de Moura Bell Juliana M L N, Aquino Leticia F M C, Liu Yan, Cohen Joshua L, Lee Hyeyoung, de Melo Silva Vitor L, Rodrigues Maria I, Barile Daniela
Department of Food Science and Technology, University of California, Davis, One Shields Avenue, Davis 95616; Foods for Health Institute, University of California, Davis, One Shields Avenue, Davis 95616.
Department of Food Science and Technology, Universidade Federal Fluminense, Niteroi, Rio de Janeiro, 24230340, Brazil.
J Dairy Sci. 2016 Aug;99(8):6157-6163. doi: 10.3168/jds.2016-11065. Epub 2016 May 26.
Enzymatic hydrolysis of lactose has been shown to improve the efficiency and selectivity of membrane-based separations toward the recovery of bioactive oligosaccharides. Achieving maximum lactose hydrolysis requires intrinsic process optimization for each specific substrate, but the effects of those processing conditions on the target oligosaccharides are not well understood. Response surface methodology was used to investigate the effects of pH (3.25-8.25), temperature (35-55°C), reaction time (6 to 58 min), and amount of enzyme (0.05-0.25%) on the efficiency of lactose hydrolysis by β-galactosidase and on the preservation of biologically important sialyloligosaccharides (3'-siallylactose, 6'-siallylactose, and 6'-sialyl-N-acetyllactosamine) naturally present in bovine colostrum whey permeate. A central composite rotatable design was used. In general, β-galactosidase activity was favored at pH values ranging from 3.25 to 5.75, with other operational parameters having a less pronounced effect. A pH of 4.5 allowed for the use of a shorter reaction time (19 min), lower temperature (40°C), and reduced amount of enzyme (0.1%), but complete hydrolysis at a higher pH (5.75) required greater values for these operational parameters. The total amount of sialyloligosaccharides was not significantly altered by the reaction parameters evaluated, suggesting specificity of β-galactosidase from Aspergillus oryzae toward lactose as well as the stability of the oligosaccharides at pH, temperature, and reaction time evaluated.
乳糖的酶促水解已被证明可提高基于膜分离的生物活性低聚糖回收效率和选择性。实现最大程度的乳糖水解需要针对每种特定底物进行内在工艺优化,但这些加工条件对目标低聚糖的影响尚未得到充分了解。采用响应面法研究了pH值(3.25 - 8.25)、温度(35 - 55°C)、反应时间(6至58分钟)和酶用量(0.05 - 0.25%)对β-半乳糖苷酶水解乳糖效率以及对牛初乳清渗透物中天然存在的具有生物学重要性的唾液酸低聚糖(3'-唾液酸乳糖、6'-唾液酸乳糖和6'-唾液酸-N-乙酰乳糖胺)保存情况的影响。使用了中心复合旋转设计。一般来说,β-半乳糖苷酶活性在pH值3.25至5.75范围内较高,其他操作参数的影响较小。pH值为4.5时,可以使用较短的反应时间(19分钟)、较低的温度(40°C)和较少的酶用量(0.1%),但在较高pH值(5.75)下完全水解需要这些操作参数取更大的值。所评估的反应参数对唾液酸低聚糖的总量没有显著影响,这表明米曲霉β-半乳糖苷酶对乳糖具有特异性,以及在所评估的pH值、温度和反应时间下低聚糖的稳定性。