Evaluation of enantioselectivity in lipase-catalyzed acylation of hydroxyalkylphosphine oxides

The lipase-catalyzed optical resolution of 2-, 3-, and 5-hydroxyalkyl phosphorus compounds 1 provided the corresponding optically pure diastereomers in good yields. (S_P, R)- and (R_P, S)-1 were acylated faster than (S_P, S)- and (R_P, R)-1. The stereoselectivity at the phosphorus atom changed with the flexibility of the active sites in the lipases. The stereoselectivity at the phosphorus atom was higher in the reaction of 1a than in the reaction of 1b,c. The reaction rate of -hydroxyalkylphosphine oxide 1c was faster than that of 1a, although less enantioselectivity was observed at the phosphorus atom.     

One step purification of Escherichia coli beta-glucuronidase

beta-glucuronidase was purified by affinity chromatography on thiophenyl-glucuronide coupled to Sepharose. The enzyme was more than 95% pure. This enzyme is a tetramer composed of identical 74 kDa monomers. The amino-terminal sequence determined was: NH2-Met-Leu-Arg-Pro-Val.     

水稻土中铁氧化物对产甲烷古菌群落结构的影响

产甲烷菌广泛分布在淹水水稻土等各种厌氧环境中,在全球气候变化、碳循环和能源等领域都发挥着重要的作用。研究发现,厌氧条件下,水稻土中铁氧化物的生物还原会抑制产甲烷菌的甲烷合成作用。然而,目前关于铁氧化物对产甲烷菌群落结构的影响报道较少。通过泥浆厌氧培养实验,向采集的水稻土中添加甲酸盐作为甲烷合成的底物(Control,CK处理),并设置添加水铁矿作为体系中唯一电子受体的处理组(Ferrihydrite,Fh处理)。培养结束后,与CK相比,添加水铁矿显著降低了古菌在总微生物群落中的占比,但对古菌群落的物种多样性和均一度没有显著影响;且两组处理中优势种均为操作分类单元(Operational taxonomic unit,OTU)2056和OTU 911(76%—80%)。这说明碳源相同时,产甲烷菌的群落结构不受铁氧化物的影响。本研究为探索土壤中微生物铁还原与碳循环耦合的分子机制奠定基础。     

Biocatalysis of nitro substituted styrene oxides by non-conventional yeasts

Yeast strains (410) from more than 45 different genera were screened for the enantioselective hydrolysis of nitro substituted styrene oxides. These strains included 262 yeasts with known epoxides hydrolase activity for various other epoxides. Epoxide hydrolase activity for p-nitrostyrene oxide (pNSO) (177 strains) and m-nitrostyrene oxide (mNSO) (148 strains) was widespread in the yeasts, while activity for o-nitrostyrene oxide (oNSO) was less ubiquitous (22 strains). The strains that displayed enantioselectivity in the hydrolysis of one or more of the nitro substituted styrene oxides (35 strains) were also screened against styrene oxide (SO). Rhodosporidium toruloides UOFS Y-0471 displayed the highest enantioselectivity for pNSO (ee 55%, yield 35%) while Rhodotorula glutinis UOFS Y-0653 displayed the highest enantioselectivity for mNSO (ee >98%, yield 29%), oNSO (ee 39%, yield 19%) and SO (ee >98%, yield 19%). (R)-Styrene oxide was preferentially hydrolysed to the corresponding (R)-diol with retention of configuration at the stereogenic centre. In the case of the nitro substituted styrene oxides the absolute configurations of the remaining epoxides and the formed diols were not established.     

