Sharif Digital Repository

Asparagainase is a therapeutic enzyme which has been a subject of research for decades. The enzyme catalyzes the hydrolysis of the amide group in asparagine and similar amides. Altering the substrate specificity and stabilization of this enzyme can increase its therapeutic properties. Moreover, asparaginases may be evolved to catalyze the hydrolysis of other similar compounds. These can be achieved through directed evolution and computational methods.In this study, the gene encoding L-asparaginase II enzyme from E. coli was amplified by polymerase chain reaction (PCR) and was cloned into an expression vector. The recombinant protein was expressed by an appropriate host secreting the... 

Separation of biomolecules such as enzymes is the most important part of biotechnology process which 10-90 % of costs is for separation and purification. There are several methods for purification of bio materials and one of them is chromatography which use in enzyme purification but this method have several problem such as cost ,time consuming and scale up Because of this a suitable alternative demands for industrial separation. An alternative is aqueous two-phase systems (ATPS). In this project was studied about partial purification of alkaline protease produced by Bacillus licheniformis ATCC 21424 in aqueous two-phase system and comparison with chromatography method. an aqueous two-phase... 

Osteoarthritis has always been one of the most common diseases in middle age because it causes severe pain and inflammation in the joints of the body. Cartilage tissue does not have the ability to repair itself. For this reason, fabricating and designing the most efficient, least-expensive, and most convenient methods for the treatment of cartilage defects is always an important issue. Today, there are surgical and injectable methods to relieve pain and initiate the body's natural healing response. But due to the many disadvantages and limitations of these methods, tissue engineering science has turned to modifying these methods or providing new methods. One of these methods is the use of... 

Pharmaceutical active compounds existence in the water would cause serious ecological risks and human health-related adverse effects which turn to environmental concern, therefore several studies have done to remove pharmaceutical active compounds efficiently. Carbamazepine (CBZ), a widely used psychiatric drug, is one of the most frequently detected compounds in the surface water and groundwater which is studies in the current study. Recently, biocatalytic degradation using ligninolytic enzymes such as laccase provides a promising approach for their removal from water and wastewater. In this work, carbamazepine removal by immobilized laccase on modified membrane by multi wall carbon nanotube... 

Fats and oils are one of the main human food diet components that provide the necessary fatty acids for our body. So the fatty acids available in consuming oils are so important. In this research the mixture of palm and sunflower oils with different compounds: (80:20, 60:40, 50:50, 40:60, 20:80) were used for preparing free- trans shortening by interesterification. For this purpose a special stabilized lipase (Lipozyme TLIM) were used and performed the physicochemical properties: free fatty acids, peroxide value, iodine value, slip melting point, solid fat content, P-anisidine value, soap, oxidative stability at 110 0C, and the profile of fatty acids on the primitive blends and final samples... 

Dietary CLA production for medicinal purposes needs a safe and selective process. Biological production methods meet this goal. In this study, the microbial species that was suitable for CLA production was selected; then the CLA produced by bioconversion methods was utilized to produce CLA-enriched triglycerides by trans-esterification. Also the biochemical kinetics of CLA production and esterification reactions to produce CLA-enriched triglycerides was studied. In the presence of two substrates, the kinetics of enzymatic reaction was studied. Then the factors influencing on the microbial yield and CLA production including reaction conditions (reaction time, cell concentration and substrate... 

In this study, the polysulfon ultrafilteration membranes were prepared and applied to concentrate the Lipase enzymes produced in the laboratory and the commercial enzymes of Novozymes Company. Using Design Expert software. The effect of membrane preparation conditions including polysulfone and polyvinylpyrrolidone concentration and the solvent type were investigated. The prepared membranes were characterized by measuring of distilled water flux, enzyme liquor flux, and enzyme activity in permeate and the Retentate flows. The results showed that increasing of polysulfone concentration reduced the flux of distilled water and enzyme activity in permeate and enhanced the enzyme activity in the... 

