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1.
Cancer Cell Int ; 21(1): 70, 2021 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-33482814

RESUMEN

BACKGROUND: Gastric cancer is one of the most prevalent and deadly malignancies without efficient treatment option. This study aimed to investigate the effect of hydrogen gas on the behavior of gastric cancer cells. METHODS: Gastric cancer cell lines MGC-803 and BGC-823 were treated with or without H2 /O2 gas mixture (66.7%:33.3% v/v). Proliferation and migration were assessed by MTT and scratch wound healing assays respectively. The expression of lncRNA MALAT1, miR-124-3p, and EZH2 was analyzed by real-time quantitative PCR and/or western blot. Tumor growth was estimated using xenograft mouse model. RESULTS: H2 gas significantly inhibited gastric tumor growth in vivo and the proliferation, migration, and lncRNA MALAT1 and EZH2 expression of gastric cancer cells while upregulated miR-124-3p expression. LncRNA MALAT1 overexpression abolished all the aforementioned effects of H2. LncRNA MALAT1 and miR-124-3p reciprocally inhibited the expression of each other. MiR-124-3p mimics abrogated lncRNA MALAT1 promoted EZH2 expression and gastric cancer cell proliferation and migration. CONCLUSIONS: These data demonstrated that H2 might be developed as a therapeutics of gastric cancer and lncRNA MALAT1/miR-124-3p/EZH2 axis could be a target for intervention.

2.
Org Biomol Chem ; 19(36): 7914-7919, 2021 Sep 22.
Artículo en Inglés | MEDLINE | ID: mdl-34549232

RESUMEN

A new two-step one-pot aminobromination/chlorination of carbonyl alkynes has been achieved via a Michael addition of aliphatic secondary amines and subsequent ß-bromination/chlorination of the obtained enamines to afford various α-X (X = Br or Cl) enamino ketones/esters in moderate to good yields. A solvent-controllable protocol has been developed to produce versatile 3-(2,5-dioxopyrrolidin-1-yl)acrylates in moderate yields by using toluene as the solvent and chain alkyl propiolates as alkynyl substrates.

3.
Microb Cell Fact ; 19(1): 214, 2020 Nov 23.
Artículo en Inglés | MEDLINE | ID: mdl-33228718

RESUMEN

BACKGROUND: Wheat sheath blight, a soil borne fungal disease caused by Rhizoctonia cerealis, is considered as one of the most serious threats to wheat worldwide. Bacillus subtilis Z-14 was isolated from soil sampled from a wheat rhizosphere and was confirmed to have strong antifungal activity against R. cerealis. RESULTS: An antifungal protein, termed F2, was isolated from the culture supernatant of Z-14 strain using precipitation with ammonium sulfate, anion exchange chromatography, and reverse phase chromatography. Purified F2 had a molecular mass of approximately 8 kDa, as assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Edman degradation was used to determine the amino acid sequence of the N-terminus, which was NH2ASGGTVGIYGANMRS. This sequence is identical to a hypothetical protein RBAM_004680 (YP_001420098.1) synthesized by B. amyloliquefaciens FZB42. The recombinant F2 protein (rF2) was heterologously expressed in the yeast host Pichia pastoris, purified using a Niaffinity column, and demonstrated significant antifungal activity against R. cerealis. The purified rF2 demonstrated broad spectrum antifungal activity against different varieties of fungi such as Fusarium oxysporum, Verticillium dahliae, Bipolaris papendorfii, and Fusarium proliferatum. rF2 was thermostable, retaining 91.5% of its activity when incubated for 30 min at 100 °C. Meanwhile, rF2 maintained its activity under treatment by proteinase K and trypsin and over a wide pH range from 5 to 10. CONCLUSIONS: A novel antifungal protein, F2, was purified from biocontrol Bacillus subtilis Z-14 strain fermentation supernatant and heterologously expressed in Pichia pastoris to verify its antifungal activity against R. cerealis and the validity of the gene encoding F2. Considering its significant antifungal activity and stable characteristics, protein F2 presents an alternative compound to resist fungal infections caused by R. cerealis.


