The entire gene of carboxyltransferase（CT） domain of acetyl-CoA carboxylase（ACCase） from Chinese Spring wheat（CSW） plastid was cloned firstly, and the 2.3 kb gene was inserted into PET28a^＋ vector and expressed in E. coil in a soluble state. The （His）6 fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass（Mr） was 88000. The results of the sequence analysis indicate that the cloned gene（GeneBank accession No. EU124675） was a supplement and revision of the reported ACCase CT partial cDNA from Chinese Spring wheat plastid. The recombinant protein will be significant for us to investigate the recognizing mechanism between ACCase and herbicides, and further to screen new herbicides.
WANG Rui-jianYANG Xue-yingZHENG Liang-yuYANG YeGAO GuiCAO Shu-gui
To identify the desired hypertherrnophilic variants within a mutant esterase library for the resolution of （R, S）-2- octanol acetate, a simple, reliable, and versatile method was developed in this study. We built a screening strategy including two steps, first we selected agar plate with substrate to screen the enzymatic activity; secondly we used a pH indicator to screen the enantioselectivity. This method could rapidly detect favorable mutants with high activity and enantioselectivity. A total of 96. 2% of tedious screening work can be precluded using this screening strategy. It is an effective screening for alkyl ester and can be applied to relative screening researches. The four improved mutants were screened from the mutant esterase library. Their enantioselectivities, activities, and structures were investigated at different temperatures.
ZHANG Gui-rong GAO Ren-jun ZHANG Ai-jun RAO Lang CAO Shu-gui
Introduction Nerve growth factor （NGF） was first discovered and purified by Rita Levi-Montalcini and Stanley Cohen in the 1950s. It represents the first cellular growth factor ever discovered and involved in the gl-owth ,survival, and differentiation of specific nerve cell populations. Although animal tests and trials indicate that rhNGF could be ment for diabetic and HIV-related phase- Ⅱ clinical an effective treatneuropathies , a large-scale phase-Ⅲ clinical trial has failed to give similar result. NGF isolated from the mouse submaxillary gland has been widely used clinically in China for the treatment of peripheral neuropathy,
The changes in the activity and the conformation of the hyperthermophilic esterase derived from aerobic thermophilic Aeropyrum pernix K1 （ APE1547 ） were studied during denaturation by guanidine hydrochlofide （ GdnHC1 ） and urea. The denaturation course of APE1547 was followed by the steady-state and time resolved fluorescence methods. An increase in the denaturant concentration in the denatured system can significantly enhance the inactivation and unfolding of APE1547. The enzyme can be completely inactivated with a urea concentration of 2.7 mol/L or a GdnHCl concentration of 7.5 mol/L. The fluorescence emission maximum of the enzyme protein red shifts in magnitude to a maximum value（355 nm） when the concentration of GdnHCl is 5.1 mol/L. The experimental results indicate that APE1547 has a high resistance to urea. Unfolding of APE1547 in GdnHCI（4. 2-6.0 mol/L） was shown to be an irreversible process. The present results indicate that the ion pairs in this protein may be a key factor for the stability of this esterase.