Enzymes are biological catalysts that reduce the activation energy of a reaction in a short time and thus increase the speed of the reaction. The most common enzymes used in various industries, including the detergent industry, are lipases, amylases, celluloses, and proteases. Proteases are one of the most important industrial enzymes. The performance of enzymes is affected by the durability of their activity in long-term storage. Therefore, using a technique that can maintain the activity of enzymes, under storage and usage conditions, is an essential step in making a product containing enzymes. Stabilization-immobilization operation is one of the important downstream processes in... 

In this research, the amount of partitioning Invertase to be evaluated in nine systems, these systems contain Peg 2000,Peg 4000 & Peg 6000 with three different salt.In first, Binodal curve for nine systems to be drown by cloudy point method, then five point be selected in each system such as sample feed. Invertase in every systems to be solved in constant weight fraction. The analysis of every phasesto be achieved by Merchuk method. For correlation data of this method to be used by Othmer-Tobias method and then using equation Mrchvk ensure the accuracy of phase analysis , analysis of the results obtained with the UV method was studied.Accuracy of UV analysis was studied by using lattice... 

Conjugation of proteins to nanoparticles has numerous applications in sensing, imaging, delivery, catalysis, therapy and control of protein structure and activity. Therefore, characterizing the nanoparticle–protein interface is of great importance. A variety of covalent and non-covalent linking chemistries have been reported for nanoparticle attachment. Typically, a specific protein residue is linked directly to the nanoparticle core or to the ligand. As conjugation often affects the protein structure and function, techniques to probe structure and activity are assessed Characterization studies of nanoparticle–protein complexes show that the structure and function are influenced by the... 

Superparamagnetic core/shell nonporous (S1) and core/shell/shell mesoporous (S2 and S3) nanocomposite magnetite/silica particles with a magnetite cluster core of 130±30 nm, a nonporous silica shell of 90±10 nm thickness, and a mesoporous silica shell of 70±15 nm thickness were prepared thorough a simple method. Mesoporous particles were prepared with two BJH pore sizes (2.44 for S2 and 3.76 nm for S3 particles, respectively). The fabricated S1, S2 and S3 particles present high saturation magnetization values of 20, 13 and 17 emu/g, respectively. As a biological application, the lipase from Pseudomonas cepacia was successfully immobilized onto the amino-functionalized nanocomposite particles... 

Using free laccase, enzymatic decolorization of two azo dyes were optimized. The optimum conditions with the aim of maximizing the decolorization of AB92 dye (92.30%) were: dye concentration = 11.85 mg L-1, pH = 5.1, and enzyme concentration = 98.37 mg L-1. While, these conditions to achieve the maximum decolorization yield of DR23 (95.60%) were: dye concentration = 17.68 mg L-1, pH 3.7 and enzyme concentration =97.89 mg L-1.In the second part, the laccase enzyme was covalently immobilized onto GO nanosheets. At the concentration of graphene oxide and laccase enzyme equal to 1 mg mL-1 and 0.9 mg mL-1, respectively, the enzyme loading was 156.5 mg g-1 and the immobilization efficiency was... 

Biodiesel has drawn the attention of many researches to itself as a replacing source for crude oil, recently. Transesterification is one of the most popular methods used for biodiesel production. Enzyme transesterification is a more simple method in comparison with acidic or basic transesterification. However, the high price and low yield of enzyme transesterification has caused many obstacles in its application. Enzyme engineering has gained much attention as a way to improve enzyme performance. In this thesis, biodiesel production by a native enzyme named Bacillus thermocatenulatus, and two engineered-enzymes is studied. First, effective parameters and variables in biodiesel production by... 