Asunto(s)
Hongos/efectos de los fármacos , Fungicidas Industriales/farmacología , Enfermedades de las Plantas/prevención & control , Secuencia de Aminoácidos , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Agentes de Control Biológico/farmacología , Pruebas de Sensibilidad Microbiana , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/farmacología , Rizosfera , Saccharomycetales/metabolismo , Suelo , Microbiología del Suelo , Triticum/microbiología
4.
Opt Lett ; 39(11): 3212-5, 2014 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-24876015

RESUMEN

Optical bistability typically occurs only when the optical thickness in the device or the input light power is unfavorably large. Here we show that, for a class of plasmonic metamaterials consisting of ultrathin holey metallic plates filled with nonlinear materials, the optical bistability can occur with an ultralow excitation power. We present a realistic design working at 0.2 THz and perform full-wave simulations to quantitatively study its optical bistability properties. An analytical model is developed to explain the inherent physics and provides a general design guideline for future development.

5.
Zhonghua Yu Fang Yi Xue Za Zhi ; 48(9): 771-5, 2014 Sep.
Artículo en Zh | MEDLINE | ID: mdl-25492287

RESUMEN

OBJECTIVE: To describe the prevalence and relationship between screen time and sub-health status among middle school students in weekdays and weekends. METHODS: With multistage stratified random cluster sampling, 14 400 students from junior and senior middle schools in 288 classes of 30 schools in 4 cities( Shenyang, Chongqing, Xinxiang, Guangzhou) were investigated in October, 2011. A total of 13 817 valid questionnaires were retained for analysis. Survey content includes general demographic information, screen time, sub-health state. Logistic regression model was used to analyze the relationship between screen time and sub-health status. RESULTS: The screen time among middle school students > 2 h/d on weekdays and weekends were 15.1% (2 085/13 817) and 58.5% (8 077/13 817) , respectively. Among them, the boys' screen time > 2 h/d on weekdays and weekends were 17.9% (1 187/6 644) and 61.3% (4 070/6 644), which were higher than girls (12.5% (898/7 173), 55.9% (4 007/7 173) )(χ² values were 76.95 and 41.35, respectively. P < 0.01). The prevalence of physical sub-health status, psychological sub-health status and sub-health status were 24.8% (3 431/13 817), 21.7% (2 997/13 817) and 22.1% (3 051/13 817), respectively. Among them, the girls' prevalence of physical sub-health status, psychological sub-health status and sub-health status were 26.0% (1 863/7 173), 22.9% (1642/7 173) and 23.1% (1 658/7 173) ), respectively, which were higher than boys' (23.6% (1 568/6 644), 20.4% (1 355/6 644) and 21.0% (1 393/6 644)) (χ² values were 10.39, 12.66 and 9.25, respectively. P < 0.05). The senior students' prevalence of physical sub-health status, psychological sub-health status and sub-health status were 31.6% (2 176/6 884) , 28.3% (1 946/6 884) and 28.9% (1 990/6 884), respectively, which were higher than junior students' (18.1% (1 255/6 933), 15.2% (1 051/6 933) and 15.3% (1 061/6 933)) (χ² values were 337.65, 349.46 and 371.54, respectively, P < 0.05). After adjusting on demographic characteristics, compared with screen time ≤ 2 h/d on weekday, screen time > 2 h/d on weekday was significantly associated with physical sub-health status, psychological sub-health status and sub-health status (OR (95% CI): 1.39 (1.25-1.55), 1.62 (1.45-1.81) and 1.59 (1.43-1.78)). And compared with screen time ≤ 2 h/d on weekend, screen time > 2 h/d on weekend was also significantly associated with physical sub-health status, psychological sub-health status and sub-health status (OR(95% CI):1.49 (1.37-1.61), 1.81 (1.66-1.98), and 1.71 (1.57-1.87)). CONCLUSION: The rate of the screen time > 2 h/d and the detection ratio of sub-health among the middle school students in Shenyang, Chongqing, Xinxiang, Guangzhou in 2011 were high. Screen time >2 h/d on weekdays or weekends was a risk factor for sub-health status.


