This paper reports on novel and efficient enhancement effects of fruit juice aroma using immobilized β-glucosidase, the enzyme involved in important functions in living organisms, onto superparamagnetic nanoparticles Fe3O4 via carbodiimide β-glucosidase was purified from mandarin (Citrus reticulata) using ammonium sulfate precipitation and hydrophobic interaction chromatography. To be used in this study, superparamagnetic nanoparticles were synthesized and then the shape, size, and magnetism properties of the nanoparticles were characterized. The purified enzyme was immobilized on the nanoparticles. The optimum temperature for β-glucosidase (40 ℃) was increased by 10 ℃ after immobilization, while the optimum pH values of free and immobilized β-glucosidase were 5.5. While the Km and Vmax values of the free enzyme were 0.264 mM and 294 EU, immobilized enzyme’s Km and Vmax were 0.222 mM and 370 EU, respectively. In addition, it was determined that the storage stability of the immobilized enzyme was higher than the free enzyme. When the effect of some metal ions on the enzyme activity was examined, it was observed that Fe+2 increased the enzyme activity while other metals inhibited it. According to the results obtained, the immobilized enzyme had a flavor-enhancing effect on mandarin juice.

Aims: Determination of the biochemical properties of β-glucosidase in peppermint, which is rich in aromatic compounds. Study Design: β-glucosidase was purified from mint, and biochemical characterization of the purified enzyme was performed. Place and Duration of Study: This study was carried out in the Faculty of Arts and Sciences Biochemistry laboratory. Methodology: Enzyme purification was performed by hydrophobic interaction chromatography using a Sepharose 4B-L-tyrosine-1-naphthylamine gel. Optimum pH, temperature, and substrate specificity of the purified enzyme were determined. The effects of glucose, δ-gluconolactone and some heavy metals on the enzyme activity were investigated. Results: The enzyme was purified with 8-fold and 28% yield. The purified protein from mint was visualized at 65 kDa on SDS-PAGE. The substrate specificity of the purified β-glucosidase from mint was determined against para- and ortho-nitrophenyl β-D-glucopyranoside (p/o-NPG) substrates. The Km values were 0.4 and 0.9 mM, and the Vmax values were 102.2 EU and 96.6 EU, respectively. While the optimum pH for the purified enzyme was 6, the optimum temperature was 35°C. Effects of heavy metals Ag+2, Fe+3, Zn+2, Cu+2, and Pb+2 on the purified enzyme activity were investigated. Relative activities of heavy metals were introduced into the reaction medium as 0.75 mM samples without any known inhibitors in the environment. Fe+3 increased the enzyme activity, and Ag+2, Pb+2, Cu+2, and Zn+2 inhibited the enzyme, and their relative activities were 78, 76, 22, and 31%, respectively. Glucose and δ-gluconolactone competitively inhibited the enzyme activity when p-NPG was the substrate. Ki values of glucose and δ-gluconolactone were determined as 0.034±0.001 and 0.038±0.002 mM, respectively. Conclusion: Determination of the biochemical properties of β-glucosidase from mint, which has commercial and pharmacological importance due to the phenolic substances it contains, will contribute to studies on improving food quality.

