Synthesis and nicotinic acetylcholine receptor affinity of bivalent tropane‐3‐carboxylates

A series of diol di‐(tropane‐3α‐carboxylate) esters and diol di‐(tropane‐3β‐carboxylate) esters were synthesized from 3‐tropene‐3‐carboxylic acid and tropane‐3β‐carboxylic acid, respectively. The bivalent tropane‐3‐carboxylates were evaluated for their ability to inhibit [³H]cytisine binding at rat brain nicotinic acetylcholine receptors (nAChRs). In general the (3β,3β')‐isomers were more potent than (3α,3α')‐isomer and the (3β,3β')‐decyl derivative (n = 10, K_i = 145 nM) exhibited the most potent affinity for nAChRs of the series.     

β-d-Xylosidase from Geobacillus thermoleovorans IT-08: Biochemical Characterization and Bioinformatics of the Enzyme

The gene encoding a thermostable β-d-xylosidase (GbtXyl43B) from Geobacillus thermoleovorans IT-08 was cloned in pET30a and expressed in Escherichia coli; additionally, characterization and kinetic analysis of GbtXyl43B were carried out. The gene product was purified to apparent homogeneity showing M _r of 72 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme exhibited an optimum temperature and pH of 60 °C and 6.0, respectively. In terms of stability, GbtXyl43B was stable at 60 °C at pH 6.0 for 1 h as well as at pH 6–8 at 4 °C for 24 h. The enzyme had a catalytic efficiency (k _cat/K _M) of 0.0048?±?0.0010 s^?1 mM^?1 on p-nitrophenyl-β-d-xylopyranoside substrate. Thin layer chromatography product analysis indicated that GbtXyl43B was exoglycosidase cleaving single xylose units from the nonreducing end of xylan. The activity of GbtXyl43B on insoluble xylan was eightfold higher than on soluble xylan. Bioinformatics analysis showed that GbtXyl43B belonging to glycoside hydrolase family 43 contained carbohydrate-binding module (CBM; residues 15 to 149 forming eight antiparallel β-strands) and catalytic module (residues 157 to 604 forming five-bladed β-propeller fold with predicted catalytic residues to be Asp287 and Glu476). CBM of GbtXyl43B dominated by the Phe residues which grip the carbohydrate is proposed as a novel CBM36 subfamily.     

Investigation of Yeast Invertase Immobilization onto Cupric Ion-Chelated,Porous, and Biocompatible Poly(Hydroxyethyl Methacrylate-<Emphasis Type="Italic">n</Emphasis>-Vinyl Imidazole) Microspheres

Cupric ion-chelated poly(hydroxyethyl methacrylate-n-vinyl imidazole) (poly(HEMA-VIM)) microspheres prepared by suspension polymerization were investigated as a specific adsorbent for immobilization of yeast invertase in a batch system. They were characterized by scanning electron microscopy, surface area, and pore size measurements. They have spherical shape and porous structure. The specific surface area of the p(HEMA-VIM) spheres was found to be 81.2 m²/g with a size range of 70–120 μm in diameter, and the swelling ratio was 86.9%. Then, Cu(II) ion chelated on the microspheres (546 μmol Cu(II)/g), and they were used in the invertase adsorption. Maximum invertase adsorption was 51.2 mg/g at pH 4.5. Cu(II) chelation increases the tendency from Freundlich-type to Langmuir-type adsorption model. The optimum activity for both free and adsorbed invertase was observed at pH 4.5. The optimum temperature for the poly(HEMA-VIM)/Cu(II)-invertase system was found to be at 55 °C, 10 °C higher than that of the free enzyme at 45 °C. V _max values were determined as 342 and 304 U/mg enzyme, for free and adsorbed invertase, respectively. K _m values were found to be same for free and adsorbed invertase (20 mM). Thermal and pH stability and reusability of invertase increased with immobilization.     

Optimization of Ultrasonic—Assisted Extraction (UAE) Method Using Natural Deep Eutectic Solvent (NADES) to Increase Curcuminoid Yield from Curcuma longa L., Curcuma xanthorrhiza,and Curcuma mangga Val.

This study aims to optimize ultrasonic-assisted natural deep eutectic solvents (NADES) based extraction from C. longa. Choline chloride-lactic acid (CCLA-H₂O = 1:1, b/v) was used to investigate the impact of various process parameters such as solvent’s water content, solid loading, temperature, and extraction time. The optimal yield of 79.635 mg/g of C. longa was achieved from extraction in 20% water content NADES with a 4% solid loading in 35 °C temperature for 1 h. Peleg’s model was used to describe the kinetics of the optimized ultrasonic-assisted extraction (UAE) method, and the results were found to be compatible with experimental data. The optimum conditions obtained from C. longa extraction were then used for the extraction of C. xanthorriza and C. mangga, which give yields of 2.056 and 31.322 mg/g, respectively. Furthermore, n-hexane was utilized as an anti-solvent in the separation process of curcuminoids extract from C. longa, C. xanthorriza, and C. mangga, which gave curcuminoid recovery of 39%, 0.74%, and 27%, respectively. Solidification of curcuminoids was also carried out using the crystallization method with n-hexane and isopropanol. However, the solution of CCLA and curcuminoids formed a homogeneous mixture with isopropanol. Hence, the curcuminoids could not be solidified due to the presence of NADES in the extract solution.