Enzymatic synthesizing method of N-acetyl glycerol glutamate
A technology of glyceryl glutamate and synthesis method, applied in the field of enzymatic synthesis of glyceryl N-acetylglutamate, to achieve the effects of high biocompatibility, easy large-scale production, and economical raw materials
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Embodiment 1
[0021] Place 0.9458g (5mmol) N-acyl amino acid in a sandwich beaker, add 9.2g (100mmol) glycerol, heat up to 50°C, and add 0.2029g esterase (immobilized lipase Novozyme 435, immobilized lipase Lipozyme RM IM, immobilized lipase Lipozyme TL IM, papain, acyltransferase (Acylase) or trypsin), and reacted for 24 hours. After the reaction, add 5% NaCl solution 30ml and 0.5mol / 40ml of L sodium bicarbonate solution, stirred for 20-30min until no bubbles were generated, filtered, the filtrate was cooled to room temperature (25°C) and extracted with ethyl acetate, the organic phase was collected and dried to constant weight to obtain the N-acetyl Glutamic acid glycerides, the product is suitably diluted with mobile phase, adopts Waters 1525 high-performance liquid chromatographic analysis, the N-acyl amino acid amount of determination esterification, and calculates the esterification rate, the result is shown in Table 1; Chromatographic column is Lichrosher 100 RP -18 (5 μm), the colu...
Embodiment 2
[0027] 0.9458g N-acylglutamic acid, 9.2g glycerol, 0.2029g immobilized lipase Novozyme435, at different temperatures (30°C, 40°C, 50°C, 60°C, 70°C), the reaction time is 24 hours, other experimental operations are the same Example 1, the results are shown in Table 2.
[0028] Table 2 Effect of temperature on the activity of Novo 435 catalyzing the esterification of glutamic acid and glycerol
[0029]
[0030] Table 2 shows the 24-hour yields of N-acyl glutamic acid and glycerol esterification reaction catalyzed by immobilized lipase Novozyme 435 at different temperatures. Obviously, the effect of temperature on the rate of esterification is very significant, and the rate of esterification increases more than three times when the temperature increases from 30°C to 70°C. On the one hand, the catalytic activity of the enzyme is improved at high temperature, on the other hand, the solubility of N-acylglutamic acid in glycerol increases with the increase of temperature, and the...
Embodiment 3
[0032]0.9458g N-acylglutamic acid, 9.2g glycerol, the immobilized lipase Novozyme 435 of different addition amount (mass consumption is 0.5, 1%, 2%, 3%, N-acylglutamic acid and glycerol total mass meter, 5%, 10%), temperature 70 ℃, reaction time 24 hours, other operations are the same as embodiment 1, and the results are shown in Table 3.
[0033] The impact of table 3 enzyme dosage on the rate of esterification
[0034]
[0035] The results of the enzyme addition test show that the content of the final product does not increase significantly within the range of 0.5-3% enzyme dosage. Even if the enzyme dosage increases to 10%, the esterification rate only increases from 50% to 65%. This may be that the amount of enzymes has been saturated relative to the content of amino acids, so the effect of continuing to increase the amount of enzymes is not obvious. In addition, the reaction system has no solvent and the amount of glycerin is large, so the viscosity is relatively high...
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