Production,Purification and Characterization of the Hen Egg-White Lysozyme Inhibitor from Enterobacter cloacae M-1002

Three hen egg-white lysozyme inhibitor producing strains, Enterobacter cloacae M-1002, E. sakazakii M-1204, and Erwinia rhapontici H-55, were isolated from the soils of Taiwan. E. cloacae M-1002 appeared to be a promising inhibitor producing strain. One inhibitor was isolated from the culture broth of this strain. Maximum lysozyme inhibitory activity was obtained when the bacterium was grown aerobically in a medium consisting of 0.75% glucose, 0.25% beef extract, 1.0% polypeptone, and 0.25% sodium L-glutamate (pH 70) at 37 °C after 36–48 hrs. A hen egg-white lysozyme inhibitor was isolated from the culture broth of this strain. The inhibitor was purified from the culture supernatant of E. cloacae M-1002 by ammonium sulfate fractionation, DEAE-Sepharose CL-6B column chromatography and Fractogel TSK HW-55 (S) gel chromatography. Molecular weight of the purified lysozyme inhibitor was estimated to be 18, 000–20, 000 by SDS-PAGE and HPLC, and was composed of 71% amino acid and 23% total sugar. Serine, glycine, and alanine in a 3:2:1 molar ratio were the major amino acids, calculated to be 32.8, 20.3, and 11.4% (mol%), respectively. Glucose and mannose were the major sugar components of the inhibitor. The inhibitor was stable at pH 5 to 8 and was stable under 50 °C. Only hen egg-white lysozyme was inhibited by the purified inhibitor but not the other tested enzymes such as lysozyme of celery, turnip; lytic enzyme of Pseudomonas aeruginosa M-1001; chitinase/lysozyme of P. aeruginosa K-187; or cellulase and xylanase of Streptomyces actuosus A-151 and Aspergillus sp. G-393. The inhibition of lysozyme to the bacterial cell lytic activity by the purified inhibitor was 100%.     

Holographic interferometry as electrochemical emission spectroscopy of a carbon steel in 5–20 ppm TROS C-70 inhibited seawater

In the present investigation, holographic interferometry was utilized for the first time to determine the rate change of the number of the fringe evolutions during the corrosion test of a carbon steel in blank seawater and seawater with different concentrations of a corrosion inhibitor. In other words, the anodic dissolution behaviors (corrosion) of the carbon steel were determined simultaneously by holographic interferometry, as an electromagnetic method, and by the electrochemical impedance (EI) spectroscopy, as an electronic method. So, the abrupt rate change of the number of the fringe evolutions during corrosion tests, EI spectroscopy, of the carbon steel is called electrochemical emission spectroscopy. The corrosion process of the steel samples was carried out in blank seawater and seawater with different concentrations, 5–20 ppm, of TROS C-70 corrosion inhibitor using the EI spectroscopy method, at room temperature. The electrochemical-emission spectra of the carbon steel in different solutions represent a detailed picture of the rate change of the anodic dissolution of the steel throughout the corrosion processes. Furthermore, the optical interferometry data of the carbon steel were compared to the data, which was obtained from the EI. spectroscopy. Consequently, holographic interferometry is found very useful for monitoring the anodic dissolution behaviors of metals, in which the number of the fringe evolutions of the steel samples can be determined in situ.     

Glycopolymeric inhibitors of β‐glucuronidase. III. Configurational effects of hydroxy groups in pendant glyco‐units in polymers upon inhibition of β‐glucuronidase

Two kinds of new glycopolymers, (P(VB‐1‐GlcaH‐co‐AAm), 9 ) and (P(VB‐1‐Glco‐co‐AAm), 10 ), were synthesized through the radical copolymerization of styrene derivatives bearing pendant D ‐glucaric and D ‐gluconic moieties, N‐(p‐vinylbenzyl)‐1‐D ‐glucaramide (VB‐1‐GlcaH, 7 ), and N‐(p‐vinylbenzyl)‐D ‐gluconamide (VB‐1‐Glco, 8 ), with acrylamide (AAm). Glycopolymer 9 bearing the pendant glucaric moiety at the first position inhibited the hydrolysis of a model compound for xenobiotics‐β‐glucuronide conjugates, p‐nitrophenyl β‐D ‐glucuronide, uncompetitively, in contrast to the competitive inhibition in the presence of the corresponding isomeric glycopolymer bearing the pendant D ‐glucaric unit at the sixth position (P(VB‐6‐GlcaH‐co‐AAm), 3 ) reported in our previous article. On the other hand, another copolymer 10 bearing the gluconic moiety was found not to inhibit the hydrolysis as well as the corresponding copolymer bearing pendant gulonic unit (P(VB‐6‐Glco‐co‐AAm), 4 ). These results indicate that the hydrolysis is influenced not only by existence of pendant carboxyl units but also by the direction on the linkage of the glyco‐units to the polymer frame. Therefore the configurational position of hydroxy groups in pendant glyco‐units in macromolecular inhibitors may be essential for the interaction with β‐glucuronidase. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 4895–4903, 2006     

铜表面腐蚀的激光扫描微区光电压图象的研究

铜表面腐蚀的激光扫描微区光电压图象的研究杨迈之，张雯，蔡生民，任聚杰（北京大学化学系，北京，１０００７１）（河北医学院化学系，石家庄）潘传智，杨勇（浙江工业大学化工系，杭州）（厦门大学化学系，厦门）关键词铜，缓蚀剂，微区光电压图铜的腐蚀与防腐早已引起...     

