Recombinant human growth hormone (rhGH) was encapsulated within poly(D,L-lactic-co-glycolic acid) microspheres by a double emulsion solvent evaporation method. A mixture of methylene chloride and ethyl acetate in varying volume ratios was used for the microsphere preparation. Protein release profiles from three different microsphere formulations demonstrated initial burst effects ranging from 28.2% to 54.7% after a 1-day incubation and exhibited no further significant releases up to 19 days. This was because the encapsulated rhGH with the microspheres was largely aggregated in a noncovalent fashion during the formulation. Nonaggregated water soluble rhGH species within the microspheres are likely to be responsible for the rapid release upon incubation. The initially released rhGH in the incubation medium, however, was composed of mostly monomer species with a small amount of dimer as probed by size-exclusion chromatography. Circular dichroism spectra of the initially released rhGH in the medium revealed that the conformation of the released rhGH was correctly folded relative to that of native rhGH, with little variation in alpha-helix contents depending on the formulations. The "nonrelease" mechanism after the initial burst release was attributed to nonspontaneously dissociable noncovalent protein aggregation and surface adsorption of rhGH present within the microspheres.

In order to develop environment friendly fungicide for the control of citrus green mold (Penicillium digitatum) using endophytic bacteria, the 21 bacterial isolates were isolated from citrus leaves in seven different orchards in Jeju Province. Among the 21 bacterial isolates, 5 bacterial isolates presented antifungal activity against green mold pathogen P. digitatum. The CB3 isolate, which showed the most strong antagonistic effect, was selected through opposite culture against the pathogen. The rod-shaped, gram-positive bacterium CB3 was identified as Bacillus velezensis based on morphological, physiological characteristics, 16S rDNA, and gyr A gene sequence analysis. The isolate CB3 showed strong antifungal activity against two citrus postharvest pathogen P. digitatum. Citrus fruits were treated by wound inoculation with P. digitatum pathogen, and the control efficacy of CB3 culture broth was 66.7% (1 × 10 8 cfu/ml). In conclusion, The stability of CB3 and its strong antifungal activity also lead us to believe that it has potential for application as an environment friendly biological control agent.

Slightly acidic hypochlorous water (SAHW) is well known for having a powerful and broad spectrum antimicrobial activity, and is harmless to the environment and humans. SAHW (pH 5~6.5, 20~30 ppm available chlorine concentration) was generated by electrolysis of dilute solution of HCl (4%) in a chamber of a non-membrane electrolytic cell. Our objective was to determine SAHW has a potential fungicidal activity on some phytopathogenic fungi. Spores of Botrytis cinerea, Colletotrichum acutatum and Phytophthora capsici were not culturable on agar media at approximately 10 seconds after treatment by SAHW. However, inactivation of Penicillium hirsutum was required over 3 min. Dilution of SAHW with sterilized distilled water (SDW) at the ratio of 1:1 (SAHW:SDW) against C. acutatum showed 100% inactivation but, the efficacy in 1:2 decreased until 63.2%. Control value of SAHW was 70.4% against C. acutatum on pepper fruits when applied upto 24 h postinoculation. SAHW has a powerful and wide spectrum antifungal activity and could be applied as a potential alternative to fungicidal agent for control of plant disease.

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