Asunto(s)
Estado de Salud , Microcomputadores/estadística & datos numéricos , Televisión/estadística & datos numéricos , Adolescente , China/epidemiología , Ciudades , Recolección de Datos , Femenino , Humanos , Modelos Logísticos , Masculino , Prevalencia , Factores de Riesgo , Instituciones Académicas , Factores Sexuales , Estudiantes , Encuestas y Cuestionarios , Población Urbana/estadística & datos numéricos
6.
Int J Biol Macromol ; 253(Pt 5): 127059, 2023 Dec 31.
Artículo en Inglés | MEDLINE | ID: mdl-37769756

RESUMEN

The absolute amount of nutrients on plant leaves is usually low, and the growth of epiphytic bacteria is typically limited by nutrient content. Thus, is of great significance to study the survival mechanism of epiphytes under nutritional stress for plant disease control. In this paper, Pantoea agglomerans CHTF15 isolated from walnut leaves was used to detect the key genes for the survival of the bacterium under simulated nutrient stress in artificial medium. Genome sequencing was combined with transposon insertion sequencing (Tn-seq) for the detection technique. A total of 105 essential genes were screened from the whole genome. The genes were mainly related to the nucleotide metabolism, protein metabolism, biological oxidation and the gene repair of bacteria analyzed by gene ontology (GO) enrichment analysis. Volcano map analysis demonstrated that the functions of the 15 genes with the most significant differences were generally related to the synthesis of amino acids or proteins, the nucleotide metabolism and homologous recombination and repair. Competitive index analysis revealed that the deletion of the genes dksA and epmA regulating protein synthesis, the gene ribB involved in the nucleotide metabolism and the gene xerD involved in recombination repair induced a significant reduction in the survival ability of the corresponding mutants in the 0.10 % YEP medium and the walnut leaf surface. The results act as a foundation for further in-depth research on the infection process and the mechanisms of pathogenic bacteria.


Asunto(s)
Pantoea , Pantoea/genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Secuencia de Bases , Nucleótidos/metabolismo
7.
J Agric Food Chem ; 71(1): 197-210, 2023 Jan 11.
Artículo en Inglés | MEDLINE | ID: mdl-36573896

RESUMEN

Water-soluble humic materials (WSHMs) can enhance the nodule numbers of soybean plants. In this study, targeted metabolomics and transcriptomics were used to understand this mechanism. Results showed that 500 mg/L WSHM increased the adsorption and colonization of rhizobia in soybean roots. High-performance liquid chromatography and targeted metabolomics showed that WSHMs could regulate the content and distribution of endogenous hormones of soybean plants at the initial stage of soybean nodulation. Transcriptomic analysis showed a total of 2406 differentially expressed genes (DEGs) by the 25th day, accounting for 4.89% of total annotation genes (49159). These DEGs were found to contribute primarily to the MAPK signaling pathway, glycolysis/gluconeogenesis, and plant hormone signal transduction according to the -log 10 (Padjust) value in the KEGG pathway. Subsequently, DEGs related to these hormones were selected for verification using quantity-PCR. The WSHM increased the number of nodules by regulating the expression of endogenous hormones in soybean plants.