Inhibitory effects of nitrogen oxides on a mixed methanogenic culture

The effect of nitrate, nitrite, nitric oxide (NO), and nitrous oxide on a mixed, mesophilic (35 degrees C) methanogenic culture was investigated. Short-term inhibition assays were conducted at a concentration range of 10-350 mg N/L nitrate, 17-500 mg N/L nitrite, 0.02-0.8 mg N/L aqueous NO, and 19-191 mg N/L aqueous nitrous oxide. Simultaneous methane production and N-oxide reduction was observed in 10 and 30 mg N/L nitrate and 0.02 mg N/L aqueous NO-amended cultures. However, addition of N-oxide resulted in immediate cessation of methanogenesis in all other cultures. Methanogenesis completely recovered subsequent to the complete reduction of N-oxides to nitrogen gas in all N-oxide-amended cultures, with the exception of the 500 mg N/L nitrite- and 0.8 mg N/L aqueous NO-amended cultures. Partial recovery of methanogenesis was observed in the 500 mg N/L nitrite-amended culture in contrast to complete inhibition of methanogenesis in the 0.8 mg N/L aqueous NO-amended culture. Accumulation of volatile fatty acids was observed in both cultures at the end of the incubation period. Among all N-oxides, NO exerted the most and nitrate exerted the least inhibitory effect on the fermentative/methanogenic consortia. The effect of multiple additions of nitrate (300 mg N/L) on the same methanogenic culture was also investigated. Long-term exposure of the methanogenic culture to nitrate resulted in an increase of N-oxide reduction rates and decrease of methane production rates, which was attributed to changes in the microbial community structure due to nitrate addition.     

One step purification of chymosin by mixed mode chromatography

Mixed mode Sepharose and Perloza bead cellulose matrices were prepared using various chemistries. These matrices contained hydrophobic (aliphatic and/or aromatic) and ionic (carboxylate or alkylamine) groups. Hydrophobic amine ligands were attached to epichlorohydrin activated Sepharose (mixed mode amine matrices). Hexylamine, aminophenylpropanediol and phenylethylamine were the preferred ligands, on the basis of cost and performance. Other mixed mode matrices were produced by incomplete attachment (0-80%) of the same amine ligands to carboxylate matrices. The best results were obtained using unmodified or partially ligand-modified aminocaproic acid Sepharose and Perloza. High ligand densities were used, resulting in high capacity. Furthermore, chymosin was adsorbed at high and low ionic strengths, which reduced sample preparation requirements. Chymosin, essentially homogeneous by electrophoresis, was recovered by a small pH change. The methods described were simple, efficient, inexpensive and provided very good resolution of chymosin from a crude recombinant source. The carboxylate matrices had the best combination of capacity and regeneration properties. The performance of Sepharose and Perloza carboxylate matrices was similar, but higher capacities were found for the latter. Because it is cheaper and can be used at higher flow rates, Perloza should be better suited to large scale application. High capacity chymosin adsorption was found with carboxymethyl ion exchange matrices, but low ionic strength was essential for adsorption and the purity was inferior to that of the mixed mode matrices. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 45-55, 1997.     

Gram-scale chromatographic purification of beta-sitosterol. Synthesis and characterization of beta-sitosterol oxides

An effective purification method for beta-sitosterol was developed starting from a commercial source of a phytosterol mixture using preparative adsorption column chromatography. beta-Sitosterol (> or = 95% purity) was obtained on a gram-scale. Thus, the synthesis of six beta-sitosterol oxides, including 7alpha-hydroxy, 7beta-hydroxy, 5,6alpha-epoxy, 5,6beta-epoxy, 7-keto, and 5alpha,6beta-dihydroxysitosterol, were successfully carried out. The spectral characteristics of all the synthetic intermediates and target compounds (approximately 95% purity) were well-documented.     

