Ramírez Matheus Alejandra O, Rivas Nilo
Instituto de Química y Tecnología, Facultad de Agronomía, Universidad Central de Venezuela.
Arch Latinoam Nutr. 2003 Jun;53(2):194-201.
The purpose of this work was to optimize the beta-galactosidase production by Kluyveromyces lactis, applying the Surface Response Methodology (SRM) and using deproteinized whey as fermentation medium. An Orthogonal Central Compound Design (OCCD) was used without repetition, with four factors: temperature, pH, agitation speed and fermentation time. Then, enzyme activity (U/ml) as response variable was used. Thirty trials in twenty-five treatments, with six repetitions at the central point, were carried out, in a New Brunswick Bioflo 2000 fermentor with a volume of 2 liters. The deproteinized whey obtained by thermocoagulation was chemically analyzed. The results were: moisture 93.83%, total solids 6.17%, protein 0.44%, lactose 4.85%, acidity 0.43% and pH 4.58. The best conditions in the enzyme production were: temperature 30.3 degrees C, pH 4.68, agitation speed 191 r.p.m. and fermentation time 18.5 h. with an enzyme production of 8.3 U/ml. The degree of purification obtained was 7.4 times and the yield was 50.8%. The purified enzyme had an optimum temperature of 60 degrees C and a pH of 6.2. This work shows that the yeast Kluyveromyces lactis grown in deproteinized whey is able to produce the enzyme beta-galactosidase and SRM can be used in the fermentology processes, specifically in determining the best suitable operation conditions.
本研究旨在通过应用表面响应法(SRM)并使用脱蛋白乳清作为发酵培养基,优化乳酸克鲁维酵母生产β-半乳糖苷酶的工艺。采用无重复的正交中心复合设计(OCCD),考察温度、pH、搅拌速度和发酵时间四个因素。然后,以酶活性(U/ml)作为响应变量。在一台容积为2升的新不伦瑞克Bioflo 2000发酵罐中进行了25种处理共30次试验,中心点重复6次。对通过热凝固法得到的脱蛋白乳清进行了化学分析。结果如下:水分93.83%,总固体6.17%,蛋白质0.44%,乳糖4.85%,酸度0.43%,pH 4.58。酶生产的最佳条件为:温度30.3℃,pH 4.68,搅拌速度191转/分钟,发酵时间18.5小时,酶产量为8.3 U/ml。纯化倍数为7.4倍,产率为50.8%。纯化后的酶最适温度为60℃,最适pH为6.2。本研究表明,在脱蛋白乳清中生长的乳酸克鲁维酵母能够产生β-半乳糖苷酶,并且表面响应法可用于发酵工艺,特别是用于确定最合适的操作条件。