Asunto(s)
Glycine max , Transcriptoma , Glycine max/metabolismo , Agua/metabolismo , Proteínas de Plantas/metabolismo , Metabolómica , Hormonas/metabolismo , Regulación de la Expresión Génica de las Plantas , Nodulación de la Raíz de la Planta/genética
8.
Transl Cancer Res ; 12(12): 3443-3452, 2023 Dec 31.
Artículo en Inglés | MEDLINE | ID: mdl-38192976

RESUMEN

Background: Rho guanine nucleotide exchange factor 16 (ARHGEF16) is a newly discovered Rho-family guanine nucleotide exchange factor (GEF) involved in the activation of Rho-family GTPases. However, its roles in colon cancer cell proliferation, migration, and invasion remain unknown. This study analyzed the expression of ARHGEF16 in colon cancer and explored its biological effects on colon cancer cells, so as to find new therapeutic targets for the treatment of colon cancer. Methods: The expression of ARHGEF16 in colon cancer tissues and cells was detected by bioinformatics analysis, western blot, and quantitative real time polymerase chain reaction (qRT-PCR) assays. The effects of overexpression and silencing of ARHGEF16 on the biological behavior of colon cancer cells were examined by Cell Counting Kit-8 (CCK-8), cell scratching and Transwell assays. Results: The database showed that ARHGEF16 was highly expressed in colon cancer tissues. Validation with clinical fresh tissue specimens and colon cancer cell lines revealed that ARHGEF16 was highly expressed in both. The proliferation, growth, migration, and invasion ability of colon cancer cell lines increased significantly with the overexpression of ARHGEF16, while silencing ARHGEF16 showed the opposite effect. Conclusions: The expression of ARHGEF16 is closely related to the migration and invasive ability of colon cancer cells, and overexpression of ARHGEF16 promotes the migration and invasion of colon cancer cells and correlates with the metastatic potential of colon cancer.

9.
J Agric Food Chem ; 71(5): 2644-2657, 2023 Feb 08.
Artículo en Inglés | MEDLINE | ID: mdl-36706360

RESUMEN

Controlling cucumber Fusarium wilt caused by Fusarium oxysporum f. sp. cucumerinum (FOC) with Bacillus strains is a hot research topic. However, the molecular mechanism of Bacillus underlying the biocontrol of cucumber wilt is rarely reported. In this study, B. subtilis strain Z-14 showed significant antagonistic activity against FOC, and the control effect reached 88.46% via pot experiment. Microscopic observations showed that strain Z-14 induced the expansion and breakage of FOC hyphae. The cell wall thickness was uneven, and the organelle structure was degraded. The combined analysis of metabolome and transcriptome showed that strain Z-14 inhibited the FOC infection by inhibiting the synthesis of cell wall and cell membrane, energy metabolism, and amino acid synthesis of FOC mycelium, inhibiting the clearance of reactive oxygen species (ROS) and the secretion of cell wall-degrading enzymes (CWDEs), thereby affecting mitogen-activated protein kinase (MAPK) signal transduction and inhibiting the transport function.


Asunto(s)
Bacillus , Cucumis sativus , Fusarium , Bacillus subtilis , Enfermedades de las Plantas , Perfilación de la Expresión Génica
10.
Front Plant Sci ; 14: 1196939, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37564385

RESUMEN

Introduction: Nitrogen fertilizer intake promotes soybean growth before the formation of nodules, but excess nitrogen has an inhibitory effect on soybean nodulation. It is important to balance nitrogen levels to meet both growth and nodulation needs. Methods: the nitrogen level suitable for soybean growth and nodulation was studied, the role of humic acid (HA) in alleviating the inhibition of high nitrogen on soybean nodulation was analyzed, and transcriptomic analysis was performed to understand its mechanism. Results: The results showed that a lower level of nitrogen with 36.4 mg urea per pot could increase the number of nodules of soybean, and a higher level of nitrogen with 145.9 mg urea per pot (U4 group) had the best growth indicators but inhibited nodulation significantly. HA relieved the inhibitory effect at high nitrogen level, and the number of nodules increased by 122.1% when 1.29 g HA was added (H2 group) compared with the U4 group. The transcriptome analysis was subsequently performed on the H2 and U4 groups, showing that there were 2995 differentially expressed genes (DEGs) on the 25th day, accounting for 6.678% of the total annotated genes (44,848) under the test conditions. These DEGs were enriched in mitogen-activated protein kinase signaling pathway-plant, flavonoid biosynthesis, and plant hormone signal transduction based on the -log10 (P adjusted) value in the Kyoto Encyclopedia of Genes and Genomes pathway (KEGG). Discussion: HA balanced the nitrogen level through the above pathways in soybean planting to control the number of nodules.

