In this study, we prepared core-shell type nanoparticles of a poly(DL-lactide-co-glycolide) (PLGA) grafted-dextran (DexLG) copolymer with varying graft ratio of PLGA. The synthesis of the DexLG copolymer was confirmed by 1H nuclear magnetic resonance (NMR) spectroscopy. The DexLG copolymer was able to form nanoparticles in water by self-aggregating process, and their particle size was around 50 nm approximately 300 nm according to the graft ratio of PLGA. Morphological observations using a transmission electron microscope (TEM) showed that the nanoparticles of the DexLG copolymer have uniformly spherical shapes. From fluorescence probe study using pyrene as a hydrophobic probe, critical association concentration (CAC) values determined from the fluorescence excitation spectra were increased as increase of DS of PLGA. 1H-NMR spectroscopy using D2O and DMSO approved that DexLG nanoparticles have core-shell structure, i.e. hydrophobic block PLGA consisted inner-core as a drug-incorporating domain and dextran consisted as a hydrated outershell. Drug release rate from DexLG nano-particles became faster in the presence of dextranase in spite of the release rate not being significantly changed at high graft ratio of PLGA. Core-shell type nanoparticles of DexLG copolymer can be used as a colonic drug carrier. In conclusion, size, morphology, and molecular structure of DexLG nanoparticles are available to consider as an oral drug targeting nanoparticles.
A pepper esterase gene (PepEST) that is highly expressed during an incompatible interaction between pepper (Capsicum annuum) and the anthracnose fungus Colletotrichum gloeosporioides has been previously cloned. Glutathione-S-transferase-tagged recombinant PepEST protein expressed in Escherichia coli showed substrate specificity for p-nitrophenyl esters. Inoculation of compatible unripe pepper fruits with C. gloeosporioides spores amended with the recombinant protein did not cause anthracnose symptoms on the fruit. The recombinant protein has no fungicidal activity, but it significantly inhibits appressorium formation of the anthracnose fungus in a dose-dependent manner. An esterase from porcine liver also inhibited appressorium formation, and the recombinant protein inhibited appressorium formation in the rice blast fungus, Magnaporthe grisea. Inhibition of appressorium formation in M. grisea by the recombinant protein was reversible by treatment with cyclic AMP (cAMP) or 1,16-hexadecanediol. The results suggest that the recombinant protein regulates appressorium formation by modulating the cAMP-dependent signaling pathway in this fungus. Taken together, the PepEST esterase activity can inhibit appressorium formation of C. gloeosporioides, which may result in protection of the unripe fruit against the fungus.
The objective of this study was to investigate alginate microencapsulated lactic acid bacteria (LAB) fermentation quality of radish kimchi sample and its potential survivability in different acidic and alkaline environments. Initially, we isolated 45 LAB strains. One of them showed fast growth pattern with potential probiotic and antifungal activities against Aspergillus flavus with a zone of inhibition calculated with 10, 8, 4mm for the 4th, 5th, and 6th day, respectively. Therefore, this strain (KCC-42) was chosen for microencapsulation with alginate biopolymer. It showed potential survivability in in-vitro simulated gastrointestinal fluid and radish kimchi fermentation medium. The survival rate of this free and encapsulated LAB KCC-42 was 6.85 × 105 and 7.48× 105 CFU/ml, respectively; the viability count was significantly higher than nonencapsulated LAB in simulated gastrointestinal juices (acid, bile, and pancreatin) and under radish kimchi fermentation environment. Kimchi sample added with this encapsulated LAB showed increased production of organic acids compared to nonencapsulated LAB sample. Also, the organic acids such as lactic acid, acetic acid, propionic acid, and succinic acid production in fermented kimchi were measured 59mM, 26mM, 14mM, and 0.6mM of g/DW, respectively. The production of metabolites such as lactic acid, acetic acid, and succinic acid and the bacteria population was high in microencapsulated LAB samples compared with free bacteria added kimchi sample. Results of this study indicate that microencapsulated LAB KCC-42 might be a useful strategy to develop products for food and healthcare industries.
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