Department of Physical and Chemical Sciences, Elizade University, Ilara-Mokin, Nigeria; Department of Biochemistry and Molecular Biology, Obafemi Awolowo University, Ile-Ife, Nigeria.
Biocatalysis and Technical Biology Research Group, Institute of Biomedical and Microbial Biotechnology, Cape Peninsula University of Technology, South Africa.
Int J Biol Macromol. 2019 Nov 1;140:833-841. doi: 10.1016/j.ijbiomac.2019.08.159. Epub 2019 Aug 21.
Industrial enzymes such as α-amylase must be thermostable and also easily purified/concentrated. Hence, aqueous two-phase partitioning systems (ATPS) was exploited for the partitioning of α-amylase from Aureobasidium pullulans due to its numerous advantages over conventional purification strategy. A. pullulans α-amylase was partially purified using ATPS via response surface methodology (RSM). The potentials of the ATPS-purified enzyme for possible industrial application such as resistance to thermal inactivation was investigated in comparison with the crude enzyme. PEG-6000 was the polymer of choice for ATPS as it resulted in higher purification factor (PF), %yield (Y), and partition coefficient (PC). At optimum levels (% w/v) of 20, 12 and 7.5 for PEG-6000, sodium citrate and sodium chloride respectively, maximum PF, Y and PC of 4.2, 88%, and 9.9 respectively were obtained. The response model validation and reliability were established based on the closeness between the experimented and predicted values. The kinetic and thermodynamic parameters such as Q, t, k, D - value, E, [Formula: see text] [Formula: see text] of the ATPS-purified α-amylase indicated that it was thermostable at 50 to 60 °C compared to the crude α-amylase. A thermodynamically stable and ATPS-purified α-amylase from A. pullulans has properties easily applicable for most industrial processes.
工业酶，如α-淀粉酶，必须具有热稳定性，并且易于纯化/浓缩。因此，由于其相对于传统纯化策略的众多优势，双水相分配系统（ATPS）被用于从出芽短梗霉中分配α-淀粉酶。使用 ATPS 通过响应面法（RSM）对出芽短梗霉α-淀粉酶进行部分纯化。通过与粗酶相比，研究了 ATPS 纯化酶在可能的工业应用（如耐热失活）方面的潜力。PEG-6000 是 ATPS 的首选聚合物，因为它导致更高的纯化因子（PF）、%产率（Y）和分配系数（PC）。在 PEG-6000、柠檬酸钠和氯化钠的最佳浓度（% w/v）分别为 20、12 和 7.5 时，获得了最大的 PF、Y 和 PC，分别为 4.2、88%和 9.9。基于实验值和预测值之间的接近程度，建立了响应模型验证和可靠性。ATPS 纯化的α-淀粉酶的动力学和热力学参数，如 Q、t、k、D 值、E、[公式：见正文] [公式：见正文]，表明它在 50 至 60°C 时比粗α-淀粉酶更耐热。与粗α-淀粉酶相比，从出芽短梗霉中获得的热力学稳定和 ATPS 纯化的α-淀粉酶具有易于适用于大多数工业过程的性质。