11.
Front Microbiol ; 14: 1330538, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38264477

RESUMEN

Whole-plant corn silage (WPCS) has been widely used as the main roughage for ruminant, which promoted the utilization of corn stover for animal feed production. However, rigid cell wall structure of corn stover limits the fiber digestion and nutrients adsorption of WPCS. This study investigated the effect of adding cellulase on improving the fermentation quality of WPCS ensiling with a Bacillus complex inoculant. With the Bacillus (BA), the lactic acid accumulation in the WPCS was significantly higher than that in control (CK). The additive cellulase (BC) increased the lactic acid content to the highest of 8.2% DW at 60 days, which was significantly higher than that in the CK and BA groups, and it reduced the neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents from 42.5 to 31.7% DW and 28.4 to 20.3% DW, respectively, which were significantly lower than that in the CK and BA groups. The crude protein and starch were not obviously lost. Dynamic microbial community analysis showed that the Bacillus inoculant promoted the lactic acid bacteria (LAB) fermentation, because higher abundance of Lactobacillus as the dominant bacteria was observed in BA group. Although the addition of cellulase slowed the Lactobacillus fermentation, it increased the bacterial community, where potential lignocellulolytic microorganisms and more functional enzymes were observed, thus leading to the significant degradation of NDF and ADF. The results revealed the mechanism behind the degradation of NDF and ADF in corn stover, and also suggested the potential of cellulase for improving the nutritional quality of WPCS.

12.
Front Nutr ; 9: 1017637, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36570163

RESUMEN

Microbial fermentation is an effective method to degrade free-gossypol, which is a toxic substance restricting the utilization of cottonseed meal in animal husbandry. However, there are few researches on the nutritional effect and the change of bacterial community on cottonseed meal fermented with anaerobic solid-state fermentation. This study evaluated the effects of fermentation with Bacillus sp. on gossypol degradation and nutritional quality improvement in cottonseed meal (CM), as well as the changes of bacterial community structure during fermentation. The strains with high activity for digesting free gossypol were screened from high protease-producing strains preserved in the laboratory. Then the strains which had both the gossypol degradation activity and protease producing activity were selected to degrade macromolecular protein and free gossypol in CM. The unsterilized SSF medium was inoculated with 109 CFU/kg Bacillus culture and fermented at room temperature for 14 days. Each group had three parallels. And the effects of anaerobic solid-state fermentation on unsterilized CM was evaluated. Results showed that for the seven strains with high activity for digesting free gossypol and producing protease that were screened, free gossypol content in fermented cottonseed meal (FCM) decreased and acid-soluble protein (ASP) contents increased. Among them, strain M-15 had the best fermentation effect, with the free gossypol degradation rate of 93.46% and acid soluble protein content of 13.26%. M-15 was identified as Bacillus subtilis. During fermentation with M-15, the bacterial diversity in CM was reduced, but not significant and the community structure was simpler significantly. The strain M-15 selected in this experiment reduced the free gossypol content and improved the nutritional quality of CM through anaerobic solid-state fermentation, which can be used for industrial large-scale production.

