A rapid preparative method for the purification of vitamin E acetate by kinetic reverse phase chromatography

Alpha-tocopherol or vitamin E is an essential food ingredient. Due to high purity requirement in food applications, it is majorly produced in the acetate form through chemical routes. However, the isolation of food grade vitamin E acetate (VEA) is critical due to the co-generation of close characteristic impurities in the production processes. The work undertaken here provides a new kinetic chromatography method for production of food grade VEA. In this work, suitable adsorbent for chromatography separation was selected based on shallow-bed binding–elution characteristics. Loading conditions were identified by frontal adsorption behavior. Kinetic parameters and bed height for the purification was predicted by establishing the relationship between bed height, purity and the relative separation factor (RSF). The RSF of 2.8 and corresponding bed height is predicted from the established relationship for the separation of VEA from impurities. The bed height of 170 cm with isocratic mobile phase of 2% (wt/wt) water in methanol provides 98 wt.% purity of VEA with recovery of 90%. This work has highlighted methodology based on kinetic parameters for the separation of close characteristic impurities from target molecule. This work also provided the application of simple handy tools like RSF and dual wavelength detection for the rapid development of preparative separation process for VEA and other such valuable components from unknown impurities

Green synthesis of isopropyl myristate in novel single phase medium Part I: Batch optimization studies

Isopropyl myristate finds many applications in food, cosmetic and pharmaceutical industries as an emollient, thickening agent, or lubricant. Using a homogeneous reaction phase, non-specific lipase derived from Candida antartica, marketed as Novozym 435, was determined to be most suitable for the enzymatic synthesis of isopropyl myristate. The high molar ratio of alcohol to acid creates novel single phase medium which overcomes mass transfer effects and facilitates downstream processing. The effect of various reaction parameters was optimized to obtain a high yield of isopropyl myristate. Effect of temperature, agitation speed, organic solvent, biocatalyst loading and batch operational stability of the enzyme was systematically studied. The conversion of 87.65% was obtained when the molar ratio of isopropyl alcohol to myristic acid (15:1) was used with 4% (w/w) catalyst loading and agitation speed of 150 rpm at 60 °C. The enzyme has also shown good batch operational stability under optimized conditions

Green synthesis of isopropyl myristate in novel single phase medium Part II: Packed bed reactor (PBR) studies

Isopropyl myristate is a useful functional molecule responding to the requirements of numerous fields of application in cosmetic, pharmaceutical and food industry. In the present work, lipase-catalyzed production of isopropyl myristate by esterification of myristic acid with isopropyl alcohol (molar ratio of 1:15) in the homogenous reaction medium was performed on a bench-scale packed bed reactors, in order to obtain suitable reaction performance data for upscaling. An immobilized lipase B from Candida antartica was used as the biocatalyst based on our previous study. The process intensification resulted in a clean and green synthesis process comprising a series of packed bed reactors of immobilized enzyme and water dehydrant. In addition, use of the single phase reaction system facilitates efficient recovery of the product with no effluent generated and recyclability of unreacted substrates. The single phase reaction system coupled with a continuous operating bioreactor ensures a stable operational life for the enzyme

Dataset of gene cloning and gel filtration chromatography of R-est6

The data presented in this article are connected to the research article entitled “Expression, purification and biochemical characterization of a family 6 carboxylesterase from Methylococcus capsulatus (bath)” (Soni et al., 2016 [1]). The family 6 carboxylesterases are the smallest and display broad substrate specificity. The 1 kb gene encoding, a family 6 carboxylesterase – R-est6, was amplified from the genome of M. capsulatus (bath strain), and showed in the agarose gel. The corresponding purified protein, after overexpression in Escherichia coli, was biochemically studied in the research article (Soni et al., 2016 [1]). R-est6 has hydrophobic patches on the surface so, it is expected to show oligimeric forms. Here, we have confirmed the presence of oligomers by gel filtration chromatography data and the proteins belonging to the different peaks are shown on a SDS-PAGE

Synthesis of designer triglycerides by enzymatic acidolysis

Enzymatic acidolysis process was developed for modification of fully hydrogenated soybean oil (FHSO) by incorporation of caprylic acid, a medium chain fatty acid. Immobilized sn-1,3 specific lipase PyLip was used to modify FHSO to produce a new fat with improved physico-chemical and functional properties. PyLip mediated acidolysis resulted in 88% reduction of substrate triglycerides and 45.16% incorporation of caprylic acid in FHSO at molar ratio of 1:3 of FHSO and caprylic acid in 60 min reaction time. HPLC analysis revealed formation of mono-substituted and di-substituted TAGs post enzymatic acidolysis. Physical properties of synthesized lipid were studied using DSC and XRD and considerable change was observed in the final product compared to the starting material. The present study reports a faster acidolysis process in the presence of solvent enhancing the modification of FHSO with caprylic acid and having no side products formation (monoglycerides and diglycerides) making the entire process highly efficient and commercially attaractive

An integrated process for the extraction of fuel and chemicals from marine macroalgal biomass

We describe an integrated process that can be applied to biomass of the green seaweed, Ulva fasciata, to allow the sequential recovery of four economically important fractions; mineral rich liquid extract (MRLE), lipid, ulvan, and cellulose. The main benefits of our process are: a) its simplicity and b) the consistent yields obtained from the residual biomass after each successive extraction step. For example, dry Ulva biomass yields ~26% of its starting mass as MRLE, ~3% as lipid, ~25% as ulvan, and ~11% as cellulose, with the enzymatic hydrolysis and fermentation of the final cellulose fraction under optimized conditions producing ethanol at a competitive 0.45 g/g reducing sugar. These yields are comparable to those obtained by direct processing of the individual components from primary biomass. We propose that this integration of ethanol production and chemical feedstock recovery from macroalgal biomass could substantially enhance the sustainability of marine biomass use