Effective immobilisation of lipase to enhance esterification potential and reusability
-
Ashok Kumar
,
Vikrant Sharma
Abstract
A commercial lipase, “Lipolase T100”, was immobilised onto silica by means of physical adsorption. The silica-bound lipase was subsequently exposed to 1 vol. % glutaraldehyde (pentane-1,5-dial). The silica was loaded repeatedly with the Lipolase T100 in 0.05 M Tris buffer (pH 8.5) until saturation was achieved. During the 1st, 2nd, 3rd, 4th, and 5th cycles of loading of silica with the enzyme, the protein-binding on the silica achieved 51.73 %, 48.27 %, 26.92 %, 10.73 %, and 4.29 %, respectively. The synthesis of methyl salicylate (methyl 2-hydroxybenzoate) and linalyl ferulate (3,7-dimethylocta-1,6-dien-3-yl 4-hydroxy-3-methoxycinnamate) carried out at 45°C under shaking with mole ratios of 200 mM of acid and 500 mM alcohol in DMSO using 15 mg mL−1 of hyper-activated biocatalyst resulted in yield(s) of 77.2 % of methyl salicylate and 65.3 % of linalyl ferulate in the presence of molecular sieves. The hyper-activated biocatalyst was more efficient than the previously reported silica-bound lipase with minimum leaching of the enzyme from the reaction mixture. The K m and V max of the free (0.142 mM and 38.31 μmol min−1 mL−1, respectively) and silica-bound lipase (0.043 mM and 26.32 μmol min−1 mg−1, respectively) were determined for the hydrolysis of p-NPP. During repeated esterification studies using silica-bound lipase, yields of 50.1 % of methyl salicylate after the 5th cycle, and 53.9 % of linalyl ferulate after the 7th cycle of esterification were recorded. In the presence of molecular sieves (30 mg mL−1) in the reaction mixture, the maximum syntheses of methyl salicylate (77.2 %) and linalyl ferulate (65.3 %) were also observed. In a volumetric batch scale-up, when the reaction volume was increased to 50 mL, 44.9 % and 31.4 % yields of methyl salicylate and linalyl ferulate, respectively, were achieved.
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© 2013 Institute of Chemistry, Slovak Academy of Sciences
Articles in the same Issue
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Articles in the same Issue
- Flexibility of active-site gorge aromatic residues and non-gorge aromatic residues in acetylcholinesterase
- Influence of trace elements supplementation on the production of recombinant frutalin by Pichia pastoris KM71H in fed-batch process
- Mesoporous nanocrystalline MgAl2O4: A new heterogeneous catalyst for the synthesis of 2,4,6-triarylpyridines under solvent-free conditions
- Effective immobilisation of lipase to enhance esterification potential and reusability
- Hydrogen production by steam reforming of glycerol over Ni/Ce/Cu hydroxyapatite-supported catalysts
- Solvent-free acetylation and tetrahydropyranylation of alcohols catalyzed by recyclable sulfonated ordered nanostructured carbon
- Pertraction of methylene blue using a mixture of D2EHPA/M2EHPA and sesame oil as a liquid membrane
- Selective separation of essential phenolic compounds from olive oil mill wastewater using a bulk liquid membrane
- Evaluation of temperature effect on growth rate of Lactobacillus rhamnosus GG in milk using secondary models
- Fastener effect on magnetic properties of chain compounds of dinuclear ruthenium carboxylates
- Synthesis of novel fluorene-functionalised nanoporous silica and its luminescence behaviour in acidic media
- d-Glucosamine as an efficient and green additive for palladium-catalyzed Heck reaction
- Anti-oxidative properties of bi-1,2,4-triazine bisulphides