Relationship between Pb/Cd adsorption and metal oxides on surface coatings at different depths in Lake Jingyuetan

Adsorption of heavy metals to metal oxides on surface coatings of sediments in aquatic environments is one of the most important factors governing their toxicity. Many previous studies were carried out to examine relationships between characteristics of surface coatings and heavy metal adsorption. However, a number of uncertainties existed in the related mechanisms. Especially, the effect of depth, at which surface coatings are developed, was not seriously considered. In this study, surface coatings were collected on glass slides at different depths in the Jingyuetan Lake, which is located in the northeast of China, and the related chemical characteristics (Fe and Mn oxides in the surface coatings) were analyzed. Pb and Cd adsorption onto the surface coatings was measured under controlled laboratory conditions. Nonlinear regression analyses and Langmuir adsorption isotherms were used to estimate contributions of Fe and Mn oxides developed at different depths. The results indicated that a strong linear relationship existed between the depth of water and the contents of iron/manganese oxides. The depth of water can also influence the Langmuir parameters (_max) of Pb and Cd adsorbed onto the surface coatings, by reducing values of _max from water surface to the bottom and reaching the lowest level when approaching the sediments. For surface coatings at different depths in the lake, analyses of correlation between the _max and the coating constituents indicated that adsorption of Pb and Cd by Mn and Fe oxides are statistically significant. Based on results of this study, the role of water depth in governing processes of Pb and Cd adsorption to metal oxides in homogeneous surface coatings was verified.     

Solubility and dissolution of iron oxides

In most soils, FeIII oxides (group name) are the common source of Fe for plant nutrition. Since this Fe has to be supplied via solution, the solubility and the dissolution rate of the Fe oxides are essential for the Fe supply. Hydrolysis constants and solubility products (K_sp) describing the effect of pH on FeIII ion concentration in solution are available for the well-known Fe oxides occurring in soils such as goethite, hematite, ferrihydrite. K_sp values are usually extremely low ((Fe³⁺)·(OH)³=10^–37–10^–44). However, for each mineral type, K_sp may increase by several orders of magnitude with decreasing crystal size and it decreases with increasing Al substitution assuming ideal solid solution between the pure end-members. Based on such calculations a poorly crystalline goethite with a crystal size of 5 nm may well reach the solubility of ferrihydrite. The variations in K_sp are of relevance for soils because crystal size and Al substitution of soil Fe oxides vary considerably and can now be determined relatively easily.The concentration of Fe²⁺ in soil solutions is often much higher than that of Fe(III) ions. Therefore, redox potential strongly influences the activity of FeII. At a given pH and E_h, the activity of FeII is higher the higher K_sp of the FeIII oxide and thus also varies with the type of Fe oxide present.Besides the solubility, it is the dissolution rate which governs the supply of soluble Fe to the plant roots. Dissolution of Fe oxides takes place either by protonation, complexation or, most important, by reduction. Numerous dissolution rate studies with various FeIII oxides were conducted in strong mineral acids (protonation) and they have shown that besides the Fe oxide species, crystal size and/or crystal order and substitution are important determinative factors. For example, in soils, small amounts of a more highly soluble meta- or instable Fe oxide such as ferrihydrite with a large specific surface (several hundred m²g^–1) may be essential for the Fe supply to the plant root. Its higher dissolution rate can also be used to quantify its amount in soils. Ferrihydrite can be an important component in soils with high amounts of organic matter and/or active redox dynamics, whereas highly aerated and strongly weathered soils are usually very low in ferrihydrite. On the other hand, dissolution rates of goethites decrease as their Al substitution increases.Much less information exists on the rate of reductive and chelative dissolution of Fe oxides which generally simulate soil conditions better than dissolution by protonation. Here again, type of oxide, crystal size and substitution are important factors. Organic anions such as oxalate, which are adsorbed at the surface, may weaken the Fe³⁺-O bonds and thereby increase reductive dissolution. As often observed in weathering, the dissolution features of the crystals appear to follow zones of weakness in the crystal.     

