Enzyme reactions

Most of the health benefits of fish oil can be attributed to the presence of omega-3 fatty acids like Docosahexenoic acid (DHA) and Eicosapentaenoic acid (EPA). There are few dietary sources of EPA and DHA other than oily fish. EPA and DHA have great potential effect on human health. In this research, Supercritical carbon dioxide (scCO 2) extracted mackerel oil was reacted by enzyme at different systems to improve the EPA and DHA. Different types of immobilize enzyme TL-IM, RM-IM, Novozyme 435 were assessed for improving PUFAs. Best result was found at nonpressurized system using TL-IM. Reacted oil particle were obtained with polyethylene glycol by gas saturated solution process (PGSS). Different parameters like temperature, pressure, agitation speed and nozzle size effect on particle formulation were observed. SEM and PSA analysis showed, small size non spherical particles were obtained. It was found that after particle formation poly unsaturated fatty acids (PUFAs) were present in particle as same in oil. PUFAs release from particle was almost linear against constant time duration. Oil quality in particle not change significantly, in this contrast this study will be helpful for food and pharmaceutical industry to provide high EPA and DHA containing powder. Keywords Supercritical CO 2. Omega-3 fatty acids. Immobilize enzyme. Particle. Control release Research Highlights • Supercritical CO 2 extracted mackerel oil was used • 3 different immobilized lipase enzymes were used to improve polyunsaturated fatty acid. • Gas saturated solution (PGSS) process was used for particle formation • Different parameters were observed for optimization in PGSS process • Control release were observed of PUFAs from particle

Wheat germ oil is a unique source of highly concentrated nutrient; its beneficial active ingredients could have a substantial impact on human health considering the amount that is used in cooking. In this study, wheat germ oil was micronized with polyethylene glycol using particles from gas-saturated solutions (PGSS) process and investigated functional materials content in particle changing different parameters. PGSS was carried out at temperatures of 40-50C, pressures of 10-30 MPa and at different ratios (10:1, 5:1). The average particle size was found from 0.6330 ± 0.004 to 1.3506 ± 0.011 μm at a ratio of 10:1 and from 1.1776 ± 0.011 to 3.7680 ± 0.016 μm at a ratio of 5:1. In fatty acid compositions, linoleic acid was present and had the highest amount in all conditions after particle formation. Total phenolic compound and antioxidant capacity were also found in particles. PRACTICAL APPLICATION Wheat germ is mainly produced as a by-product in the wheat milling industry. This by-product is generally used as animal feed; however, it contains oil that contains high amounts of functional materials like polyunsaturated fatty acids and phenolic compounds that have practical uses in life. These have great health benefits for humans and can be used in products such as foods, biological insect control agents, pharmaceuticals and cosmetic formulations. But for protection from oxidation, preservation of bioactivity and because of control release and dose measurement, particle formation is the best way. Small particles of pharmaceuticals with a narrow particle size distribution play a vital role in the design of conventional drug delivery systems like tablets, capsules, injections and biphasic drug delivery systems.

Fucoxanthin is a precious marine carotenoid with multiple bioactivities such as antioxidant, anti-obesity, and anti-cancer effects. Despite its valuable properties, fucoxanthin possesses high sensitivity when exposed to aerial, thermal, or light factors. Our study aims to extract fucoxanthin-rich oil (FO) from Saccharina japonica using supercritical carbon dioxide with sunflower oil (SFO) as co-solvent and encapsulate the extracted oil using particles from gas-saturated solutions (PGSS) process with polyethylene glycol (PEG) as a biodegradable coating. The encapsulation efficiency (EE) of the extracted oil based on fucoxanthin content was optimized using response surface methodology with Box-Behnken design (BBD). Three independent factors, including mixing ratio of FO and PEG, temperature, and pressure, were optimized. The encapsulated particles (EP) at optimized conditions were characterized for their powder properties and storage stability. FO and EP were measured in antioxidant activities and fatty acids content. The optimum EE was approximately 82%, implying the capability of PGSS and PEG in encapsulating nutritious oil.

