Scholarly

Effect of Anion and Amino Functional Group on Resin for Lipase Immobilization with Adsorption-Cross Linking Method

Year: 2015 Volume: 9 Issue: 12 1518 - 1522 Pages

Abstract: Lipase is one of biocatalyst which is applied commercially for the process in industries, such as bioenergy, food, and pharmaceutical industry. Nowadays, biocatalysts are preferred in industries because they work in mild condition, high specificity, and reduce energy consumption (high pressure and temperature). But, the usage of lipase for industry scale is limited by economic reason due to the high price of lipase and difficulty of the separation system. Immobilization of lipase is one of the solutions to maintain the activity of lipase and reduce separation system in the process. Therefore, we conduct a study about lipase immobilization with the adsorption-cross linking method using glutaraldehyde because this method produces high enzyme loading and stability. Lipase is immobilized on different kind of resin with the various functional group. Highest enzyme loading (76.69%) was achieved by lipase immobilized on anion macroporous which have anion functional group (OH‑). However, highest activity (24,69 U/g support) through olive oil emulsion method was achieved by lipase immobilized on anion macroporous-chitosan which have amino (NH2) and anion (OH-) functional group. In addition, it also success to produce biodiesel until reach yield 50,6% through interesterification reaction and after 4 cycles stable 63.9% relative with initial yield. While for Aspergillus, niger lipase immobilized on anion macroporous-kitosan have unit activity 22,84 U/g resin and yield biodiesel higher than commercial lipase (69,1%) and after 4 cycles stable reach 70.6% relative from initial yield. This shows that optimum functional group on support for immobilization with adsorption-cross linking is the support that contains amino (NH2) and anion (OH-) functional group because they can react with glutaraldehyde and binding with enzyme prevent desorption of lipase from support through binding lipase with a functional group on support.

Preparation of Low-Molecular-Weight 6-Amino-6-Deoxychitosan (LM6A6DC) for Immobilization of Growth Factor

Year: 2015 Volume: 9 Issue: 3 296 - 299 Pages

Abstract: Epidermal Growth Factor (EGF, Mw=6,045) has been reported to have high efficiency of wound repair and anti-wrinkle effect. However, the half-life of EGF in the body is too short to exert the biological activity effectively when applied in free form. Growth Factors can be stabilized by immobilization with carbohydrates from thermal and proteolytic degradation. Low molecular weight chitosan (LMCS) and its derivate prepared by hydrogen peroxide has high solubility. LM6A6DC was successfully prepared as a reactive carbohydrate for the stabilization of EGF by the reactions of LMCS with alkalization, tosylation, azidation and reduction. The structure of LM6A6DC was confirmed by FT-IR, 1H NMR and elementary analysis. For enhancing the stability of free EGF, EGF was attached with LM6A6DC by using water-soluble carbodiimide. EGF-LM6A6DC conjugates did not show any cytotoxicity on the Normal Human Dermal Fibroblast (NHDF) 3T3 proliferation at least under 100 μg/ml. In the result, it was considered that LM6A6DC is suitable to immobilize of growth factor.

Immobilization of Lipase Enzyme by Low Cost Material: A Statistical Approach

Year: 2014 Volume: 8 Issue: 6 588 - 591 Pages

Abstract: Immobilization of lipase enzyme produced from palm oil mill effluent (POME) by the activated carbon (AC) among the low cost support materials was optimized. The results indicated that immobilization of 94% was achieved by AC as the most suitable support material. A sequential optimization strategy based on a statistical experimental design, including one-factor-at-a-time (OFAT) method was used to determine the equilibrium time. Three components influencing lipase immobilization were optimized by the response surface methodology (RSM) based on the face-centered central composite design (FCCCD). On the statistical analysis of the results, the optimum enzyme concentration loading, agitation rate and carbon active dosage were found to be 30 U/ml, 300 rpm and 8 g/L respectively, with a maximum immobilization activity of 3732.9 U/g-AC after 2 hrs of immobilization. Analysis of variance (ANOVA) showed a high regression coefficient (R2) of 0.999, which indicated a satisfactory fit of the model with the experimental data. The parameters were statistically significant at p