13.
Front Chem ; 10: 951649, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35991596

RESUMEN

The chromatographic column is the core of a high-performance liquid chromatography (HPLC) system, and must have excellent separation efficiency and selectivity. Therefore, functional modification materials for monolithic columns have been rapidly developed. This study is a systematic review of the recently reported functionalized monolithic columns. In particular, the study reviews the types of functional monomers under different modification conditions, as well as the separation and detection techniques combined with chromatography, and their development prospects. In addition, the applications of functionalized monolithic columns in food analysis, biomedicine, and the analysis of active ingredient of Chinese herbal medicines in recent years are also discussed. Also reviewed are the functionalized monolithic columns for qualitative and quantitative analysis. It provided a reference for further development and application of organic polymer monolithic columns.

14.
Artículo en Inglés | MEDLINE | ID: mdl-35662876

RESUMEN

Graphene oxide (GO) was introduced into a monolithic column and a network porous poly (GO-co-TAIC-co-MMA) monolith was prepared by redox polymerization. The internal morphology and pore size distribution of the polymer were observed by scanning electron microscopy, and the nitrogen adsorption-desorption and mercury intrusion methods. After optimization, 8 kinds of aromatic compounds were effectively separated in 5 min, and the theoretical plates number of the monolithic column exceeded 33, 070 plates m-1. Five kinds of main ingredients were separated from the traditional Chinese medicine (Schisandra) ingredients and 26 peaks were successfully separated from the fermentation broth containing natural lipopeptide antibiotics. The addition of GO material enhanced the interaction between the compound and the monolithic column, increased the binding sites, improved the uniformity of the internal pore structure of the monolithic column, and improved the separation performance of the monolith. Methodologic validation of five ingredients in Schisandra showed that the correlation coefficients of the linear regressions were in the range of 0.9987-0.9997. The intra- and inter-day values of the relative standard deviation for precision were in the range of 0.6-4.1% and 1.1-4.8%, respectively. The values of accuracy (expressed as recovery) were in the range of 97.7-103.2%, 100.5-105.0%, 98.2-101.8%, 101.3-104.1%, and 101.2-103.3% for the 5 ingredients in order. In terms of the relative standard deviation of the retention time, the reproducibility of the monolithic column M1 was <3.7%. The monolithic column based on GO has great potential in chromatographic separation.


Asunto(s)
Lipopéptidos , Metacrilatos , Antibacterianos , Cromatografía Líquida de Alta Presión/métodos , Grafito , Metacrilatos/química , Porosidad , Reproducibilidad de los Resultados
15.
Animal ; 16(12): 100676, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36442324

RESUMEN

Fermented feed has the potential to improve poultry gastrointestinal microecological environment, health condition and production performance. Thus, the present study was undertaken to explore the effects of fermented feed on the laying performance, egg quality, immune function, intestinal morphology and microbiota of laying hens in the late laying cycle. A total of 360 healthy Hy-Line Brown laying hens aged 80 weeks were used to conduct a 56-day study. All hens were randomly separated into two treatment groups, with five replicates of 36 hens each as follows: basal diet containing 0.0% fermented feed (CON) and 20% fermented feed (FF). Subsequent analyses revealed that fermented feed supplementation was associated with significant increases in laying rates together with reduced broken egg rates and feed conversion ratio for hens in FF group (P < 0.05). There were additionally significant increases in both albumen height and Haugh unit values in hens following fermented feed supplementation (P < 0.05). Fermented feed was also associated with increases in duodenal, jejunal and ileac villus height (P < 0.05). Laying hens fed fermented feed had higher immune globulin (Ig)A, IgG, IgM levels (P < 0.01,) and higher interleukin 2, interleukin 6, tumour necrosis factor α and interferon γ (P < 0.05) concentrations than CON. Analysis of the microbiota in these laying hens revealed the alpha diversity was not significantly affected by fermented feed supplementation. Firmicutes abundance was reduced in caecal samples from FF hens relative to those from CON hens (30.61 vs 35.12%, P < 0.05). At the genus level, fermented feed was associated with improvements in relative Lactobacillus, Megasphaera and Peptococcus abundance and decreased Campylobacter abundance in laying hens. These results suggest that fermented feed supplementation may be beneficial to the laying performance, egg quality, immunological function, intestinal villus growth and caecal microecological environment of laying hens at the end of the laying cycle.


