Fixation and stabilization of Escherichia coli cells displaying genetically engineered cell surface proteins.
Fixation and stabilization of Escherichia coli cells displaying genetically engineered cell surface proteins.
复制标题
展示基因工程细胞表面蛋白的大肠杆菌细胞的固定和稳定。
DOI：
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发表时间：
2000
影响因子：
3.8
通讯作者：
G. Georgiou
中科院分区：
文献类型：
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作者：
A. Freeman;S. Abramov;G. Georgiou
A large biotechnological potential is inherent in the display of proteins (e.g., enzymes, single-chain antibodies, on the surface of bacterial cells) (Georgiou et al., 1993). Applications such as immobilized whole-cell biocatalysts or cellular adsorbents require cell fixation to prevent disintegration, stabilization of the anchored protein from leakage, denaturation or proteolysis, and total loss of cell viability, preventing medium and potential product contamination with cells. In this article we describe the adaptation of a simple two-stage chemical crosslinking procedure based on "bi-layer encagement" (Tor et al., 1989) for stabilizing Escherichia coli cells expressing an Lpp-OmpA (46-159)-beta-lactamase fusion that displays beta-lactamase on the cell surface. Bilayer crosslinking and coating the bacteria with a polymeric matrix is accomplished by treating the cells first with either glutaraldehyde or polyglutaraldehyde, followed by secondary crosslinking with polyacrylamide hydrazide. These treatments resulted in a 5- to 25-fold reduction of the thermal inactivation rate constant at 55 degrees C of surface anchored beta-lactamase and completely prevented the deterioration of the cells for at least a week of storage at 4 degrees C. The stabilization procedure developed paves the way to scalable biotechnological applications of E. coli displaying surface anchored proteins as whole-cell biocatalysts and adsorbents.
一种快速、灵敏的细胞酶联免疫吸附测定 (CELISA)，用于检测和定量针对细胞表面决定簇的抗体。
影响因子：
2.2
作者：
Walker,KW;Llull,R;Balkian,GK;Ko,HS;Flores,KM;Ramsamooj,R;Black,KS;Hewitt,CW;Martin,DC
通讯作者：
Martin,DC