disorder. Although these findings link USP6 to higher brain function, potential roles for USP6 in cognition have not been investigated. Here, we report that USP6 is highly expressed in induced human neurons and that neuron-specific expression of USP6 enhances learning and memory in a transgenic mouse model. Similarly, USP6 expression regulates N-methyl-D-aspartate-type glutamate receptor (NMDAR)-dependent long-term potentiation and long-term depression in USP6 transgenic mouse hippocampi. Proteomic characterization of transgenic USP6 mouse cortex reveals attenuated NMDAR ubiquitination, with concomitant elevation in NMDAR expression, stability, and cell surface distribution with USP6 overexpression. USP6 positively modulates GluN1 expression in transfected cells, and USP6 down-regulation impedes focal GluN1 distribution at postsynaptic densities and impairs synaptic function in neurons derived from human embryonic stem cells. Together, these results indicate that USP6 enhances NMDAR stability to promote synaptic function and cognition.This work was partially supported by the National Natural Science Foundation of China (31871077, 81822014, 81571176 to XW; 81701349 to Hongfeng Z.; 81701130 to QZ; and 81471160 to HS), the National Key R&D Program of China (2016YFC1305900 to XW and HS), the Natural Science Foundation of Fujian Province of China (2017J06021 to XW), the Fundamental Research Funds for the Chinese Central Universities (20720150061 to XW and 20720180040 to ZS), Open Research Fund of State Key Laboratory of Cellular Stress Biology, Xiamen University (SKLCSB2019KF012 to QZ), and China Postdoctoral Science Foundation (2017M612130 to QZ).该研究得到了国家自然科学基金面上项目和优秀青年基金项目的支持

Prospects of Artificial Kefir Grains Prepared by Cheese and Encapsulated Vectors to Mimic Natural Kefir Grains

Kefir is a natural fermented dairy beverage obtained by fermenting milk with kefir starter grains. However, up to now, there is still no efficient approach to producing stable kefir grains by using the pure or cultural mixture of strains isolated from the original kefir grains. Therefore, new techniques need to be taken to promote the kefir grain production. To this purpose, an encapsulated vector produced by entrapment of the dominant strains isolated from kefir grain and the cheese vector which was produced by a traditional manufacturing method was used to mimic kefir grain forming, respectively. Then, the composition, microstructure, and microflora of the two vectors were investigated and were compared with the natural kefir grains. Results indicated that the protein and polysaccharide content of cheese vector were much higher than encapsulated vector; the distribution of microorganisms inside the cheese vector was more similar to that inside the natural kefirs. It indicated that the cheese vector would be more suitable to mimic kefir grain production. Results of the present investigations reveal the potential of the cheese vector for kefir grains production at the industrial level

<i>Lactobacillus paracasei</i> subsp. <i>paracasei</i> X12 Strain Induces Apoptosis in HT-29 Cells through Activation of the Mitochondrial Pathway

L. paracasei subsp. paracasei X12 was obtained from traditional cheese produced in northwestern China. In this study, we showed that whole peptidoglycan (WPG), extracted from L. paracasei subsp. paracasei X12, inhibited proliferation and induced apoptosis in HT-29 cells in a dose-dependent manner. In addition, WPG-induced apoptosis was associated with the loss of mitochondrial membrane potential (Ψm), the release of cytochrome c (Cyto-C) from mitochondrialto cytosolic spaces, activation of Caspase 3, and accumulation of intracellular reactive oxygen species (ROS). Finally, semi-quantitative RT-PCR showed that these events were accompanied by upregulation of proapoptotic genes (Bax or Bad) and downregulation of antiapoptotic genes (Bcl-xl). Taken together, our results demonstrated that WPG induced apoptosis in HT-29 cells through activation of the mitochondrial pathway. WPG exerted only minor toxicity upon noncancerous cells and therefore might be used as a natural agent in the treatment of cancer in future

Physical, textural, and rheological properties of whipped cream affected by milk fat globule membrane protein

