Construction of a cell-surface display system based on the N-terminal domain of ice nucleation protein and its application in identification of mycoplasma adhesion proteins

2015 ◽  
Vol 119 (1) ◽  
pp. 236-244 ◽  
Author(s):  
S. Bao ◽  
S. Yu ◽  
X. Guo ◽  
F. Zhang ◽  
Y. Sun ◽  
...  
2022 ◽  
Vol 61 ◽  
pp. 102570
Author(s):  
João Vitor Dutra Molino ◽  
Roberta Carpine ◽  
Karl Gademann ◽  
Stephen Mayfield ◽  
Simon Sieber

2021 ◽  
Author(s):  
Qianqian Li ◽  
Tuantuan Wang ◽  
Yangzhi Ye ◽  
Shimin Guan ◽  
Baoguo Cai ◽  
...  

Abstract Objective To establish a temperature-induced chitosanase bacterial cell surface display system to produce chitooligosaccharides (COSs) efficiently for industrial applications. Results Temperature-inducible chitosanase CSN46A bacterial surface display systems containing one or two copies of ice nucleation protein (InaQ-N) as anchoring motifs were successfully constructed on the basis of Escherichia coli and named as InaQ-N-CSN46A and 2InaQ-N-CSN46A. The specific enzyme activity of 2InaQ-N-CSN46A reached 886.33±0.81 U/g cell dry weight, which was 45.6% higher than that of InaQ-N-CSN46A. However, few proteins were detected in 2InaQ-N-CSN46A hydrolysis system. Therefore, 2InaQ-N-CSN46A had higher hydrolysis efficiency and stability than InaQ-N-CSN46A. GPC revealed that under the optimum enzymatic hydrolysis temperature, the final products were mainly chitobiose and chitotriose. Chitopentaose accumulated (77.62%) when the hydrolysis temperature reached 60 ℃. FTIR and NMR analysis demonstrated that the structures of the two hydrolysis products were consistent with those of COSs.Conclusions In this study, chitosanase was expressed on the surfaces of E. coli by increasing induction temperature, and chitosan was hydrolysed directly without enzyme purification steps. This study provided a novel strategy for industrial COSs production.


2008 ◽  
Vol 74 (11) ◽  
pp. 3342-3348 ◽  
Author(s):  
Masayoshi Tanaka ◽  
Yuko Nakata ◽  
Tetsushi Mori ◽  
Yoshiko Okamura ◽  
Hitoshi Miyasaka ◽  
...  

ABSTRACT Bacterial cell surface display is a widely used technology for bioadsorption and for the development of a variety of screening systems. Magnetotactic bacteria are unique species of bacteria due to the presence of magnetic nanoparticles within them. These intracellular, nanosized (50 to 100 nm) magnetic nanoparticles enable the cells to migrate and be manipulated by magnetic force. In this work, using this unique characteristic and based on whole-genomic and comprehensive proteomic analyses of these bacteria, a cell surface display system has been developed by expressing hexahistidine residues within the outer coiled loop of the membrane-specific protein (Msp1) of the “Magnetospirillum magneticum” (proposed name) AMB-1 bacterium. The optimal display site of the hexahistidine residues was successfully identified via secondary structure prediction, immunofluorescence microscopy, and heavy metal binding assay. The established AMB-1 transformant showed high immunofluorescence response, high Cd2+ binding, and high recovery efficiency in comparison to those of the negative control when manipulated by magnetic force.


2009 ◽  
Vol 76 (3) ◽  
pp. 971-973 ◽  
Author(s):  
Jong Hwan Baek ◽  
Mee-Jung Han ◽  
Seung Hwan Lee ◽  
Sang Yup Lee

ABSTRACT A cell surface display system was developed using Escherichia coli OmpC as an anchoring motif. The fused Pseudomonas fluorescens SIK W1 lipase was successfully displayed on the surface of E. coli cells, and the lipase activity could be enhanced by the coexpression of the gadBC genes identified by transcriptome analysis.


Yeast ◽  
2002 ◽  
Vol 19 (13) ◽  
pp. 1153-1163 ◽  
Author(s):  
So-Young Kim ◽  
Jung-Hoon Sohn ◽  
Yu-Ryang Pyun ◽  
Eui-Sung Choi

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