Synthesis of Peptide Amides using Sol-Gel Immobilized Alcalase in Batch and Continuous Reaction System

Year: 2011 Volume: 5 Issue: 4 369 - 374 Pages

Abstract: Two commercial proteases from Bacillus licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were screened for the production of Z-Ala-Phe-NH2 in batch reaction. Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal amidation of Z-Ala-Phe-OMe using ammonium carbamate as ammonium source. Immobilization of protease has been achieved by the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and tetramethoxysilane (TMOS) as precursors (unpublished results). In batch production, about 95% of Z-Ala-Phe-NH2 was obtained at 30°C after 24 hours of incubation. Reproducibility of different batches of commercial Alcalase 2.4 L FG preparations was also investigated by evaluating the amidation activity and the entrapment yields in the case of immobilization. A packed-bed reactor (0.68 cm ID, 15.0 cm long) was operated successfully for the continuous synthesis of peptide amides. The immobilized enzyme retained the initial activity over 10 cycles of repeated use in continuous reactor at ambient temperature. At 0.75 mL/min flow rate of the substrate mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5 hours of substrate recycling. The product contained about 90% peptide amide and 10% hydrolysis byproduct.

A Novel Strategy for Oriented Protein Immobilization

Year: 2009 Volume: 3 Issue: 5 255 - 259 Pages

Abstract: A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.

Dextran Modified Silicon Photonic Microring Resonator Sensors

Year: 2012 Volume: 6 Issue: 9 434 - 437 Pages

Abstract: We present a dextran modified silicon microring resonator sensor for high density antibody immobilization. An array of sensors consisting of three sensor rings and a reference ring was fabricated and its surface sensitivity and the limit of detection were obtained using polyelectrolyte multilayers. The mass sensitivity and the limit of detection of the fabricated sensor ring are 0.35 nm/ng mm-2 and 42.8 pg/mm2 in air, respectively. Dextran modified sensor surface was successfully prepared by covalent grafting of oxidized dextran on 3-aminopropyltriethoxysilane (APTES) modified silicon sensor surface. The antibody immobilization on hydrogel dextran matrix improves 40% compared to traditional antibody immobilization method via APTES and glutaraldehyde linkage.

Heat Treatment and Rest-Inserted Exercise Enhances EMG Activity of the Lower Limb

Year: 2007 Volume: 1 Issue: 11 150 - 153 Pages

Abstract: Prolonged immobilization leads to significant weakness and atrophy of the skeletal muscle and can also impair the recovery of muscle strength following injury. Therefore, it is important to minimize the period under immobilization and accelerate the return to normal activity. This study examined the effects of heat treatment and rest-inserted exercise on the muscle activity of the lower limb during knee flexion/extension. Twelve healthy subjects were assigned to 4 groups that included: (1) heat treatment + rest-inserted exercise; (2) heat + continuous exercise; (3) no heat + rest-inserted exercise; and (4) no heat + continuous exercise. Heat treatment was applied for 15 mins prior to exercise. Continuous exercise groups performed knee flexion/extension at 0.5 Hz for 300 cycles without rest whereas rest-inserted exercise groups performed the same exercise but with 2 mins rest inserted every 60 cycles of continuous exercise. Changes in the rectus femoris and hamstring muscle activities were assessed at 0, 1, and 2 weeks of treatment by measuring the electromyography signals of isokinetic maximum voluntary contraction. Significant increases in both the rectus femoris and hamstring muscles were observed after 2 weeks of treatment only when both heat treatment and rest-inserted exercise were performed. These results suggest that combination of various treatment techniques, such as heat treatment and rest-inserted exercise, may expedite the recovery of muscle strength following immobilization.

Top Journal