Asunto(s)
Suplementos Dietéticos , Microbiota , Animales , Femenino , Alimentación Animal/análisis , Pollos , Dieta/veterinaria , Suplementos Dietéticos/análisis , Inmunidad
16.
Opt Express ; 19(21): 20462-7, 2011 Oct 10.
Artículo en Inglés | MEDLINE | ID: mdl-21997054

RESUMEN

With quasi-periodic microstructures, great enhancement of infrared light absorption of Au film over a broad wavelength band (2.7~15.1 µm) was realized experimentally for the first time. The microstructured Au film was prepared by replica molding of the surface of femtosecond (fs) laser microstructured silicon (black silicon). This unique absorption characteristic is mainly ascribed to good impedance match from free space to Au film. The surface of the sample was examined by X-ray photoelectron spectroscopy (XPS) and the four peaks of absorptance were ascribed to residual polydimethylsiloxane (PDMS), H2SO4, adsorbed water and CO2 in the air, respectively.

17.
Front Nutr ; 8: 706977, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34490325

RESUMEN

The study evaluated the impact of fermentation with Bacillus sp. on the nutritional quality of soybean meal (SBM) and the changes of bacterial community structure during fermentation. High protease-producing strains were screened to degrade SBM macromolecular protein and anti-nutritional factors (ANFs). Unsterilized SBM then underwent an anaerobic solid-state fermentation method to evaluate the effects of fermentation. Results showed that for the nine high-producing protease strains that were screened, acid-soluble protein (ASP) contents in fermented SBM increased, with the highest value found to be 13.48%, which was fermented using strain N-11. N-11 was identified as Bacillus subtilis. N-11 fermentation reduced ANFs such as glycinin and ß-conglycinin by 82.38 and 88.32%, respectively. During N-11 fermentation, the bacterial richness and diversity in SBM increased but not significantly. The high-yield protease strain B. subtilis N-11 selected in this experiment improved the nutritional quality of SBM through fermentation, and it can be used for industrial large-scale production.

18.
Wei Sheng Wu Xue Bao ; 49(4): 492-7, 2009 Apr.
Artículo en Zh | MEDLINE | ID: mdl-19621637

RESUMEN

OBJECTIVE: To purify a single fibrinolytic enzyme from Bacillus pseudomycoides B-60 and to determine its N-terminal sequence and to characterize the fibrinolytic enzyme. METHODS: We examined the fibrinolytic enzyme activity by fibrin plate and purified fibrinolytic enzyme by ammonium sulfate fractional precipitation and DEAE anion exchange chromatography. RESULTS: Obtained a single protein fraction with fibrinolytic activity (BpFE) from B. pseudomycoides B-60. It appeared as a single band in the SDS-PAGE with a relative molecular weight of 34 kDa. The fibrinolytic activity of the protein was stable at 4-50 degrees C and at pH 5-10. The activity sharply decreased above 50 degrees C, and the total loss of activity at pH 3.0. The enzymatic activity was slightly enhanced by the ions of Ca2+, Mg2+, Mn2+, whereas strongly inhibited by Cu2+ ion. Phenylmethyl sulfonyl fluoride (PMSF) could completely inhibit its activity. In addition, the activity improved when the protein was enzymatically hydrolyzed using trypsin and pepsin. The first 15 amino acids of the N-terminal sequence of the enzyme were determined to be VTGTNAVGTGKGVLG. The partial amino acids sequence alignment study of the enzyme from B-60 strain with bacillolysin, neutral protease and hydrolase which were from B. cereus, B. thuringiensis, B. anthracis and Lactobacillus sp. was carried out, and there is a 100% homogeneity between them. CONCLUSION: We obtained a single fibrinolytic enzyme. Through its N-terminal sequence alignment study, a plasmin with high homogeneity to this protein was not found yet. This provided a basis for further study of new thrombolytic drugs.


