Preparation of Cross-Linked Enzyme Aggregates of Trehalose Synthase via Co-aggregation with Polyethyleneimine
文献类型: 外文期刊
作者: Zheng, Jianfeng 1 ; Chen, Ying 1 ; Yang, Liwei 1 ; Li, Mingchun 2 ; Zhang, Jun 1 ;
作者机构: 1.Tianjin Acad Agr Sci, Tianjin Inst Forestry & Pomol, Tianjin 300384, Peoples R China
2.Nankai Univ, Coll Life Sci, Tianjin 300071, Peoples R China
关键词: Trehalose synthase;Polyethyleneimine;Co-aggregation;Cross-linking
期刊名称:APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY ( 影响因子:2.926; 五年影响因子:2.685 )
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收录情况: SCI
摘要: Trehalose synthase (TreS) from Meiothermus ruber was co-aggregated with polyethyleneimine (PEI) and precipitated with polyethylene glycol (PEG), followed by cross-linking with glutaraldehyde to obtain TreS-polyethyleneimine cross-linked enzyme aggregates (termed as CLEAs-PEI-PEG). The TreS solution at 0.5 mg mL~(?1) protein concentration, with PEI at a mass ratio of 1:0.8 (enzyme/PEI, w/w) and 25 % (w/v) PEG concentration were found to be most adequate for the co-aggregation of TreS. CLEAs-PEI-PEG was most active with glutaraldehyde at a mass ratio of 1:0.5 (enzyme/glutaraldehyde, w/w) to cross-link the co-aggregates. The CLEAs-PEI-PEG prepared in this work had an optimum pH of 6.5 and optimum temperature of 60 ℃. For lower concentrations of enzyme, using PEI could enhance the cross-linking efficiency of TreS. The thermal stability and pH tolerance of CLEAs-PEIPEG were significantly improved. Scanning electron microscopy revealed that the main structure of CLEAs-PEI-PEG showed scaffolding morphology which was constituted by structured ball-like particles with a size of 1–2.5 μm in diameter.
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