Cloning, expression, purification and CD analysis of recombinant human betatrophin

Betatrophin is a member of the angiopoietin-like (ANGPTL) family that has been implicated in both triglyceride and glucose metabolism. The physiological functions and molecular targets of this protein remain largely unknown; hence, a purified available protein would aid study of the exact role of betatrophin in lipid or glucose metabolism. In this study, we cloned the full-length cDNA of betatrophin from a human liver cDNA library. Betatrophin was expressed in the pET-21b-E. coli Bl21 (DE3) system and purified by immobilized metal-affinity chromatography and ion-exchange chromatography. Circular dichroism spectroscopy revealed α-helix as the major regular secondary structure in recombinant betatrophin. The production method is based on commonly available resources; therefore, it can be readily implemented

Cloning, expression, purification and CD analysis of recombinant human betatrophin

Betatrophin is a member of the angiopoietin-like (ANGPTL) family that has been implicated in both triglyceride and glucose metabolism. The physiological functions and molecular targets of this protein remain largely unknown; hence, a purified available protein would aid study of the exact role of betatrophin in lipid or glucose metabolism. In this study, we cloned the full-length cDNA of betatrophin from a human liver cDNA library. Betatrophin was expressed in the pET-21b-E. coli Bl21 (DE3) system and purified by immobilized metal-affinity chromatography and ion-exchange chromatography. Circular dichroism spectroscopy revealed α-helix as the major regular secondary structure in recombinant betatrophin. The production method is based on commonly available resources; therefore, it can be readily implemented

Inhibitory effect of ferula persica, ginkgo biloba, nelumbo nucifera, and dicyclomine on the activity of pancreatic lipase

Background and purpose: Pancreatic lipase is a major digestive enzyme in digestion and absorption of fats. Inhibition of the pancreatic lipase activity has always been one of the goals of researchers to control obesity and related disorders. Current inhibitory agents have several side effects. The aim of this study was to investigate the inhibitory effect of Ferula persica, Ginkgo biloba, Nelumbo nucifera, and Dicyclomine on pancreatic lipase activity. Materials and methods: In this experimental study, Ferula persica, Ginkgo biloba, and Nelumbo nucifera were extracted by soxhlet method. Methanolic or aqueous extracts of plants and dicyclomine at 10, 25, 50, 100, 200, and 400 μg/ml were prepared and their inhibitory effect on a fixed concentration of commercial pancreatic lipase were investigated. The combined extracts of Ferula persica and Nelumbo nucifera were also applied to the enzyme at the ratio of 1:3, 2:2, 3:1, and without quotas. Lipase activity was measured based on the release of methyl resorphine as colorimetric assay. Results: Extracts of Ferula persica and Nelumbo nucifera alone or combined in a ratio of 1:2 at 50 μg/ml led to further decrease in pancreatic lipase activity compared to the unexposed enzyme. Ginkgo biloba and Dicyclomine did not show any considerable inhibitory effect at concentrations and ratios studied. Conclusion: The extracts of Ferula persica and Nelumbo nucifera, alone or in combination, showed inhibitory effect on pancreatic lipase activity. Further studies, especially clinical trials are suggested to evaluate the efficacy and safety of these compounds

Calprotectin Pegylation Enhanced Its Physical and Structural Properties

Calprotectin is member of the S-100 protein family with a wide plethora of intra-and extracellular functions. Anticancer activities, antimicrobial effects and being a qualified disease marker are among the compelling features of this protein to be used as a pharmaceutical agent. However, there are several impediments to applications of protein pharmaceuticals including: proteolytic degradation, short circulating half-life, low solubility and immunogenicity. Pegylation is a common bioconjugation polymer capable of overcoming these drawbacks. Recombinant expression and purification of calprotectin along with its pegylation would result in enhanced pharmaco-dynamic and pharmacokinetic properties. Our florescence spectroscopy and far Ultraviolet-optical density results indicate that pegylation altered the physical and structural properties of the calprotectin to become in a more stable and functionally active state. Due to enhanced pharmacodynamic and pharmacokinetic properties of the calprotectin via pegylation, this study would pave the way for better in vitro and in vivo validations of calprotectin applications in medical practice. © 2016, Springer Science+Business Media New York