Industrial enzymes can generally be classified into three groups: carbohydrases, proteases and lipases, which account for a major part of the global enzyme trade. Proteases, in particular, have always been extensively studied and investigated due to their widespread use in various industries, such as food, pharmaceutical, chemical, and agricultural sectors. Proteases are a group of proteolytic enzymes that break down proteins and peptides into their constituent amino acids by hydrolyzing peptide bonds. Among the family of proteases, alkaline serine proteases have special biocatalytic abilities in both aqueous and organic environments. Their suitable resistance in alkaline environments and... 

In this research, the partitioning of Penicillin G-Acylase in aqueous two-phase systems (ATPS) containing poly ethylene glycol (PEG) 20000 or 35000 and potassium di-hydrogen phosphate (KH2PO4) or sodium citrate (C6H5Na3O7.5H2O) is measured at three temperature (301.2, 307.2 and 310.2) K. The effects of temperature, polymer molecular weight, polymer and salt concentrations on partitioning of Penicillin G in the aqueous two-phase systems are studied. The experimental data showed that composition of salt has a large effect on partitioning of Penicillin G in ATPS and temperature of system has small effect on the partitioning. The modified local composition model was used to obtain the activity... 

β-galactosidase enzyme is immobilized from Kluyveromyces lactis by Microencapsulation method in various compositions of sodium alginate and Carboxymethyl cellulose. Optimum stabilization was gained at 1.85% sodium alginate, 0.9% carboxymethyl cellulose and 2% calcium chloride through applying Surface response methodology. Yield of immobilization and activity was 54.94% and 65.27%. variation of yield of immobilization versus alginate concentration was a quadratic curve with the maximum. When alginate's concentrations is low, the cavities diameter is great, so the enzyme leaks from capsule while immobilizing. At high concentrations of alginate, despite the smaller cavities, immobilization... 

Nowadays , application of enzymes as biocatalysts in wastewater treatment processes is increasing. It is economically important to reuse enzymes and increase their stability in these types of processes. Therefore, was immobilization of enzymes and using them in wastewater treatment have a great importance.In this project, Horseradish peroxidase enzyme, whose effect on phenol removal reactions in other studies has been proven before, was immobilized on the alginate-coated magnetic nanoparticles, to investigate the effect of immobilization on the stability and function of this enzyme for phenol removal. The enzyme was immobilized at the temperature of 25 ° C and pH = 8, according to the data... 

L-asparaginase II is an amidohydrolase which hydrolyzes L-asparagine to furnish L-aspartic acid and ammonia. The enzyme has played an important role for 40 years in treating cancer in persons with juvenile leukemias or lymphomas. Furthermore, the enzyme has become important in food industries and biosensors technology.In this study, the expression of recombinant L-asparaginase II in E. Coli was optimized and the purification of this enzyme from growth culture was carried out with nickel-charged affinity resin. Afterwards, the activity, and kinetic parameters of purified recombinant L-asparaginase II including Km, kcat, Vmax, and enzyme efficiency were measured.In the next step, random... 

After proteins, one of the important biological macromolecules, are synthesized by ribosomes,they automatically reach their own special 3-D structure. This native structur or better known as folded state is what determines the function of a protein. In some situations, the proteins become misfolded leading to its lack of biological function through aggregation or fibril formation known as "amyloid". The amyloid fibrils have been shown to be a causative factor in nerve decay such as Alzheimer's. Therefore, inhibition of amyloid formation using different approaches such as synthetic organic and natural compounds has, recently, been under spotlight. A pyridazine basedcompound (RS-0406) has... 

L-asparaginase enzyme is a well known enzyme due to itschemotherapeutic properties. However, this is not the only feature of the enzyme. L- asparaginase can be also used in food processing to reduce of acrylamide which is a carcinogenic substance. furthermore, acrylamide formation, is prevented by addition of the enzyme before cooking process of some foods such as chips, which contains sugar and asparagine amino acid. Due to use of the enzyme in food industry, we need to produce enzyme with high activity and cheaper raw material. In the present study, Candida utilis ATCC 9950 was used to produce asparaginase enzyme, and beet molasses and yeast extract was used as a carbohydrate and...