Background: Amylases are among the most important enzymes which are of great significance for biotechnology and have almost completely replaced chemical hydrolysis of starch in the starch processing industry. The present study was concerned with the production and optimization of extracellular α-amylase from Bacillus sp. NRC22017. Results: The effect of various fermentation conditions on α-amylase production through shake-flask culture was investigated. Bacterial strain produces α-amylase was isolated from water in Wadi El-Natron. Based on microbiological, biochemical tests, and 16S rRNA gene sequences, the isolate was identified as Bacillus sp. NRC22017 and was later used for further studies. Maximum yield of α-amylase is 15.15 ± 0.47 U/ml from Bacillus sp. NRC22017; this strain is characterized with high temperature and high salinity in cultivated culture, and achieved maximum yield of α-amylase at pH 6.0 with inoculum size of 500 μl at 45°C and aerobically incubation period of 72 h. The optimum volume of the fermentation medium was found to be 20 ml in 100 ml Erlenmeyer flask; the best starch and meat extract plus peptone concentration that provided the highest enzyme production from Bacillus sp. NRC22017 were found to be 2% and 1.05% (w/v) respectively. Conclusion: Enzyme production was higher after optimizing the production conditions as compared to the basal medium.
Trehalose synthase (TSII) from Corynebacterium nitrilophilus NRC was successively purified by ammonium sulphate precipitation, ion exchange chromatography on DEAE-cellulose and gel filtration chromatography on Sephadex G-100 columns. The specific activity of the trehalose synthase was increased *200-fold, from 0.14 U mg -1 protein to 28.3 U mg -1 protein. TSII was found to be a monomeric protein with a molecular weight of 67-69 kDa. Characterization of the enzyme exhibited optimum pH and temperature were 7.5 and 35°C, respectively. The purified enzyme was stable from pH 6.6 to 7.8 and able to prolong its thermal stability up to 35°C. The enzyme activity was inhibited strongly by Zn 2? , Hg 2? and Cu 2? and moderately by Ba 2? , Fe 2? , Pb 2? and Ni 2? . Other metal ions Ca 2? , Mg 2? , Co 2? , Mn 2? and EDTA had almost no effect.
Pertussis specific antibodies were studied with respect to quality and quantity in a cohort of apparently healthy Egyptian children and adolescents, with their age range between 1 and 18 years, in an attempt to get a close and clear insight into the current humoral immunization status in this specified group and to try find a relation between the antibody levels and their avidities in eradication of this devastating infectious disease. Our results showed that avidity increase was most marked in young school children (6–8 years) where it seemed to reach a plateau in older children and adolescents. Antibody titer was highest in toddlers (1–2 years) and young school children (6–8 years) groups, most probably following vaccination and/or booster doses. Among children aged 1–5 years, 28% had highly avid and 50% had high titer antibodies, whereas in adolescents aged 13–18 years, 70% had highly avid antibodies and only 30% had high titer antibodies. The results clearly demonstrated that while levels of anti-Bordetella pertussis (B. pertussis) antibodies wane with growing age, the avidity seems to increase, to a plateau, irrespective of further antigen exposure in a pattern showing complete independence of avidity on concentration. The present study draws attention to the importance of avidity measurements, together with conventional ELISAs, for evaluating immunity against pertussis. Being based on a limited sample size, it could open doors for larger-scale surveys to be possible indicators for the need and timing of booster vaccination doses among Egyptians.
Microbial exopolysaccharides (EPSs) extracted from lactic acid bacteria (LAB) are generally recognized as safe. They have earned popularity in recent years because of their exceptional biological features. Therefore, the present study main focus was to study EPS-production from probiotic LAB and to investigate their antioxidant and burn wound healing efficacy. Seventeen LAB were isolated from different food samples. All of them showed EPS-producing abilities ranging from 1.75 ± 0.05 to 4.32 ± 0.12 g/l. RO30 isolate (from Romi cheese) was chosen, due to its ability to produce the highest EPS yield (4.23 ± 0.12 g/l). The 16S rDNA sequencing showed it belonged to the Lactiplantibacillus plantarum group and was further identified as L. plantarum RO30 with accession number OL757866. It displayed well in vitro probiotic properties. REPS was extracted and characterized. The existence of COO−, OH and amide groups corresponding to typical EPSs was confirmed via FTIR. It was constituted of glucuronic acid, mannose, glucose, and arabinose in a molar ratio of 2.2:0.1:0.5:0.1, respectively. The average molecular weight was 4.96 × 104 g/mol. In vitro antioxidant assays showed that the REPS possesses a DPPH radical scavenging ability of 43.60% at 5 mg/ml, reducing power of 1.108 at 10 mg/ml, and iron chelation activity of 72.49% and 89.78% at 5 mg/ml and 10 mg/ml, respectively. The healing efficacy of REPS on burn wound models in albino Wistar rats showed that REPS at 0.5% (w/w) concentration stimulated the process of healing in burn areas. The results suggested that REPS might be useful as a burn wound healing agent.
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