Synthesis And Analysis Carnosine And The Simulacrum By Enzyme In Micro-aqueous Phase

It has a promising profit of synthesis by enzyme in micro-aqueous phase, for the more mild operational condition, good selectivity, lower cost, solvent and catalyzer recycling, high optical purity of production and etc. The carnosine (β-Ala-L-His) has antioxidant activity using wildly in biology, chemical engineer, drug and etc. They are a kind of foreground of developing biological composition, and the researching and developing hotspot all over the world.In this paper we study the synthesis of the carnosine and the simulacrum using n-Butanol or THF as organic impregnant,α-Chymotrypsin as catalyzer, L-His or 1H-imidazole-4,5-dicarboxylic acid and L-Ala as substrate. Analysis and confirm the spectrum and TLC. Review the conditions of carnosine and the simulacrum by the numbers, the optimum conditions of the simulacrum are obtained from the orthogonal test.The result showed thatα-Chymotrypsin can respectively catalyze carnosine and the simulacrum in the micro-aqueous by using n-Butanol and THF.There is a new component in the sample by TLC. The UV spectra of the component absorb at 240, 310 and 330nm. The reserve time is 4.25min detected by HPLC. There are five groups displacement in 1H NMR spectra. We can confirm that the component is 5-(1-Carboxy-ethylcarbamoyl)- 4-carboxylic acid -imidazole.The result of single factor tests of the simulacrum showed that the temperature is 35℃, the yield is 20.0%; the time is 70min, the yield is 46.4%. The light is not effect the yield.α-Chymotrypsin can keep activity in pH during 7.5 and 9.5, the yield is during 44.5% and 53.5%, at pH8.5 the yield is 55%. The dosage ofα-Chymotrypsin is 4% (weight proportion), the proportionment of 1H-imidazole-4,5-dicarboxylic acid and L-Ala is 1:1(weight proportion), the yield is 44.5%; the water contented in micro-aqueous phase is 10%, the yield is 42.3%; the proportion of 1H-imidazole-4,5-dicarboxylic to L-Ala is 1.5 to 1, the yield is 45.4%.The results of L25(65) orthogonal test showed that pH, temperature, the water contented in micro-aqueous phase, time, the proportion of materials are effected yield of synthesis the simulacrum markedly, the pH and temperature are effected the yield very markedly. For synthesis the simulacrum of carnosine, the optimum conditions are 35℃, 1.5h, the dosage of the enzyme is 1%, the water contented in micro-aqueous phase is...