Studies toward the synthesis of salinosporamide A, a potent proteasome inhibitor

An α-methylenepyrrolidinone bearing all the functionalities and relative configurations of an advanced intermediate in the synthesis of salinosporamide A and analogues has been synthesized from methyl pyroglutamate through regio- and stereoselective N-methylnitrone cycloaddition.     

Development of a capillary electrophoretic method for the separation of diastereoisomers of a new human immunodeficiency virus protease inhibitor

A capillary electrophoretic (CE) method was developed for the separation of diastereoisomers of a new human immunodeficiency virus (HIV) protease inhibitor TMC114. In total 16 isomers of this drug have been synthesized (eight pairs of enantiomers). We succeeded in the separation of the eight diastereoisomers, but no enantiomers could be separated. Because of the high similarity and water-insolubility of these isomers, the separation is a real challenge. Different CE modes were tried out: capillary zone electrophoresis (CZE), nonaqueous capillary electrophoresis (NACE), micellar electrokinetic capillary chromatography (MEKC), and microemulsion electrokinetic capillary chromatography (MEEKC). Only MEEKC offered resolution of these compounds.     

Gold nanoparticle-enhanced surface plasmon resonance measurement with a highly sensitive quantification for human tissue inhibitor of metalloproteinases-2

Two different methods for the quantification of human tissue inhibitor of metalloproteinases-2 (TIMP-2) were developed using surface plasmon resonance (SPR) and gold nanoparticles for signal enhancement. The first method, a competitive assay, used TIMP-2 immobilized to the sensor surface and the inactive form of matrix metalloproteinase-2 (proMMP-2) (EC 3.4.24.24) adsorbed to gold nanoparticles. The sensor signals resulting from the interaction of MMP-2-gold nanoparticles with immobilized TIMP-2 were inversely proportional to the amounts of TIMP-2 of the sample. The measuring range for TIMP-2 was about 15–180 pM. The second method, a one-step sandwich assay, used proMMP-2 immobilized to the sensor surface and an anti-TIMP-2 monoclonal antibody coupled to gold nanoparticles. The lower detection limit of this assay format was 0.5 pM of TIMP-2. The binding signals were highly reproducible up to 100 pM of the inhibitor. The improvements obtained in TIMP-2 quantification over already existing tests could contribute to a better understanding and diagnosis of diseases like cancer.     

New bioactive triterpenoid from Oldenlandia cantonensis How

A new pentacyclic triterpenoid, urs-12-en-29α-oic acid-3β-ol (1), was obtained from the ethanol extract of Chinese herb Oldenlandia cantonensis How. The structure was elucidated by spectroscopy methods, including nuclear magnetic resonance (NMR) (1D and 2D), infrared spectroscopy (IR), and mass spectrometry (MS). 1 exhibited significant inhibitory activity against the human DNA topoisomerase I (hTopo I), the cancer cell lines BEL-7402 and MCG-803, with the IC₅₀ values 12.0, 6.5, and 8.0 μg/mL, respectively. The volatile oil, the fraction of petroleum ether: EtOAc = 20:1 (V/V) on Si gel chromatography, was also quantitatively analyzed by gas chromatography mass spectrometry (GC-MS). As a result, 60 compounds were identified. Among them, the long chain aliphatics, terpenes and steroids, as the representative structure type, were found with percentages of 36.16%, 6.42% and 9.28%, respectively. Translated from Chinese Journal of Applied Chemistry, 2006, 23(8): 871–874 [译自: 应用化学]     

Synthesis and kinetic evaluation of 4-deoxy-4-phosphonomethyl-d-erythronate, the first hydrolytically stable and potent competitive inhibitor of ribose-5-phosphate isomerase

4-deoxy-4-Phosphonomethyl-d-erythronate, an isosteric and hydrolytically stable analogue of the known ribose-5-phosphate isomerase inhibitor 4-deoxy-4-phospho-d-erythronate, was obtained by a 14-step synthesis from d-arabinose through an highly improved synthesis of the precursor 5-deoxy-5-phosphonomethyl-d-arabinose. The title compound appears as the first stable and potent competitive inhibitor of the enzyme catalyzed isomerization of ribose-5-phosphate to d-ribulose-5-phosphate (K_i=74 μM, K_m/K_i=100), exhibiting only a 3-fold weaker inhibitory activity than its phosphate analogue.     

Synthesis of PI-88 analogue using novel O-glycosidation of exo-methylenesugars

Acid-promoted α-stereoselective O-glycosidation of 1-exo-methylenesugars was successfully applied to the synthesis of a PI-88 analogue. By using methanesulfonic acid as a promoter, 1′-C-methyl-α-disaccharides with p-methoxybenzyl protection were obtained in high yield. The sequence of selective deprotection and glycosidation provided 1-C-methyl-pentasaccharide efficiently.