Improved phosphate biosorption by bacterial surface display of phosphate-binding protein utilizing ice nucleation protein
Improved phosphate biosorption by bacterial surface display of phosphate-binding protein utilizing ice nucleation protein
复制标题
利用冰核蛋白通过细菌表面展示磷酸盐结合蛋白来改善磷酸盐生物吸附
DOI：
10.1111/j.1574-6968.2009.01724.x
复制
发表时间：
2009-10-01
影响因子：
2.1
通讯作者：
Li, Lin
中科院分区：
文献类型：
--
作者：
Li, Qianqian;Yu, Ziniu;Li, Lin
传统的强化生物除磷（EBPR）系统往往在低化学需氧量（COD）或曝气条件下发生劣化。利用一种新鉴定的冰核蛋白的N端锚定结构（InaQ-N），将胞内磷酸盐结合蛋白（PBP）固定在恶臭假单胞菌或大肠杆菌的表面，从而提出了一种新的磷酸盐富营养化废水修复方法。表达了一种绿色荧光蛋白融合蛋白，并用于确认表面定位。然后将PBP靶向于E. coliJM 109和恶臭假单胞菌AB 92019。工程菌P. putida和E.大肠杆菌微生物能够以60和80 mg L-1的速率在5 h内对总磷酸盐进行绝对生物吸附。在重组恶臭假单胞菌细胞中，具有三个串联重复的InaQ-Ns的表面固定化PBP融合体表现出磷酸盐生物吸附的最大增量，与对照菌株相比为6倍。即使是热灭活的恶臭假单胞菌或大肠杆菌重组细胞。大肠杆菌保留了大量的生物吸附活性。目前的研究表明，细菌表面展示的PBP应被认为是一个强有力的竞争者产生的有机体能够在EBPR系统低COD下发挥作用，从而提高去除废水中的富营养化磷。
The conventional enhanced biological phosphorus removal (EBPR) system often deteriorates at low chemical oxygen demand ( COD) or under aeration conditions. A new approach that incorporates phosphate-eutrophic wastewater remediation was introduced through immobilization of an intracellular phosphate-binding protein (PBP) onto the surface of Pseudomonas putida or Escherichia coli, using the N-terminal anchor (InaQ-N) of a newly identified ice nucleation protein from Pseudomonas syringae. A green fluorescent protein-fusion protein was expressed and used to confirm surface localization. The PBP was then targeted to the surface of E. coli JM109 and P. putida AB92019. The engineered P. putida and E. coli microorganisms were capable of absolute biosorption of total phosphates at rates of 60 and 80 mg L-1, respectively, over 5 h. In the recombinant P. putida cells, a surface-immobilized PBP fusion that had three tandemly repeated InaQ-Ns exhibited the maximum increment in phosphate biosorption, at sixfold compared with the control strain. Even heat-killed recombinant cells of either P. putida or E. coli retained substantial biosorptive activities. The current study demonstrates that the bacterial surface display of PBP should be considered as a strong contender for generating organisms capable of functioning in EBPR systems under low COD, resulting in improved removal of eutrophic phosphorus from wastewaters.