Optimization of a fed-batch fermentation process for production of bleomycin by Streptomyces mobaraensis ATCC 15003

The objective of this work was to optimize inoculum size and pH for rapid production of bleomycin by Streptomyces mobaraensis ATCC 15003 by fed-batch fermentation. In both batch- and fed-batch fermentations, neither production period for bleomycin by S. mobaraensis nor the amount of BLM were affected by increasing the inoculum size from regular 10 to 30% (v/v) level. A fed-batch bioreactor not only shortened the lag phase of BLM production from 114 to 60 h, but also fed-batch fermentationenhanced BLM production when combined with an appropriate pH profile with no effect on the amount produced. Due to the substrate inhibition that takes place at high levels of carbon source, fed-batch fermentation was proposed as a better alternative for BLM production. The combined effects of batch and fed-batch fermentation and various pH profiles on BLM production in a bioreactor were evaluated. The tested pH profiles included; (1) a constant pH profile at 6.8 (profile1); (2) a constant pH profile witha period of auto-acidification for 72 h (profile2) and (3) a step-wise pH profile with pH adjustment every 24 h (profile3). When profile 3 was applied, fed-batch fermentation enhanced BLM production in the bioreactor and yielded about two-fold higher BLM concentration than the irrespective batch fermentation. On the other hand, constant pH profile or that which included one period of autoacidification (profiles 1 and 2) resulted in an insignificantly difference BLM production in fed-batch fermentation. Overall, this study suggested that fed-batch fermentation can be successfully used to enhance BLM production in bioreactor especially with fluctuated pH-profile.Key words: Bleomycin, Streptomyces mobaraensis, rapid production, fed-batch fermentation

Improvement and enhancement of clavulanic acid production in Streptomyces clavuligerus using vegetable oils

Clavulanic acid (CA) is a potent inhibitor of β-lactamases. Oil can be used as a source of carbon and energy in CA production. To develop a policy for enhanced production of CA, the selection of a production medium and an optimum pH, different vegetable oils as carbon source have been used. The effects of different medium containing vegetable oil on cell growth and CA yield production during the fermentation of Streptomyces clavuligerus ATCC 27064 were demonstrated. In this study, three out of eight tested oils supported CA production. Medium containing olive oil showed two-fold higher CA yield than glycerol containing medium. The highest productivity was obtained at initial pH 7. We concluded that using olive oil as a sole source of carbon and energy for cultivation of S. clavuligerus is a promising strategy for CA production. It has several scientific advantages and economic benefits that lead to increased antibiotic titre and can be considered as a cheaper alternative compared to carbohydrates. The results of this study can be applied for the efficient production of β-lactamase inhibitory antibiotics.Key words: Clavulanic acid, enzymes, β-lactamases, vegetable oils, Streptomyces clavuligerus

Formulation and Optimization of Zidovudine Niosomes

Zidovudine (AZT) is commonly used to treat patients with AIDS, but it is limited by toxicity and high dosing needs. Alternative formulations have been proposed to overcome these drawbacks. The objective of this study was to evaluate process-related variables like hydration and sonication time, rotation speed of evaporation flask, and the effects of charge-inducing agent and centrifugation on zidovudine entrapment and release from niosomes. Formulation of zidovudine niosomes was optimized by altering the proportions of Tween, Span and cholesterol. The effect of process–related variables like hydration time, sonication time, charge-inducing agent, centrifugation and rotational speed of evaporation flask on zidovudine entrapment and release from niosomes was evaluated. The effect of changes in osmotic shock and viscosity were also evaluated. Non-sonicated niosomes were in the size range of 2-3.5 μm and sonicated niosomes formulated with Tween 80 and dicetylphosphate (DCP) had a mean diameter of 801 nm. Zidovudine niosomes formulated with Tween 80 entrapped high amounts of drug and the addition of DCP enhanced drug release for a longer time (88.72% over 12 h). The mechanism of release from Tween 80 formulation was the Fickian type and obeyed first-order release kinetics. Niosomes can be formulated by proper adjustment of process parameters to enhance zidovudine entrapment and sustainability of release. These improvements in zidovudine formulation may be useful in developing a more effective AIDS therapy

Evaluation of alternative strategies to optimize ketorolac transdermal delivery

In the present study, 2 alternative strategies to optimize ketorolac transdermal delivery, namely, prodrugs (polyoxyethylene glycol ester derivatives, I–IV) and nanostructured lipid carriers (NLC) were investigated. The synthesized prodrugs were chemically stable and easily degraded to the parent drug in human plasma. Ketorolac-loaded NLC with high drug content could be successfully prepared. The obtained products formulated into gels showed a different trend of drug permeation through human stratum corneum and epidermis. Particularly, skin permeation of ester prodrugs was significantly enhanced, apart from ester IV, compared with ketorolac, while the results of drug release from NLC outlined that these carriers were ineffective in increasing ketorolac percutaneous absorption owing to a higher degree of mutual interaction between the drug and carrier lipid matrix. Polyoxyethylene glycol esterification confirmed to be a suitable approach to enhance ketorolac transdermal delivery, while NLC seemed more appropriate for sustained release owing to the possible formation of a drug reservoir into the skin