Leukocytes are isolated by centrifugation after specific lysis of erythrocytes

β-Glucosidase was purified from Brassica oleracea by salting out with ammonium sulfate and hydrophobic interaction chromatography. Results demonstrated that the enzyme is a dimer (130 kD) made up of one major (80 kD) and one minor subunit (50 kD). The pH optimum is 6.0, with 50% of the enzyme's original activity remaining between pH 4.0 and pH 7.0. The temperature optimum is 35C, and activity did not decrease after two hours of exposure to this temperature. The activity of the enzyme was investigated on four substrates, 4-Nitrophenyl β-D-glucopyranoside (p-NPG), ortho-Nitrophenyl-β-D-glucopyranoside (o-NPG), para-Nitrophenyl-β-D-galactoside (p-NPGal) and ortho-Nitrophenyl-β-D-galactoside (o-NPGal), and km values were shown to be 0.755 mM, 0.174 mM, 0.988 mM and 0.213 mM, while Vmax values were 604 U/mg, 38 U/mg, 556 U/mg and 308 U/mg, respectively. The enzyme is completely inhibited by gluconolactone and glucose against p-NPG as substrate, with ki values of 0.038 mM and 0.64 mM, respectively. To our knowledge, this is the first study demonstrating purification and characterization of β-glucosidase from broccoli, thus providing a better understanding of its role in the plant, and establishing a basis for further research. Practical Applications To our knowledge, this is the first study demonstrating purification and characterization of β-glucosidase from broccoli, thus providing a better understanding of its role in the plant, and establishing a basis for further research. The results of this research highlight the potential of the enzyme isolated from broccoli for further research. Succeeding efforts would involve optimization of this procedure for increasing the enzyme yield, in order to make it a viable candidate for industrial application.

Gaucher disease is a hereditary genetic abnormality which defects the pathway of sphingolipid catabolism. The mutation of GBA gene which encodes lysosomal β-glucosidase enzyme is the main characteristics of the disease also is observed in different cancer types. To find the relation between the disease and colon adenocarcinoma, the responsible gene expression of Gaucher disease was analyzed. The gene expression of colon adenocarcinoma was was compared between death and alive patients and analyzed statistically to profile the differences between Gaucher disease genes expression changes. GBA, GBA2, GBA3, SCARB2 and PSAP have the maximum genetic alteration which is observed in colon adenocarcinoma.

Analysis of Y-chromosome haplogroup distribution is widely used when investigating geographical clustering of different populations, which is why it plays an important role in population genetics, human migration patterns and even in forensic investigations. Individual determination of these haplogroups is mostly based on the analysis of single nucleotide polymorphism (SNP) markers located in the non-recombining part of Y-chromosome (NRY). On the other hand, the number of forensic and anthropology studies investigating short tandem repeats on the Y-chromosome (Y-STRs) increases rapidly every year. During the last few years, these markers have been successfully used as haplogroup prediction methods, which is why they have been used in this study. Previously obtained Y-STR haplotypes (23 loci) from 100 unrelated Turkish males recently settled in Sarajevo were used for the determination of haplogroups via 'Whit Athey's Haplogroup Predictor' software. The Bayesian probability of 90 of the studied haplotypes is greater than 92.2% and ranges from 51.4% to 84.3% for the remaining 10 haplotypes. A distribution of 17 different haplogroups was found, with the Y- haplogroup J2a being most prevalent, having been found in 26% of all the samples, whereas R1b, G2a and R1a were less prevalent, covering a range of 10% to 15% of all the samples. Together, these four haplogroups account for 63% of all Y-chromosomes. Eleven haplogroups (E1b1b, G1, I1, I2a, I2b, J1, J2b, L, Q, R2, and T) range from 2% to 5%, while E1b1a and N are found in 1% of all samples. Obtained results indicate that a large majority of the Turkish paternal line belongs to West Asia, Europe Caucasus, Western Europe, Northeast Europe, Middle East, Russia, Anatolia, and Black Sea Y-chromosome lineages. As the distribution of Y-chromosome haplogroups is consistent with the previously published data for the Turkish population residing in Turkey, it was concluded that the analyzed population could also be recognized as a representative sample of the Turkish population residing in Turkey.

Forensic botany refers to the application of plant biology disciplines in crime investigations. Since plants are widespread in most habitats, they often provide valuable clues in criminal investigations. Hence, the primary role of forensic botany is to establish a connection between the victim or suspect and the crime scene. The forensic applications of forensic botany include many aspects including recognition of plant evidence at the crime scene, collection and preservation of plant evidence, maintenance of the chain of custody, and a testimony for the admissibility of this evidence in court. For the development of new efficient methods for plant identification and individualization, the initial step is optimization of DNA isolation from plant material. The aim of this work is to optimize the DNA isolation protocol of different leaf and non-leaf plant tissues, including pollen grains, using the salting out method from Taraxacum officinale, a species of the Asteraceae family. Since the species is very common in ecosystems with antropogenic influence, the need to optimize the DNA isolation protocol is of high importance. The efficiency of the isolation protocol was evaluated using spectrophotometry (concentration and purity) and gel electrophoresis. The results demonstrate that salting out can be used for the isolation of DNA from Taraxacum officinale. However, optimization is required, depending on the part of the plant from which DNA is extracted.

