The chemical composition of the essential oils obtained from stem/leaves and flowers/fruits of Ferula szovitsiana (Umbelliferae) were analysed by GC and GC-MS. The composition of the oils samples from stem/leaves and flower/fruits were compared: 51 and 47 compounds were identified in the essential oils of stem/leaves and flowers/ fruits, representing 97.7% and 95.9% of the total oils, respectively. Both essential oils were rich in monoterpenes (66.7% and 62.1%, respectively) with neryl acetate as major component (33.0% and 41.5%, respectively). Other abundant compounds for stem/leaves oil were β β β β β-caryophyllene (8.9%), α α α α α-pinene (8.0%), β β β β β -pinene (6.7%), bicyclogermacrene (4.5%), caryophyllene oxide (4.1%), limonene (4.6%) and α α α α α-terpineol (3.2%). These values for flowers/fruits oil were bicyclogermacrene (9.0%), α α α α α-pinene (5.5%), β β β β β -pinene (3.9%), γ γ γ γ γ -cadinene (3.5%), and calarene (3.2%). Furthermore, the oils were tested against Gram-positive, Gram-negative bacteria and two pathogenic fungi. It was found that oil from F. szovitsiana represents the most potent antimicrobial candidates with MIC of 1.25 mg/disk for Bacillus subtilis. Copyright
The methylotrophic yeast Pichia pastoris is a well-established expression host, which is often used in the production of protein pharmaceuticals. This work aimed to evaluate the effect of various concentrations of ascorbic acid in mixed feeding strategy with sorbitol/methanol on productivity of recombinant human growth hormone (r-hGH). The relevant concentration of ascorbic acid (5, 10, or 20 mmol) and 50 g/L sorbitol were added in batch-wise mode to the medium at the beginning of induction phase. The rate of methanol addition was increased stepwise during the first 12 h of production and then kept constant. Total protein and r-hGH concentrations were analyzed and the results compared with sorbitol/methanol feeding using one-way analysis of variance. Moreover, an effective clarification process using activated carbon was developed to remove process contaminants like pigments and endotoxins. Finally, a three-step chromatographic process was applied to purify the product. According to the obtained results, addition of 10 mmol ascorbic acid to sorbitol/methanol co-feeding could significantly increase cell biomass (1.7 fold), total protein (1.14 fold), and r-hGH concentration (1.43 fold). One percent activated carbon could significantly decrease pigments and endotoxins without any significant changes in r-hGH assay. The result of the study concluded that ascorbic acid in combination with sorbitol could effectively enhance the productivity of r-hGH. This study also demonstrated that activated carbon clarification is a simple method for efficient removal of endotoxin and pigment in production of recombinant protein in the yeast expression system.
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