Biomedical subjects

Jian Wei Liu

Publications and source records attributed to Jian Wei Liu.

3 recordsLinked to original sources

PubMed2006

The purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes.

The metallo-glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) has been cloned, expressed in Escherichia coli and purified. Initial screening of crystallization conditions for this enzyme resulted in the identification of needles from one condition in a sodium malonate grid screen. Removal of the metals from the enzyme and subsequent optimization of these conditions led to crystals that diffracted to 2.9 angstroms and belonged to space group P2(1)3, with unit-cell parameter a = 164.1 angstroms. Self-rotation function analysis and V(M) calculations indicated that the asymmetric unit contains two copies of the monomeric enzyme, corresponding to a solvent content of 79%. It is intended to determine the structure of this protein utilizing SAD phasing from transition metals or molecular replacement.

Bacterial Proteins↗

PubMed2006

Fluoride levels in various black tea commodities: measurement and safety evaluation.

In order to determine the fluoride content of various products of black tea and to make safety evaluations, 37 commodities in the different manufactured forms of tea were assessed by means of ion-selective electrode and spectrophotometric methods. The results showed wide differences in fluoride levels. The fluoride content was found 0.95-1.41 mg/L in black tea sticks, 0.70-2.44 mg/L in black tea granules and 1.15-6.01 mg/L in black tea bags. Of the products tested, the fluoride content was greatest in black tea bags, presumably because black tea bags are made of low cost, and older tea leaves. According to WHO's recommendation, fluoride exposure threshold for children is 2mg of daily intake and for adult, 4 mg. Assuming that children's daily intake of black tea was 800 ml and adult's 1500 ml, 56% of black tea bags should be considered unsafe for children and 44% should be considered unsafe for adults. At average rates of consumption, 24% of the black tea bags could result in an increasing risk of osteofluorosis and fractures (WHO). In the areas with drinking water fluoridation or other fluoride supplements, excessive fluoride exposure can become even more severe with even lower intake tea made from black tea bags. The potential hazards of fluoride arising from consumption of black tea has not aroused due public attention. Therefore, it is necessary to establish reliable fluoride surveillance in food and beverage commodities and conduct additional epidemiological research on fluoride hazards. In addition, it is urgent that governmental and international agencies adopt safe standards of fluoride content in tea commodities.

Electrochemistry↗

PubMed2005

Following directed evolution with crystallography: structural changes observed in changing the substrate specificity of dienelactone hydrolase.

The enzyme dienelactone hydrolase (DLH) has undergone directed evolution to produce a series of mutant proteins that have enhanced activity towards the non-physiological substrates alpha-naphthyl acetate and p-nitrophenyl acetate. In terms of steady-state kinetics, the mutations caused a drop in the K(m) for the hydrolysis reaction with these two substrates. For the best mutant, there was a 5.6-fold increase in k(cat)/K(m) for the hydrolysis of alpha-naphthyl acetate and a 3.6-fold increase was observed for p-nitrophenyl acetate. For alpha-naphthyl acetate the pre-steady-state kinetics revealed that the rate constant for the formation of the covalent intermediate had increased. The mutations responsible for the rate enhancements map to the active site. The structures of the starting and mutated proteins revealed small changes in the protein owing to the mutations, while the structures of the same proteins with an inhibitor co-crystallized in the active site indicated that the mutations caused significant changes in the way the mutated proteins recognized the substrates. Within the active site of the mutant proteins, the inhibitor was rotated by about 180 degrees with respect to the orientation found in the starting enzyme. This rotation of the inhibitor caused the displacement of a large section of a loop on one side of the active site. Residues that could stabilize the transition state for the reaction were identified.

Binding Sites↗