Light calcium carbonate improves pullulan biosynthesis by Aureobasidium pullulans under high concentration of sugar.

The effects of light calcium carbonate (CaCO3) on pullulan biosynthesis by Aureobasidium pullulans NCPS2016 were investigated. Light CaCO3 enhanced pullulan production by 12.4 % when added to the low concentration of fructose broth compared with K2HPO4. Pullulan production was further improved when increasing both the concentrations of light CaCO3 and fructose. Compared to K2HPO4, light CaCO3 improved the activities of UDP-glucose pyrophosphorylase, α-phosphoglucose mutase, UDP-glucosyltransferase, and glucosyltransferase relevant to pullulan biosynthesis, and the gene transcriptional levels of UDP-glucose pyrophosphorylase, α-phosphoglucose mutase, UDP-glucosyltransferase, and glucose kinase were enhanced. During 30-liter fermentation, 144.3 g/L of purified pullulan was produced from 200 g/L of fructose and 15 g/L of light CaCO3 within 168 h, with the yield and productivity of 0.72 g/g and 0.86 g/L/h respectively. This is the first report that light CaCO3 improves pullulan production significantly.

High-level production of pullulan from high concentration of glucose by mutagenesis and adaptive laboratory evolution of Aureobasidium pullulans.

The cost of carbon sources and the low efficiency of the fermentation titer limit the industrial application of pullulan. In this study, a hypertonic-tolerant strain with efficient utilization of glucose was obtained using a double strategy. Initially, a strain for efficient synthesis of pullulan from glucose was generated by mutagenesis. Subsequently, the mutant was directionally evolved on the plate containing a high glucose concentration to enhance high osmotic resistance. The enzyme activities and the transcriptional levels involved in pullulan biosynthesis and high osmotic tolerance in mutants were increased. Nitrogen source and inorganic salts also significantly affected the production of pullulan by M233-20 from high concentration of glucose. The pullulan titer of 162.1 g/L was obtained using the response surface methodology in the flask. The strain M233-20 produced 162.3 g/L pullulan in a 30-L bioreactor with a yield of 0.82 g/g glucose. Hence, this work provides a potential industrial pullulan producer M233-20 and a strategy to develop excellent strain.

Identification of Lipopeptide Iturin A Produced by Bacillus amyloliquefaciens NCPSJ7 and Its Antifungal Activities against Fusarium oxysporum f. sp. niveum.

Bacillus amyloliquefaciens NCPSJ7 showed potential fungicidal activities for the effective control of fungal infection. From the PCR test, the key genes (srfAA, sfp, fenD, bmyB, ituD, and ituC) were detected in B. amyloliquefaciens NCPSJ7. These genes were closely related to the lipopeptides (LPs) synthesis. Next, three LPs families were identified with liquid chromatography-mass spectrometry (LC/MS), including iturin A, fengycin A, and surfactin. After purification with C18, the main active antifungal compound was proven to be C14-iturin A by ESI-HRMS, which has significant activities against fungi. These results proved that C14-iturin A played an important role in inhibiting the growth of fungi for B. amyloliquefaciens NCPSJ7. Furthermore, the isolated LP could inhibit mycelial growth and conidia germination at 30 µg/mL. SEM allowed us to observe that mycelial morphology and conidia germination were also affected. The mycelial ultrastructure TEM observations showed that the external electron-dense outer layer cell wall, which mainly consisted of glycoproteins, was affected. Furthermore, swollen mitochondria, enriched glycogen, and increased vacuoles were also found. LP also affected the intact wall and membranes, leading to their increased permeability, which was proved by propidium iodide (PI) staining and conductivity measurements. Meanwhile, the ergosterol, which has an affinity for iturin A, also increased. These results indicated that LP caused fungal dysfunction and membrane permeability increase, leading to fungal inhibition. Identifying and studying LPs is important in exploring the fungicidal activities of B. amyloliquefaciens, which promotes the use of B. amyloliquefaciens NCPSJ7 as a potential candidate for biocontrol.

Divergent Conversion of Double Isocyanides with Alkenyl Bromide to Polysubstituted Pyrroles and 4-Imino-4,5-dihydropyrrolo[3,4-b]pyrrol-6(1H)-one Derivatives by Pd-Catalyzed Tandem Cyclization Reactions.