The separation of oil by a suitable technique from the Pacific oyster muscle is important for the utilization of the oil as a ω-3 polyunsaturated fatty acids (ω-3 PUFAs) source and production of bio-functional peptides/ oligosaccharides from oil-free residue. This study was conducted to prepare ω-3 PUFAs concentrate from supercritical carbon dioxide (SC-CO 2) extracted Pacific oyster oil by enzymecatalyzed ethanolysis reactions. SC-CO 2 extractions were done at different temperatures and pressures to optimize suitable extraction conditions and extracted oils were compared with Soxhlet (n-hexane) extracted oil to evaluate the yield and quality. Oil extracted by SC-CO 2 at optimized conditions was used for ethanolysis reaction catalyzed by immobilized sn-1,3 specific lipases, namely Novozymes-435, Lipozyme TLIM, and Lipozyme RMIM to produce 2-monoacylglycerols (2-MAG) rich in ω-3 PUFAs. The optimum temperature and pressure for SC-CO 2 extractions of oyster oil was 50°C and 30 MPa. In this condition, the yield of oil was 5.96% and the acid, peroxide, free fatty acid, and p-anisidine values were 4.49 mg KOH/g, 4.72 meq/kg, 3.42%, and 10.03, respectively. The ω-3 PUFAs content significantly increased in 2-MAG obtained from Novozymes 435, Lipozyme TLIM, and Lipozyme RMIM to 43.03 ± 0.36, 45.95 ± 0.29, and 40.50 ± 0.77%, respectively (p < 0.05). A thin layer chromatography (TLC) analysis confirmed the production and separation of 2-MAG in the ethanolysis process. The ratio of total ω-3 to ω-6 fatty acids was almost twice in 2-MAG of SC-CO 2 extracted oyster oil. SC-CO 2 extracted Pacific oyster oil can be used for sn-1,3 specific lipases catalyzed ethanolysis to produce ω-3 PUFAs rich in 2-MAG.

A relatively complex network of reactions has been investigated, using as a network model the isothermal batch esterification of acetic acid with ethanol in n-heptane catalyzed by lyophilized mycelium of Aspergillus oryzae. The kinetic analysis was firstly carried out on the whole system, without any simplification, by means of the well-known integral method. Owing to the poor results obtained by this way, we developed an alternative approach, combining initial rates and integral analysis and reducing the number of empirical parameters to be determined by the use of equilibrium data. All the values of the parameters calculated according to this "composite" approach to kinetic analysis well correlate with experimental data.

Production of omega-3 fatty acids from lemuru fish by-products was studied by enzymatic hydrolysis using a lipase enzyme in one liter of the batch reactor. The hydrolysis temperature of fish oil was set at 45 to 55 ℃ for 0 to 24 h, whereas agitation from 50 to 150 rpm. RSM-Box Bhenken was used to study the effect of these parameters on omega-3 (EPA, docosahexaenoic acid (DHA), and α-linolenic acid (ALA)) content. The % free fatty acid (FFA), acid index, peroxide index, iodine index, and saponification index of lemuru fish oil was 0.925, 2.52, 42.5, 97.28, and 160.11%, respectively. GC-MS analysis results showed that unsaturated fatty acids content (62.34%), which are consisted of omega-3 (EPA, DHA, and ALA), omega-6 and omega-9, was much higher than saturated acids (12.97%). The experiment data showed that the highest EPA (1.221%) and DHA (0.312%) content were reached at 50 ℃ and 24 h with 150 rpm of agitation. However, through the RSM-Box Bhenken analysis and 3D surface plot, it wa...

Particle design is presently a major development of supercritical fluids applications, mainly in the paint, cosmetic, pharmaceutical, and specialty chemical industries. Supercritical fluids have been successfully used to obtain composites or encapsulates, which comprise an active compound loaded into a matrix of a carrier material, in order to improving product preservation as well as controlling the dissolution rate of the active compound. The particles from gas-saturated solution (PGSS) process has been used to micronize suspensions of different substances in polymer melts, in order to obtain composite materials. The particle formation of functional pigment with biodegradable polymer, polyethelene glycol (PEG) was performed by supercritical carbon dioxide (SCO 2) in a thermostated stirred vessel. PGSS were carried out in different temperatures and pressures to measure the optimum condition for the formation of functional pigment particle (pressure from 20 MPa to 25 MPa, temperature from 40 to 50°C). The average diameter of the particles obtained at different conditions was about 0.8-1.4 µm accompanied by the size reduction, comparing the size of unprocessed particles, 430 µm.

The combined Electron Localization Funtion (ELF)/ Noncovalent Interaction (NCI) topological analysis (Gillet et al. J. Chem. Theory Comput. 2012, 8, 3993) has been extended to enzymatic reaction paths. We applied ELF/NCI to the reactions of DNA polymerase λ and the ε subunit of DNA polymerase III. ELF/NCI is shown to provide insights on the interactions during the evolution of enzymatic reactions including predicting the location of TS from structures located earlier along the reaction coordinate, differential metal coordination, and on barrier differences with two different cations.