Asunto(s)
Bacillus/enzimología , Fibrinolíticos/metabolismo , Bacillus/clasificación , Bacillus/genética , Secuencia de Bases , Cromatografía por Intercambio Iónico , ADN Ribosómico/genética , Electroforesis en Gel de Poliacrilamida , Estabilidad de Enzimas , Fibrinolíticos/química , Fibrinolíticos/aislamiento & purificación , Datos de Secuencia Molecular , Homología de Secuencia de Ácido Nucleico
19.
Wei Sheng Wu Xue Bao ; 48(10): 1387-92, 2008 Oct.
Artículo en Zh | MEDLINE | ID: mdl-19160823

RESUMEN

OBJECTIVE: Thrombolytic therapy is a safe and effective treatment for thrombotic diseases. Microorganisms are possible sources of thrombolytic drugs. We purified and characterized fibrinolytic enzyme produced by Bacillus subtilis strain BS-26. METHODS: We examined the fibrinolytic enzyme activity by fibrin plate and purified fibrinolytic enzyme by ammonium sulfate fractional precipitation, anion-exchange chromatography on DEAE-Sepharose Fast Flow and preparative PAGE. RESULTS: The fibrinolytic enzyme of the strain BS-26 was stable blow 50 degrees C and pH5.0-11.0, the optimal temperature was 42 degrees C and optimal pH was 9.0. Ca2+ and Mg2+ ions enhanced the fibrinolytic activity, whereas Cu2+ completely inhibited the enzyme. Phenylmethylsulfonyl fluoride (174.2 microg/mL), chicken ovomucoid (1000 microg/mL) and soybean trypsin inhibitor (1000 microg/mL) could inhibit enzyme activity, which indicated that the enzyme belonged to serine protease group. On plasminogen-free fibrin plates and plasminogen fibrin plates, the fibrinolytic activity had no obvious difference, indicating that the enzyme was a fibrinolytic enzyme which degraded fibrin directly, but not a plasminogen activator which degraded fibrin by activating plasminogen. A fibrinolytic enzyme was purified from the fermentation broth with recovery yield of 3.2%, purification factor of 41.0 fold and the specific activity 8750.0 U/mg. SDS-PAGE analysis of the purified protein showed only one band with molecular mass of 32 kDa. CONCLUSION: A single fibrinolytic enzyme was purified, which provided the basis for large-scale production of fibrinolytic enzyme.


Asunto(s)
Bacillus subtilis/genética , Fibrinolisina/química , Fibrinolisina/aislamiento & purificación , Fibrinolíticos/química , Fibrinolíticos/aislamiento & purificación , Bacillus subtilis/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Estabilidad de Enzimas , Fermentación , Fibrinolisina/genética , Fibrinolíticos/metabolismo , Expresión Génica , Peso Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo
20.
J Phys Condens Matter ; 30(11): 114002, 2018 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-29406312

RESUMEN

Although surface plasmon polaritons (SPPs) have been intensively studied in past years, the transmission/reflection properties of SPPs at an interface between two plasmonic media are still not fully understood. In this article, we employ a mode expansion method (MEM) to systematically study such a problem based on a model system jointing two superlattices, each consisting of a periodic stacking of dielectric and plasmonic slabs with different material properties. Such a generic model can represent two widely used plasmonic structures (i.e. interfaces between two single dielectric/metal systems or between two metal-insulator-metal waveguides) under certain conditions. Our MEM calculations, in excellent agreement with full-wave simulations, uncover the rich physics behind the SPP reflections at generic plasmonic interfaces. In particular, we successfully derive from the MEM several analytical formulas that can quantitatively describe the SPP reflections at different plasmonic interfaces, and show that our formulas exhibit wider applicable regions than previously proposed empirical ones.

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