Herein a novel and concise approach to pyrrole skeletons via Pd-catalyzed tandem cyclization reactions is investigated. The substrates for the transformation could be readily prepared by phosphoric acid-catalyzed Ugi reactions with available starting materials. In this strategy, two isocyanides participate in sequential isocyanide insertion reactions, and the chemoselectivity of the products is regulated by the steric hindrance of the isocyanide. A plausible mechanism for the formation of the corresponding adducts is proposed.

Eco-Friendly and Facile Synthesis of Antioxidant, Antibacterial and Anticancer Dihydromyricetin-Mediated Silver Nanoparticles.

BACKGROUND: Dihydromyricetin (DMY), a natural flavonoid, has reportedly antibacterial, antioxidant, anticancer and other properties. In the present study, DMY was used as a reducing agent and stabilizer to synthesize silver nanoparticles (AgNPs), and the optimal conditions for its synthesis were studied. The DMY-AgNPs were investigated for their DPPH scavenging properties and their potential against human pathogenic and food-borne bacteria viz. Escherichia coli (E. coli), and Salmonella. In addition, DMY-AgNPs also showed excellent inhibitory effects on cancer Hela, HepG2 and MDA-MB-231 cell lines. METHODS: The dihydromyricetin-mediated AgNPs (DMY-AgNPs) were characterized by ultraviolet-visible spectrophotometer (UV-Vis spectra), scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and X-ray powder diffraction (XRD). Antioxidant activity of DMY-AgNPs was determined by 1.1-diphenyl-2-picrylhydrazyl (DPPH) scavenging. The antibacterial activity was determined by 96-well plate (AGAR) gradient dilution, while anticancer potential was determined by MTT assay. RESULTS: The results showed that the dispersion of AgNPs had the maximum UV-visible absorption at about 410 nm. The synthesized nanoparticles were almost spherical. FTIR was used to identify functional groups that may lead to the transformation of metal ions into nanoparticles. The results showed that the prepared AgNPs were coated with biological molecules in the extraction solution. The biosynthesized DMY-AgNPs exhibited good antioxidant properties, at various concentrations (0.01-0.1mg/mL), the free radical scavenging rate was about 56-92%. Furthermore, DMY-AgNPs possessed good antibacterial properties against Escherichia coli (E. coli), and Salmonella at room temperature. The minimum inhibitory concentrations (MIC) were 10-6 g/L, and 10-4 g/L, respectively. The bioactivity of DMY-mediated AgNPs was studied using MTT assay against Hela, HepG2 and MDA-MB-231 cancer cell lines, and all showed good inhibitory effects. CONCLUSION: The present study provides a green approach for the synthesis of DMY-AgNPs which exhibited stronger antioxidant, antibacterial and anticancer properties compared to the dihydromyricetin. DMY-AgNPs can serve as an economical, efficient, and effective antimicrobial material for its applications in food and pharmaceutical fields.

Green and Facile Synthesis and Antioxidant and Antibacterial Evaluation of Dietary Myricetin-Mediated Silver Nanoparticles.

Myricetin (MY) is a dietary flavonoid which exhibits a wide spectrum of biological properties, viz., antibacterial, antioxidant, anticancer, and so forth. The lower solubility in aqueous medium and hence lesser bioavailability of MY limits the use of such dietary flavonoids in further in vivo research. To overcome bioavailability limitations, a number of drug-delivery systems are being investigated. Herein, MY-mediated silver nanoparticles (MY-AgNPs) were synthesized by a green approach to improve the therapeutic efficacy of MY. MY-AgNPs were characterized by ultraviolet-visible spectroscopy (UV-Vis), scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectroscopy, and X-ray powder diffraction (XRD). The results showed that the dispersion of AgNPs had the maximum UV-vis absorption at about 410 nm. The synthesized nanoparticles were almost spherical. MY-AgNPs were further investigated against human pathogenic bacteria, and their antioxidant potential was also determined. The free radical scavenging rate was about 60-87%. MY-AgNPs had good antibacterial activity against Escherichia coli and Salmonella at room temperature with minimum inhibitory concentrations of 10-4 and 10-5 g/L, respectively.

Application of antagonist Bacillus amyloliquefaciens NCPSJ7 against Botrytis cinerea in postharvest Red Globe grapes.