外源氧化铁对水稻土中有机酸含量的影响

在水稻土泥浆中添加Fe(OH) 3 可显著降低乙酸浓度 .在新鲜水稻土样品中 ,由于添加Fe(OH) 3导致对乙酸的竞争消耗 ,在培养 5d后 ,乙酸浓度降至 10～ 2 0 μmol·L-1的稳态浓度 ,而此刻对照中的乙酸浓度仍在 12 0 0 μmol·L-1以上 .在乙酸产生量较低的土壤中 ,添加Fe(OH) 3 可完全消耗体系中的乙酸 ,并导致产CH4过程的完全被抑制 .添加纤铁矿同样可使乙酸浓度显著减少 ,但作用效果不如无定形氧化铁 .添加赤铁矿可造成培养初期 (10d以内 )乙酸的大量积累 ,但并不引起产CH4量的增大 .添加Fe(OH) 3 、纤铁矿及铝取代针铁矿 ,能引起厌氧培养的水稻土中丙酸浓度的降低 ,其抑制效率为Fe(OH) 3 >纤铁矿 >铝取代针铁矿 .新鲜土样和经过 11周厌氧处理后的土样中 ,有机酸种类和含量有较大差别 .     

Synthesis and characterization of new imidophosphanes and phosphine oxides containing 3,3,4,4-tetramethylsuccinimidyl group

A series of new imidophosphanes and phosphine oxides containing 3,3,4,4-tetramethylsuccinimidyl group were synthesized and characterized by ¹H, ¹³C{¹H} and ³¹P NMR spectroscopy, IR and MS. Ph_mPCl_n (m = 3 − n, n = 3, 2, 1) reacted with 3,3,4,4-tetramethylsuccinimide (TH) and potassium 3,3,4,4-tetramethylsuccinimidate 1 to give corresponding Ph_mPT_n. Molecular structures of products were established by single-crystal X-ray diffraction experiments. Attempts to prepare new imidophosphoranes by reactions of 1 with Ph_mPCl_n (m = 5 − n, n = 4, 3, 2) resulted in phosphine oxides. In these reactions the phosphoryl group was formed and we characterized a by-product of this type of reaction.     

One step flow-through adsorptive purification of tubulin from tissue homogenate

Tubulin, a potential target for anti-cancer drugs, has been purified in one step and obtained as flow-through fraction directly from an extract of a mammalian brain tissue by adsorption chromatography on H-CELBEADS, an indigenously developed rigid, superporous cross-linked cellulose based weakly hydrophobic adsorbent. The fibrous polymerized tubulin mass passed through the H-CELBEADS bed while the associated proteins were separated by adsorption. The final tubulin preparation was obtained free from other proteins as seen on SDS-PAGE. Purified tubulin was obtained in a yield of about 29 mg/100 g brain, and its bioactivity, evaluated through its ability to bind colchicine, was found to be preserved.     

One step towards restoration of self-tolerance in human autoimmune diseases

In developed countries the incidence of autoimmune insulin-dependent or type 1 diabetes as the one of all autoimmune diseases has steadily increased over the last decades. Conventional therapy of type 1 diabetes is essentially palliative namely, chronic delivery of exogenous insulin that is associated with major constraints (multiple daily parenteral administration, serious risks linked to hypoglycemic episodes) and incomplete effectiveness in preventing severe degenerative complications. This explains the growing attention on modern therapeutic strategies using biological agents such as CD3 monoclonal antibodies that allow 'reprogramming' the immune system to restore self-tolerance to pancreatic beta cell antigens. This strategy which proved successful in the experimental setting has recently been translated to the clinic with very encouraging results. CD3 antibodies may represent a new category of drugs affording a real cure for autoimmunity namely, inhibiting the pathogenic immune response while preserving the host reactivity to unrelated antigens.     

