Send to

Choose Destination
Wei Sheng Wu Xue Bao. 2009 Feb;49(2):191-7.

Glycine-aspartic_ acid-serine-leucine esterase Xcc_ est from Xanthomonas campestris pv. campestris 8004 and its esterase domain: gene expression in Escherichia coli, refolding and characterization.

Author information

State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.



To characterize the GDSL (glycine, aspartic acid, serine and leucine motif in protein sequence) esterase Xcc_ est from Xanthomonas campestris pv. campestris (Xcc) 8004.


Xcc_ est gene and different domains of Xcc_ est gene were PCR amplified and expressed in Escherichia coli, the HIS-Tagged fusion proteins were purified by Ni-NTA chromatography.


The optimum pH and temperature of partly purified Xcc_ est were 8.0 and 52 degrees C when pNPB (4-nitrophenylbutyrate) was used as substrate. The Km and Vmax value of Xcc_ est and the passenger domain (Xcc_ estN1-334) for pNPB were 47.6 +/- 4.6 mol/L, 67.6 +/- 7.8 U/mg and 469.4 +/- 9.8 mol/L, 2.5 +/- 0.9 U/mg respectively. Inclusion bodies of mature domain Xcc_ est (Xcc_ estN26-606) could be refolded but inclusion bodies of the passenger domain (Xcc_ estN26-334) could not be refolded. Refolded mature domain had broad substrate spectrum and showed higher stability than Xcc_ est when stored at 25 degrees C.


Refolded Xcc_ estN26-606 can be a candidate for biotransformation application.

[Indexed for MEDLINE]

Supplemental Content

Loading ...
Support Center