We investigated the effects and possible mechanisms of Bacillus amyloliquefaciens NCPSJ7 against the gray mold caused by Botrytis cinerea in the postharvest Red Globe grapes. The disease incidence, lesion diameter, decay index, and some resistance-related enzymes were evaluated. The antioxidant capacity of grape treated with 1 × 104 CFU/ml B. cinerea alone and combined with 1 × 107 CFU/ml NCPSJ7 was also determined. The results showed that NCPSJ7 + B. cinerea reduced the disease incidence, lesion diameter, and decay index of postharvest grapes and enhanced the activities of polyphenol oxidase, peroxidase, chitinase, and ß-1,3-glucanase during different storage periods. Furthermore, the oxidative resistance, demonstrated by an escalating trend in the total phenolic content, DPPH free radical clearance rate, reducing power, and superoxide anion clearance rate after lesion presence, was improved. However, NCPSJ7 showed an inhibitory effect on gray mold, but resulted in the reduced antioxidant capacity in the grapes.

Fast atrazine degradation by the mixed cultures enriched from activated sludge and analysis of their microbial community succession.

In this study, fast atrazine degradation by the mixed bacterial cultures from sewage sludge was investigated. The acquired activated cultures showed great capability in atrazine degradation. The biodegradation process was well fitted into a pseudo-first reaction kinetic model. Atrazine could inhibit the propagation of ammonium oxidation bacteria and nitrite oxidation bacteria, decreasing the ammonium removal rate and the accumulation of nitrite. Only 162-172 reads of Nitrosomonadaceae and no Nitrospirales were detected after atrazine was exposed to the mixed cultures. The bacterial community structures in the cultures under different inoculation conditions (with or without atrazine) were investigated to explore the mechanism of atrazine degradation. Our results show that the genera Thiobacillus and Ferruginibacter were the most possible candidates responsible for the degradation of atrazine.

Screening and Identifying Antioxidative Components in Ginkgo biloba Pollen by DPPH-HPLC-PAD Coupled with HPLC-ESI-MS2.

The Ginkgo biloba is one of ancient trees that exists from billions of years ago, its leaf and nut are used as herbs and foods in China, while so far its pollen does not have any application except pollination. In order to evaluate the antioxidant activity of Ginkgo biloba pollen, and rapidly screen its antioxidative components, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability, total flavonoid, total phenol, and proanthocyanidin of Ginkgo biloba pollen were determined and compared with those of Ginkgo biloba leaf and nut, and the off-line DPPH-HPLC-PAD and HPLC-ESI-MS2 were applied for screening and identifying the antioxidant flavonoids in Ginkgo biloba pollen. The results showed that the DPPH scavenging ability of Ginkgo biloba pollen was much higher than Ginkgo biloba nut, but lower than Ginkgo biloba leaf, while the total content of flavonoid in Ginkgo biloba pollen was approximately 4.37 times higher than in Ginkgo biloba leaf. Further studies found that the major flavonol aglycone in Ginkgo biloba pollen was kaempferol, which accounted for 96.71% of the total aglycones (includes quercetin, kaempferol and isorhamnetin), and the main flavonoid components in Ginkgo biloba pollen were flavonoid glycosides. Finally, ten antioxidant peaks were screened and identified to be flavonoids (including kaempferol and nine flavonoid glycosides), so flavonoids were likely to be the main antioxidant components in GP, and among them, three novel kaempferol glycosides (peaks 1, 2, and 3) were found in Ginkgo biloba pollen for the first time, which had never been found in Ginkgo biloba.

Screening natural antioxidants in peanut shell using DPPH-HPLC-DAD-TOF/MS methods.

Peanut shell, a byproduct in oil production, is rich in natural antioxidants. Here, a rapid and efficient method using DPPH-HPLC-DAD-TOF/MS was used for the first time to screen antioxidants in peanut shell. The method is based on the hypothesis that upon reaction with 1, 1-diphenyl-2-picrylhydrazyl (DPPH), the peak areas of compounds with potential antioxidant activities in the HPLC chromatogram will be significantly reduced or disappeared, and the identity confirmation could be achieved by HPLC-DAD-TOF/MS technique. With this method, three compounds possessing potential antioxidant activities were found abundantly in the methanolic extract of peanut shell. They were identified as 5,7-dihydroxychromone, eriodictyol, and luteolin. The contents of these compounds were 0.59, 0.92, and 2.36 mg/g, respectively, and luteolin possessed the strongest radical scavenging capacity. DPPH-HPLC-DAD-TOF/MS assay facilitated rapid identification and determination of natural antioxidants in peanut shell, which may be helpful for value-added utilization of peanut processing byproducts.