Conditions and optimization for the degradation of different substrates by a carotenoid cleavage enzyme
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摘要: 利用高效液相色谱研究类胡萝卜素裂解酶催化不同底物生成的非香气物质随酶解时间、p H、温度的变化规律,结果表明非香气物质生成量会随着酶解时间的增加、p H的降低和温度的升高出现先增加后减少的趋势。另外发现β-胡萝卜素在不同反应p H下可生成不同的裂解产物。利用二次多项式逐步回归得到酶解的最佳工艺条件,β-胡萝卜素、玉米黄质、β-阿朴-8’-胡萝卜醛、角黄质和虾青素的酶解条件分别为p H4.5、温度30℃、时间15 min,p H4.5、温度30℃、时间13 min,p H4.5、温度33℃、时间45 min,p H4.0、温度30℃、时间41 min,p H4.5、温度50℃、时间7 min。按上述条件进行酶解反应,得到产物β1、z1和z2、a1、c1和c2、x1和x2的生成量分别为定义条件下的1.74、1.80、1.39、1.66和1.75倍。该方法有效提高了非香气物质的生成量,并节约了时间。Abstract: The variation of the non- volatile compounds generated from the degradation of different substrates by carotenoid cleavage enzyme was investigated using HPLC method,with the changing of enzymolysis time,p H value and temperature.The results showed that the production of non- volatile compounds had a trend of increased at the beginning and then decreased,with the increasing of enzymolysis time and temperature and decreasing of p H.In addition,it was found that the β- carotene was degraded into different products at different p H values. By using quadratic polynomial stepwise regression analysis,the optimum reaction conditions for cleaving different substrates by purified enzyme were obtained and summarize as follows: p H4.5,15 min at 30 ℃ for β- carotene,p H4.5,13 min at 30 ℃ for zeaxanthin,p H4.5,45 min at 33 ℃ for β- apo- 8'- carotenal,p H4.0,41 min at 30 ℃ for canthaxanthin,p H4.5,7 min at 50 ℃ for astaxanthin,respectively. Under these conditions,the total yields of nonvolatile products were 1.74,1.80,1.39,1.66 and 1.75- fold higher than the defined yields,respectively.In conclusion,this method could be used to enhance the yields of non- volatile products effectively and was also time- saving.
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