Carla Cristina de Sousa, Amanda Carmelo da Rocha, Larissa Nayhara Soares Santana Falleiros, Eloízio Júlio Ribeiro, Miriam Maria de Resende
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引用次数: 0
摘要
半乳糖寡糖(GOS)是不可消化的,具有益生元作用。它们刺激有益细菌的活动,有助于肠道健康。它们可以通过酶合成,β-半乳糖苷酶是进行半乳糖基化的酶。游离形式的酶会干扰这一过程,因此采用固定化方法。共价键是其中一种方法,二氧化硅作为一种载体得到了突出的应用。因此,利用商业β-半乳糖苷酶以其自由形态产生并在二氧化硅上固定化,对GOS的合成进行了评价。评估缓冲离子强度和乳糖浓度。采用析因设计评价初始乳糖浓度、搅拌和温度对GOS合成的影响。考察了磁搅拌、机械搅拌和固定床反应器构型对GOS合成的影响。在高浓度乳糖(400 g L-1)条件下,游离酶反应时间最短(3 h),固定化酶反应时间最长(6 h)。分数析因设计变量的影响均为正,其中搅拌影响最大,GOS浓度最高为67.70 g L-1。固定化酶在磁力搅拌和机械搅拌条件下的合成浓度分别为70.81 g L-1和90.37 g L-1。在固定床上的实验中,GOS的浓度保持在30 g L-1以上,表明在二氧化硅上固定化β-半乳糖苷酶是很有希望合成GOS的方法。
New insights for the synthesis of galacto-oligosacharides with β-Galactosidase immobilized in silica.
Galacto-oligosaccharides (GOS) are non-digestible and have a prebiotic effect. They stimulate the activity of beneficial bacteria and contribute to intestinal health. They can be synthesized enzymatically, with β-galactosidase being the enzyme that performs transgalactosylation. Enzymes in their free form can interfere with the process, so immobilization methods are used. Covalent bonding is one of these methods and silica has gained prominence as a carrier. Therefore, the synthesis of GOS was evaluated using commercial β-galactosidase produced in its free form and immobilized on silica. Buffer ionic strength and lactose concentrations were assessed. A Factorial Design was proposed to evaluate the influence of initial lactose concentration, agitation, and temperature on the synthesis of GOS. Magnetic and mechanical stirring and fixed-bed reactor configuration were also evaluated in the synthesis of GOS. GOS were produced in high concentrations of lactose (400 g L-1), in the shortest time (3 h) for free enzyme and longest for immobilized enzyme (6 h). The effects of the Fractional Factorial Design variables were positive, the most effective influence was stirring, and the highest concentration of GOS was 67.70 g L-1. GOS was synthesized with the immobilized enzyme at a concentration of 70.81 g L-1 when using magnetic stirring and 90.37 g L-1 with mechanical stirring. GOS concentrations remained at values higher than 30 g L-1 in experiments in a fixed bed showing that β-Galactosidase immobilized on silica is promising for synthesizing GOS.
期刊介绍:
Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects.
Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence.
Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".