Mixed lineage leukemias (MLL) express unique clinical and biological characteristics. MLL interacts with over 50 different genes resulting in expression of chimeric proteins whereby the MLL amino-terminal portion is fused to the carboxy-terminal portion of the partner. Chromosomal translocations t(9 ; 11), t(11 ; 19) and t(4 ; 11) leading to MLL-AF9, MLL-ENL and MLL-AF4 fusions are the most frequent. Leukemias with MLL rearrangements are mostly driven through dysregulation of a transforming gene expression program, making it a unique epigenetic model of leukemia. Using the whole genome profiling of acute myeloid leukemias (AMLs), we analysed the differential gene expression, methylation pattern, and mutational spectra between MLL and other AML types (n=197). We found that 120 genes were differentially expressed, with 36 genes characterized by more than twofold expression difference. In this study, we will present differently expressed genes with and their potential role in leukemogenesis. Since rearrangements of the MLL gene lead to aberrant methylation, we investigated differential methylation patterns among MLL and other AML types and identified affected methylation hotspots.

Abstract Inhibitory effects of some synthesized dihydroxycoumarin compounds on purified G6PD were investigated. For this purpose, initially human erythrocyte G6PD was purified 7069-fold in a yield of 33.6% by using ammonium sulfate precipitation and affinity chromatography which includes 2′,5′-ADP Sepharose 4B. The purified enzyme showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Enzyme activity was determined spectrophotometrically according to Beutler method at 340 nm. 6,7-Dihydroxy-3-(2-methylphenyl)-2H-chromen-2-one (OPC), 6,7-dihydroxy-3-(3-methylphenyl)-2H-chromen-2-one (MPC) and 6,7-dihydroxy-3-(4-methylphenyl)-2H-chromen-2-one (PPC) were used as dihydroxycoumarin compounds. This study has demonstrated that G6PD activity is very highly sensitive to study coumarin derivatives.

The greater wax moth, Galleria mellonella, is one of the most ruinous pests of honeycomb in the world. Beta-glucosidases are a type of digestive enzymes that hydrolytically catalyzes the beta-glycosidic linkage of glycosides. Characterization of the beta-glucosidase in G. mellonella could be a significant stage for a better comprehending of its role and establishing a safe and effective control procedure primarily against G. mellonella and also some other insect pests. Laboratory reared final instar stage larvae were randomly selected and homogenized for beta-glucosidase activity assay and subsequent analysis. The enzyme was purified to apparent homogeneity by salting out with ammonium sulfate and using sepharose-4B-l-tyrosine-1-naphthylamine hydrophobic interaction chromatography. The purification was 58-fold with an overall enzyme yield of 29%. The molecular mass of the protein was estimated as ca. 42 kDa. The purified beta-glucosidase was effectively active on para/ortho-nitrophenyl-beta-d-glucopyranosides (p-/o-NPG) with Km values of 0.37 and 1.9 mM and Vmax values of 625 and 189 U/mg, respectively. It also exhibits different levels of activity against para-nitrophenyl-β-d-fucopyranoside (p-NPF), para/ortho-nitrophenyl β-d-galactopyranosides (p-/o-NPGal) and p-nitrophenyl 1-thio-β-d-glucopyranoside. The enzyme was competitively inhibited by beta-gluconolactone and also was very tolerant to glucose against p-NPG as substrate. The Ki and IC50 values of δ-gluconolactone were determined as 0.021 and 0.08 mM while the enzyme was more tolerant to glucose inhibition with IC50 value of 213.13 mM for p-NPG.