Purification and characterization of a milk-clotting protease from Mucor pusillus: Method comparison

Crude enzymatic extract obtained from five fermentations (300 g of wheat bran) was characterized by a clotting activity of 0.34 ± 0.08 UP/ml with a strength ratio of 1/1: 200. The comparative study of the summaries from 2 purification protocols showed that it is possible to recover 6% of the initial proteins with a 44.54% activity after gel filtration (protocol I), which appeared more technically sound when compared to ion-exchange (1.80% of total proteins with a 23% performance) (protocol II). The proteinhomogeneity (a single electrophoretic band) of the monomeric protease was confirmed by both methods after precipitation with 80% saturated ammonium sulphate. Moreover, the fractional precipitation technique with this salt (40 and 80%) was useless in the experimental conditionsemployed and an important loss of activity was observed (28.53%) with a 3-fold purification. In another part of the study, without ammonium sulphate precipitation, the gel filtration enabled the elimination of almost 97% of the inactive proteins and improved the activity performance by 55.13%, while multiplying the specific activity of the coagulant by a factor of 20.88 against a 6.75-fold purification with ionexchange and the appearance of a more or less 20 kDa peptide after electrophoresis. The proteolytic activity of the purified extracts had a similar appearance to a more pronounced kinetic when compared with the reference rennet. The purification protocols did not seem to have an impact on the isolatedprotease activity.Key words: Mucor pusillus, protease, purification, enzymatic performance, electrophoresis, milk clotting,rennet

The immune system and the impact of zinc during aging

The trace element zinc is essential for the immune system, and zinc deficiency affects multiple aspects of innate and adaptive immunity. There are remarkable parallels in the immunological changes during aging and zinc deficiency, including a reduction in the activity of the thymus and thymic hormones, a shift of the T helper cell balance toward T helper type 2 cells, decreased response to vaccination, and impaired functions of innate immune cells. Many studies confirm a decline of zinc levels with age. Most of these studies do not classify the majority of elderly as zinc deficient, but even marginal zinc deprivation can affect immune function. Consequently, oral zinc supplementation demonstrates the potential to improve immunity and efficiently downregulates chronic inflammatory responses in the elderly. These data indicate that a wide prevalence of marginal zinc deficiency in elderly people may contribute to immunosenescence

Purification and characterization of a milk-clotting protease from Mucor pusillus : method comparison

Crude enzymatic extract obtained from five fermentations (300 g of wheat bran) was characterized by a clotting activity of 0.34 ± 0.08 UP/ml with a strength ratio of 1/1: 200. The comparative study of the summaries from 2 purification protocols showed that it is possible to recover 6% of the initial proteins with a 44.54% activity after gel filtration (protocol I), which appeared more technically sound when compared to ion-exchange (1.80% of total proteins with a 23% performance) (protocol II). The protein homogeneity (a single electrophoretic band) of the monomeric protease was confirmed by both methods after precipitation with 80% saturated ammonium sulphate. Moreover, the fractional precipitation technique with this salt (40 and 80%) was useless in the experimental conditions employed and an important loss of activity was observed (28.53%) with a 3-fold purification. In another part of the study, without ammonium sulphate precipitation, the gel filtration enabled the elimination of almost 97% of the inactive proteins and improved the activity performance by 55.13%, while multiplying the specific activity of the coagulant by a factor of 20.88 against a 6.75-fold purification with ionexchange and the appearance of a more or less 20 kDa peptide after electrophoresis. The proteolytic activity of the purified extracts had a similar appearance to a more pronounced kinetic when compared with the reference rennet. The purification protocols did not seem to have an impact on the isolated protease activit

Extracellular protease from mucor pusillus : purfication and characterization

Extracellular protease from Mucor pusillus was purified 18-fold with 7.56% recovery by ion-exchange chromatography and gel filtration. The enzyme was found to be monomeric in nature, having a molecular mass of 49 kDa. The enzyme acted optimally at 50°C and was stable in the temperature range 30–50°C. It was completely inactivated by heating for 30 min at 65°C. The optimum of activity for the purified extract was observed at milk CaCl2 concentration of 0.02 m and at milk pH of 5. These properties, except for temperature, were similar to those of renne

Does BMI Really Alter the Hormonal Profile in Infertile Women? Retrospective Study in the Region of Sidi Bel Abbes (West Algeria)

Background: In last few years .; it’s  increasingly being recognized  that  Reproductive function is controlled by the hypothalamic–pituitary– gonadal axis, which is regulated by numerous endogenous and environmental factors  such us adipose accumulation in obesity  contributing to reproductive failure  such as menstrual disorders and infertility, gestational failure and obstetric complications, and infertility , Distinct changes in circulating sex hormones appear to underline these abnormalities. The objective: The aim of this study was to elucidate the possible correlation between body mass index as fatness indicator and hormonal profile in infertile women from the west of Algeria. To identify the impact of overweight  and obesity on female hormonal profile ; we  conducted a prospective study  measuring pituitary hormones  (FSH  and LH and prolactin ) , steroid sex hormones ( progesterone , testosterone and estradiol ) , anti miulleian hormone , and thyroidal hormones (FT3 , FT4 and TSH) in 360 women  consulting for subfertility in private genecologycal and obstetrical centers in SID BELABESS (West of Algeria ) Result: Our study showed that the majority of patients were aged between 20 and 29 years, representing a percentage of 47.8%, with the average age in sample (31.65 ± 6,93ans). The majority of subjects was obese   46.4%, or overweight (39.4%) with an average BMI of (29.76 ± 4,85Kg / m2). No statistically significant association was found between the BMI as obesity indicator with hormonal levels of pituitary hormones  (FSH  and LH and prolactin ) , steroid sex hormones ( progesterone , testosterone and estradiol ) , anti miulleian hormone , and thyroidal hormones (FT3 , FT4 and TSH). A negative and statistically significant correlation between the age of the patients and the AMH level (R = -0.60, P <0.01) was noted. Conclusion: The results of our study showed that the majority of women included in our study had a high BMI but no statistical significant difference was found between underweight , normal , overweight and obese women , that’s why more studies should be conducted to elucidate the in which level does obesity impair the reproductive outcomes. Keywords: Female infertility, hormonal profile, BMI, SBA, Algeria