Substrate and product concentration data obtained by simulating enzyme-substrate reaction rate equations were used to test two proposed kinetic rate constant estimation techniques in this study. In the first technique, the turnover number, k3, was calculated using early transient time domain data, which are difficult to obtain experimentally. The technique used an iterative approach to calculate k3 with a pair of data and the value of k3 could be retrieved with 35% error. The second technique calculated k3 using stationary domain data and the value of k3 could be retrieved with less than 5% error. This second technique also offered internal consistency in the calculation of k3 by calculating k3 both from the intercept and the slope of the linear plot derived in this study. A series of sensitivity analyses was conducted to understand the robustness of the second technique in estimating k3 from simulated data to the changes in the reaction rate constants (k1, k2, and k3) and the initi...

Production of omega-3 fatty acids from lemuru fish by-products was studied by enzymatic hydrolysis using a lipase enzyme in one liter of the batch reactor. The hydrolysis temperature of fish oil was set at 45 to 55 ℃ for 0 to 24 h, whereas agitation from 50 to 150 rpm. RSM-Box Bhenken was used to study the effect of these parameters on omega-3 (EPA, docosahexaenoic acid (DHA), and α-linolenic acid (ALA)) content. The % free fatty acid (FFA), acid index, peroxide index, iodine index, and saponification index of lemuru fish oil was 0.925, 2.52, 42.5, 97.28, and 160.11%, respectively. GC-MS analysis results showed that unsaturated fatty acids content (62.34%), which are consisted of omega-3 (EPA, DHA, and ALA), omega-6 and omega-9, was much higher than saturated acids (12.97%). The experiment data showed that the highest EPA (1.221%) and DHA (0.312%) content were reached at 50 ℃ and 24 h with 150 rpm of agitation. However, through the RSM-Box Bhenken analysis and 3D surface plot, it wa...

Substrate and product concentration data obtained by simulating enzyme-substrate reaction rate equations were used to test two proposed kinetic rate constant estimation techniques in this study. In the first technique, the turnover number, k3, was calculated using early transient time domain data, which are difficult to obtain experimentally. The technique used an iterative approach to calculate k3 with a pair of data and the value of k3 could be retrieved with 35% error. The second technique calculated k3 using stationary domain data and the value of k3 could be retrieved with less than 5% error. This second technique also offered internal consistency in the calculation of k3 by calculating k3 both from the intercept and the slope of the linear plot derived in this study. A series of sensitivity analyses was conducted to understand the robustness of the second technique in estimating k3 from simulated data to the changes in the reaction rate constants (k1, k2, and k3) and the initi...

Particle design is presently a major development of supercritical fluids applications, mainly in the paint, cosmetic, pharmaceutical, and specialty chemical industries. Supercritical fluids have been successfully used to obtain composites or encapsulates, which comprise an active compound loaded into a matrix of a carrier material, in order to improving product preservation as well as controlling the dissolution rate of the active compound. The particles from gas-saturated solution (PGSS) process has been used to micronize suspensions of different substances in polymer melts, in order to obtain composite materials. The particle formation of functional pigment with biodegradable polymer, polyethelene glycol (PEG) was performed by supercritical carbon dioxide (SCO 2) in a thermostated stirred vessel. PGSS were carried out in different temperatures and pressures to measure the optimum condition for the formation of functional pigment particle (pressure from 20 MPa to 25 MPa, temperature from 40 to 50°C). The average diameter of the particles obtained at different conditions was about 0.8-1.4 µm accompanied by the size reduction, comparing the size of unprocessed particles, 430 µm.

Most of the health benefits of fish oil can be attributed to the presence of omega-3 fatty acids like Docosahexenoic acid (DHA) and Eicosapentaenoic acid (EPA). There are few dietary sources of EPA and DHA other than oily fish. EPA and DHA have great potential effect on human health. In this research, Supercritical carbon dioxide (scCO2) extracted mackerel oil was reacted by enzyme at different systems to improve the EPA and DHA. Different types of immobilize enzyme TL-IM, RM-IM, Novozyme 435 were assessed for improving PUFAs. Best result was found at non-pressurized system using TL-IM. Reacted oil particle were obtained with polyethylene glycol by gas saturated solution process (PGSS). Different parameters like temperature, pressure, agitation speed and nozzle size effect on particle formulation were observed. SEM and PSA analysis showed, small size non spherical particles were obtained. It was found that after particle formation poly unsaturated fatty acids (PUFAs) were present in particle as same in oil. PUFAs release from particle was almost linear against constant time duration. Oil quality in particle not change significantly, in this contrast this study will be helpful for food and pharmaceutical industry to provide high EPA and DHA containing powder.

MAGs with caprylic acid (CA) catalyzed by the 1,3 specific lipase DF, from Rhizopus oryzae, immobilized on Accurel MP1000, to produce STAGs with the structure CA-PUFA-CA, and (4) purification of STAGs by the neutralization of free fatty acids (FFAs) and extraction of STAGs with hexane.