Expression,Purification,Characteristics and Homology Modeling of the HMGS from Streptococcus pneumoniae

Objective To understand the molecular basis for a potential reaction mechanism and develop novel antibiotics with homology modeling for 3-hydroxy-3-methylglutaryl coenzyme A （HMG-CoA） synthase （HMGS）. Methods The genetic engineering technology and the composer module of SYBYL7.0 program were used, while the HMGS three-dimensional structure was analyzed by homology modeling. Results The mvaS gene was cloned from Streptococcus pneumoniae and overexpressed in Escherichia coli from a pET28 vector. The expressed enzyme （about 46 kDa） was purified by affinity chromatography with a specific activity of 3.24 μmol/min/mg. Optimal conditions were pH 9.75 and 10 mmol/L MgCl2 at 37 ℃ The Vmax and Km were 4.69 μmol/min/mg and 213 μmol/L respectively. The 3D model of S.pneumoniae HMGS was established based on structure template of HMGS of Enterococcus faecalis. Conelusion The structure of HMGS will facilitate the structure-based design of alternative drugs to cholesterol-lowering therapies or to novel antibiotics to the Gram-positive cocci, whereas the recombinant HMGS will prove useful for drug development against a different enzyme in the mevalonate pathway.

ANALYSES OF ERRORS IN MEDIUM-TERM NUMERICAL FORECAST PRODUCTS FOR THE SUBTROPICAL HIGH 1998

By statistic and dynamic analyses, we have come to the following conclusions: (1) The ECMWF medium-term numerical forecast can forecast medium-term activity of subtropical high, and the accuracy rate of forecast cannot have large improvement by translational corrections. (2) The important cause for the ECMWF medium-term numerical forecast to have errors in 1998 is that the astronomical tide is not included in the model. (3) Two indexes are found from which it can be judged that ECMWF medium-term numerical forecast will have errors if the astronomical tide is ignored in the model : ① When the 54.7?line under the moon of the nodical month astronomical singularities coincides with the trough-line of the subtropical jet flow from 50癊 to 150癊 on the 500 hPa level at 2000 L.T. of the same day, and is approximately vertical (α>60? with the isotherm, then the day 0 2 days after the appearance of the nodical month astronomical singularities is defined as the initial day. Then in three successive days after the initial day, ECMWF medium-term numerical forecast of the northern latitude of the 588 line at 120 癊 will have continuous errors as large as two latitudes (7/9). Otherwise, it won’t have continuous errors (13/13). ② Otherwise, if the 54.7 ?line is in the range of a low pressure between two high pressures, then there is a dispersive error on the day of the nodical month astronomical singularities (5/7). There is not any error (6/6) otherwise.