Northeast Biotechnology Network/RENORBIO, Federal Rural University of Pernambuco (UFRPE), Dom Manoel de Medeiros, Dois Irmãos, 52171-900 Recife, PE, Brazil.
Academic Unit of Garanhuns/UAG, Federal Rural University of Pernambuco (UFRPE), Garanhuns, Av. Bom Pastor, Boa Vista, 55296-901 Garanhuns, PE, Brazil.
Int J Biol Macromol. 2020 May 1;150:922-929. doi: 10.1016/j.ijbiomac.2020.02.152. Epub 2020 Feb 15.
Pectinex Ultra SP-L, a commercial enzyme preparation with fructosyltransferase activity, was successfully immobilized by covalent binding to FeO-chitosan- magnetic nanoparticles. Immobilization carried out according to a 2-full factorial design where glutaraldehyde concentration, activation time and time of contact between enzyme and support were selected as the independent variables and immobilization yield as the response. The highest immobilization yield (94.84%) was obtained using 3.0% (v/v) glutaraldehyde and activation and contact times of 180 and 30 min, respectively. The immobilized biocatalyst, which showed for both hydrolytic and transfructosylating activities optimum pH and temperature of 7.0 and 60 °C, respectively, retained 70 and 86% of them after 6 cycles of reuse. A kinetic/thermodynamic study focused on thermal inactivation of the immobilized construct indicated high thermostability at temperatures commonly used for fructo-oligosaccharides (FOS) production. Maximum FOS concentration obtained in lab-scale experiments was 101.56 g L, with predominant presence of 1-kestose in the reaction mixture. The results obtained in this study suggest that the immobilized-enzyme preparation may be effectively exploited for FOS production and easily recovered from the reaction mixture by action of a magnetic field.
Pectinex Ultra SP-L 是一种具有果糖基转移酶活性的商业酶制剂,成功地通过共价键结合到 FeO-壳聚糖-磁性纳米粒子上进行固定化。固定化是根据 2 全因子设计进行的,其中戊二醛浓度、活化时间和酶与载体之间的接触时间被选为自变量,固定化产率为响应。使用 3.0%(v/v)戊二醛和分别为 180 和 30 min 的活化和接触时间,获得了最高的固定化产率(94.84%)。固定化生物催化剂的水解和转果糖基活性的最佳 pH 和温度分别为 7.0 和 60°C,重复使用 6 次后,其保留率分别为 70%和 86%。针对固定化结构的热失活动力学/热力学研究表明,在通常用于生产果寡糖(FOS)的温度下具有很高的热稳定性。在实验室规模的实验中获得的最大 FOS 浓度为 101.56 g/L,反应混合物中主要存在 1-蔗果三糖。本研究的结果表明,固定化酶制剂可有效地用于 FOS 的生产,并可通过磁场从反应混合物中轻松回收。