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Sequence analysis of Candida rugosa lipase 1 (LIP1) predicts the presence of three N-linked glycosylation sites at asparagine 291, 314, 351. To investigate the relevance of sugar chains in the activation and stabilization of LIP1, we directed site mutagenesis to replace the above mentioned asparagine with glutamine residues. Comparison of the activity of mutants with that of the wild-type (wt) lip
Rhizopus arrhizus lipase immobilized on porous polypropylene was used in an acidolysis reaction in toluene at low water activity (0.11) to exchange the fatty acid in the sn-1-position of digalactosyldiacylglycerol (DGDG). Without extra precautions, an yield of only about 20% was obtained, mainly due to side reactions; incorporation of an extra acyl group on the primary hydroxyl of the digalactosyl
The enzymatic syntheses of 1-lauroyl-dihydroxyacetone and 1,3-dilauroyl-dihydroxyacetone were investigated. Lipase B from Candida antarctica (SP435) was used to catalyse the acylation of dihydroxyacetone (DHA) with lauric acid in organic solvent media at controlled water activity. High conversions of dihydroxyacetone (> 90%) are achieved when the water activity is 0.11 or below in solvents of vari
Immobilized lipase from Candida antarctica lipase B (Novozym 435) was effective in the synthesis of lysophosphatidylcholine (LPC). The transesterification of L-α-glycerophosphorylcholine (GPC) and vinyl laurate was carried out in a solvent free system or in the presence of 50% (v/v) t- butanol. High conversions (>95%) were easily achieved. The lipase was selective for the sn-1 position of the glyc
Six different lipases were screened for their ability of acidolysis between digalactosyldiacylglycerol (DGDG) and heptadecanoic acid in toluene. Lipases from Geotrichum candidum, Alcaligenes sp. and Penicillium camembertii did not catalyse the acidolysis reaction. Rhizopus arrhizus and Rhizomucor miehei (Lipozyme) catalysed the acidolysis but produced a mixture of DGMG, DGDG, acyl-DGMG and acyl-DG
A combination of two enzymes, phospholipase D (PL D) and C (PL C), was investigated for the production of two lysophospholipids, 1-lauroyl-rac- glycerophosphate (1-LGP) and 1-lauroyl-dihydroxyacetonephosphate (1-LDHAP). The high transphosphatidylation ability of phospholipase D from Streptomyces sp. allowed the formation of 1-lauroyl-phosphatidylglycerol (1-LPG) and 1- lauroyl-phosphatidyldihydrox
Two different immobilisation techniques for lipases were investigated: adsorption on to Accurel EP-100 and deposition on to Celite. The specific activities were in the same order of magnitude, 2.9 (μmol min -1 mg protein) when Celite was used as support and 2.3 (μmol min -1 mg -1 protein) when Accurel EP-100 was used as support,
Enzymatic synthesis of cetyl palmitate was performed in a solvent-free system at 65 °C using immobilized Candida antarctica lipase. Batch reactions at controlled water activity showed that the yield could be increased from 88.8 to 99.1% by decreasing the water activity from 1 to 0.05. A continuous reactor configuration was constructed, where two tubular reactors were run in sequence with a separat
(S)-Hydroxynitrile lyases from Hevea brasiliensis, Manihot esculenta, and Sorghum bicolor and (R)-hydroxynitrile lyase from Prunus amygdalus have been used as biocatalyst for the enantiospecific addition of hydrogen cyanide to aldehydes in organic solvents. The effects of the reaction parameters on the enzymatic reaction rate and product enantiomeric excess (e.e.) are presented. The reaction rate
Immobilised 1,3-specific lipase from Rhizopus arrhizus was used as catalyst for the esterification of dl-glycero-3-phosphate and fatty acid or fatty acid vinyl ester in a solvent-free system. With lauric acid vinyl ester as acyl donor, a(w)<0.53 favored the synthesis of lysophosphatidic acid (1-acyl-rac-glycero-3-phosphate, LPA1) and the spontaneous acyl migration of the fatty acid on the molecule
α-Chymotrypsin immobilized on celite catalyzing a kinetically controlled dipeptide synthesis reaction in acetonitrile medium showed an odd behavior in response to additions of triethylamine to the reaction mixture. This base is used to deprotonate the nucleophilic reagent, l-alaninamide hydrochloride, in order to increase its nucleophilicity and solubility. However, the enzyme performance is appar
The influence of different reaction systems on α-chymotrypsin-catalyzed synthesis of eledoisin and LH-RH peptides from (7+4) and (5+5) fragments was investigated. The peptide yield was determined in the following systems: buffered aqueous media, frozen solutions, organic media, and cosolvent mixtures. The experimental set up was tailored to allow the screening of an array of conditions with minimu
The stereoselective reduction of 2-pentanone by alcohol dehydrogenase from Thermoanaerobium brockii was studied at controlled water activity and at different reaction temperatures. The reaction rate increased when water activity was increased from 0.32 to 0.96 and when raising the temperature from 5°C to 40°C. The enantioselectivity, E, reached a plateau value at high water activities. The enantio
Hydroxy carboxylate viologen oxidoreductase (HVOR) contained in Proteus vulgaris cells was used in the reduction of chloropyruvic acid to D-(S)-chlorolactic acid. A synthetic electron mediator 1,1'-carbamoylmethylviologen (CAV) was used to transfer electrons to HVOR. Three different methods for regenerating the mediator were evaluated: (1) electrochemically driven reduc
We have identified and characterized the temporal and spatial regulation of a plasma membrane arabinogalactan protein epitope during development of the aerial parts of oilseed rape using the monoclonal antibody JIM8. The JIM8 epitope is expressed by the first cells of the embryo and by certain cells in the sexual organs of flowers. During embryogenesis, the JIM8 epitope ceases to be expressed by t
Mass transfer limitations were studied in enzyme preparations of α- chymotrypsin made by deposition on different porous support materials such as controlled pore glasses, Celite, and polyamides of different particle sizes. It is the onset of mass transfer limitations that determines the position of the activity optimum with respect to enzyme loading on each support. The evidence of various experim