This work aims at improving the textural and whipping properties of whipped cream by the addition of milk fat globule membrane protein. The determination of particle size distribution and average diameter of whipped cream showed that the small particle size was shifted to a larger range after milk fat globule membrane protein was added. The average particle size (d3,2) of whipped cream reached a maximum value of 5.05 µm at 1% milk fat globule membrane protein, while slowly decreased with increasing milk fat globule membrane protein levels from 2% to 5%. In addition, the partial coalescence of fat increased with the increase of milk fat globule membrane protein levels, and the correlation between the whipping time and the overrun of whipped cream was positive. The addition of milk fat globule membrane protein also altered the rheological behaviour of whipped cream, resulting in the increase of modulus G′ and the loss modulus G″. The results also indicated that higher milk fat globule membrane protein level decreased the serum loss of whipped cream while improved its stability. While milk fat globule membrane protein levels had no significant effect on viscosity, its increasing levels effectively improved the hardness, consistency, and viscosity of whipped cream

Improvement of the Texture of Yogurt by Use of Exopolysaccharide Producing Lactic Acid Bacteria

19 Streptococcus thermophilus with high exopolysaccharide production were isolated from traditional Chinese fermented dairy products. The exopolysaccharide and viscosity of milk fermented by these 19 isolates were assayed. The strains of Streptococcus thermophilus zlw TM11 were selected because its fermented milk had the highest exopolysaccharide content (380 mg/L) and viscosity (7716 mpa/s). Then Streptococcus thermophilus zlw TM11 was combined with Lactobacillus delbrueckii subsp. bulgaricus 3 4.5 and the combination was named SH-1. The quality of the yogurt fermented by SH-1 and two commercial starter cultures (YO-MIX 465, YF-L711) were compared. It was shown that the exopolysaccharide content of yogurt fermented by SH-1 was similar to that of yogurt fermented by YF-L711 and significantly higher than YO-MIX 465 (p<0.05). In addition, the yogurt fermented by SH-1 had the lowest syneresis (8.5%) and better texture and sensory than the samples fermented by YO-MIX 465 and YF-L711. It manifested that the selected higher exopolysaccharide production starter SH-1 could be used as yogurt starter and reduce the amount of adding stabilizer, which can compare with the imported commercial starter culture

Application of plantaricin Q7 in the storage and preservation of milk

Plantaricin Q7 is a bacteriocin produced by Lactobacillus plantarum Q7 with broad-spectrum antibacterial activity.It has the advantages of safety,high efficiency and good stability.In this study,plantaricin Q7 with a titer of 533 AU·mL-1 was added to milk to explore its preservation effects in the process of milk storage.The result showed that plantaricin Q7 maintained the good sense of milk.There was no peculiar smell on the 13th day of storage,and the milk remained white and slightly yellow,without stratification and precipitation.Pseudomonas fluorescens was the dominant psychrophilic spoilage bacteria in milk.Without adding plantaricin Q7,the number of P.fluorescens in milk exceeded the national standard on the 7th day.After adding plantaricin Q7,the time of P. fluorescens exceeding the national standard was postponed for 4 days.Plantaricin Q7 effectively slowed down the increase of milk acidity and the decrease of milk pH value.The results provided a basis for the development and application of plantaricin Q7

Physical activity and fasting glucose in adults with abnormal glucose metabolism: Findings from two independent cross-sectional studies in China

Abstract Background: Relationship between physical activity and fasting glucose in people with abnormal glucose metabolism is not well-known. This study was to investigate dose-response association between physical activity (PAT) and fasting glucose from two independent surveys among Chinese adults with abnormal glucose metabolism. Methods: 9419 adults with abnormal glucose metabolism from two independent surveys among Chinese adults were analyzed. Demographics, level of fasting glucose and PAT (in Met Score) were measured. Dose-response relationship between fasting glucose and PAT was assessed by natural cubic spline model. Certain threshold point was identified, and linear regression models were then used within each threshold interval to assess the liner relationship functions. Models were adjusted for confounding factors and were stratified in subgroup analyses by the main population characteristics including survey site, gender and age-group. Results: Overall the relationship between PAT and fasting glucose was not in a linear association (Linearity test: p < 0.0001). Level of fasting glucose was not associated with amount of PAT until a threshold point (square-rooted Met Score 66.6 (original Met score: 4436 MET-minutes per week), 95% confidence intervals (65.2-69.3 (4,251-4,802 MET-minutes per week)). After this threshold, an inverse association was observed: each increase of every standard deviation of square-rooted Met Score 29.8 (888 MET-minutes per week) was associated with a 0.25 mmol/L decrease in fasting glucose, with adjustment for confounding factors. The patterns of relationship were tested to be consistent in subgroup analyses by survey site, gender and age group. Conclusions: Our study indicated that among adults with abnormal glucose metabolism the level of fasting glucose was only inversely associated with square-rooted Met Score beyond a certain square-rooted Met Score amount