In this paper, glutamate dehydrogenase (Gldh) is reported to efficiently display on Escherichia coli cell surface by using N-terminal region of ice the nucleation protein as an anchoring motif. The presence of Gldh was confirmed by SDS-PAGE and enzyme activity assay. Gldh was detected mainly in the outer membrane fraction, suggesting that the Gldh was displayed on the bacterial cell surface. The optimal temperature and pH for the bacteria cell-surface displayed Gldh (bacteria-Gldh) were 70 degrees C and 9.0, respectively. Additionally, the fusion protein retained almost 100% of its initial enzymatic activity after 1 month incubation at 4 degrees C. Transition metal ions could inhibit the enzyme activity to different extents, while common anions had little adverse effect on enzyme activity. Importantly, the displayed Gldh is most specific to L-glutamate reported so far. The bacterial Gldh was enabled to catalyze oxidization of L-glutamate with NADP(+) as cofactor, and the resultant NADPH can be detected spectrometrically at 340 nm. The bacterial-Gldh based L-glutamate assay was established, where the absorbance at 340 nm increased linearly with the increasing L-glutamate concentration within the range of 10-400 mu M. Further, the proposed approach was successfully applied to measure L-glutamate in real samples. (C) 2014 Elsevier Inc. All rights reserved.