Preparation and characterization of chitosans carrying aldehyde functions generated by nitrogen oxides

In this work, aldehyde-functionalized chitosan is produced by the reaction of chitosan with nitrogen oxides generated in situ from a HNO₃/H₃PO₄-NaNO₂ mixture. This method is more advantageous than the existing approaches, since the depolymerization is slower and the purification process is straightforward. The appearance of characteristic peaks in the Fourier transform infrared and carbon-13 nuclear magnetic resonance spectra (1733 cm⁻¹ and 183.4 ppm, respectively) of the product confirms the presence of the aldehyde functionality in the modified chitosans. The ¹H NMR spectra also revealed the presence of aldehyde groups. Furthermore, the gradual disappearance of the peaks due to aldehyde protons and a concomitant appearance of a new resonance at ∼8.05 ppm with increasing pH indicate the formation of Schiff's base between the aldehyde and the free amine groups. The aldehyde-functionalized chitosan prepared with 6 h of reaction time (chitosan-6h) forms a gel in situ without any added external crosslinker and it may potentially be useful as a vehicle for drug delivery.     

Nanostructured copper oxides and phosphates from a new solid-state route

Nanostructured copper containing materials of CuO, Cu₃(PO₄)₃ and Cu₂P₂O₇ have been prepared by solid-state pyrolysis of molecular CuCl₂·NC₅H₄OH (I), CuCl₂·CNCH₂C₆H₄OH (II), oligomeric [Cu(PPh₃)Cl]₄ (III), N₃P₃[OC₆H₄CH₂CN·CuCl]₆[PF₆] (IV), N₃P₃[OC₆H₅]₅[OC₅H₄N·Cu][PF₆] (V), polymeric chitosan·(CuCl₂)_n (VI) and polystyrene-co-4-vinylpyridine PS-b-4-PVP·(CuCl₂) (VII) precursors. The products strongly depend on the precursor used. The pyrolytic products from phosphorus-containing precursors (III), (IV) and (V) are Cu phosphates or pyrophosphates, while non-phosphorous-containing precursors (VI) and (VII), result in mainly CuO. The use of chitosan as a solid-state template/stabilizer induces the formation of CuO and Cu₂O nanoparticles. Copper pyrophosphate (Cu₂P₂O₇) deposited on Si using (IV) as the precursor exhibits single-crystal dots of average diameter 100 nm and heights equivalent to twice the unit cell b-axis (1.5-1.7 nm) and an areal density of 5.1-7.7 Gigadots/in.². Cu₂P₂O₇ deposited from precursor (VI) exhibits unique labyrinthine high surface area deposits. The morphology of CuO deposited on Si from pyrolysis of (VI) depends on the polymer/Cu meta ratio. Magnetic measurements performed using SQUID on CuO nanoparticle networks suggest superparamagnetic behavior. The results give insights into compositional, shape and morphological control of the as-formed nanostructures through the structure of the precursors.     

Increased levels of oxidative stress biomarkers in metal oxides nanomaterial-handling workers

This study assessed oxidatively damaged DNA and antioxidant enzyme activity in workers occupational exposure to metal oxides nanomaterials. Exposure to TiO₂, SiO₂, and ITO resulted in significant lower antioxidant enzymes (glutathione peroxidase and superoxide dismutase) and higher oxidative biomarkers 8-hydroxydeoxyguanosine (8-oxodG) than comparison workers. Statistically significant correlations were noted between plasma and urine 8-oxodG, between white blood cells (WBC) and urine 8-oxodG, and between WBC and plasma 8-oxodG. In addition, there were significant negative correlations between WBC 8-oxodG and SOD and between urinary 8-oxodG and GPx levels. The results showed that urinary 8-oxodG may be considered to be better biomarker.     

Field applications of a bio-trickling filter for the removal of nitrogen oxides from flue gas

A bio-trickling filter (BTF) packed with polyhedral spheres was used to remove nitrogen oxides (NOx) from the flue gas of a coal-fired power plant. The BTF system consistently removed 64–95% of the NOx after start-up and acclimation under dynamic conditions (e.g., 120–240 m³/h flue gas flow rate and inlet 300–900 mg NOx/m³). Scanning electron microscopy of the biofilms that were formed showed a shift in the predominating bacteria. Analyses by PCR-denaturing gradient gel electrophoresis showed that the naturally-selected mixed cultures in the biofilm under a flue gas environment were mainly Klebsiella sp. and Pseudomonas sp.