Whole-cell biocatalysis: Evaluation of new hydrophobic ionic liquids for efficient asymmetric reduction of prochiral ketones

A biphasic process design is often applied in whole-cell biocatalysis if substrate and product have low water solubility, are unstable in water or toxic for the biocatalyst. Some water immiscible ionic liquids (ILs) with adequate distribution coefficients have already been applied successfully as second liquid phase, which acts as a substrate reservoir and in situ extractant for the product. In this work, 12 new ILs were evaluated with respect to their applicability in biphasic asymmetric reductions of prochiral ketones in comparison to 9 already published ILs. The ILs under study are composed of seven different cations and three different anions. Recombinant Escherichia coli was used as whole-cell biocatalyst overexpressing the genes of a Lactobacillus brevis alcohol dehydrogenase (LB-ADH) and a Candida boidinii formate dehydrogenase (CB-FDH) for cofactor regeneration. Best results were achieved if ionic liquids with [PF6]- and [NTF]-anions were applied, whereas [FAP]-ILs showed minor qualification, e.g., the use of [HMPL][NTF] as second liquid phase for asymmetric synthesis of (R)-2-octanol resulted in a space–time-yield of 180 g L⁻¹ d⁻¹, a chemical yield of 95% and an enantiomeric excess of 99.7% in a simple batch process.     

低碳发展背景下北京市土地利用与生态系统服务多情景模拟研究

在全球气候变化的背景下,合理协调低碳减排与经济效益之间的关系,进而指导土地利用优化配置,对于推动经济社会低碳转型与生态系统服务价值的提升具有重要意义。通过耦合多目标规划(MOP)与斑块生成土地利用模拟模型(PLUS),针对北京市构建了自然演变、经济优先与低碳发展这3种未来发展情景,并对不同情景下2030年土地利用与生态系统服务价值进行模拟评估。在此基础上,进一步对北京市未来生态系统服务提升区域进行识别,将传统的多情景土地利用模拟研究进行了延伸与扩展。结果表明：(1)3种情景下的土地利用结构与空间布局存在差异,低碳发展情景下东南部平原区建设用地的扩张与耕地面积的减少趋势得到了有效控制,西北部山区的林地面积有较大幅度的增长。(2)低碳发展情景下的生态系统服务价值总量约为590亿元,高于另外两种情景,且从空间分布上来看,该情景下服务价值提升区域的面积也显著高于其他情景。(3)低碳发展情景下,北京市生态系统服务提升潜力较大的区域面积占比约7.98%、潜力中等区域的面积占比约26.57%,潜力较低区域的面积占比约65.45%。     

Selective inhibition of methanogenesis to enhance ethanol and n-butyrate production through acetate reduction in mixed culture fermentation

Acetate reduction is an alternative digestion process to convert organic waste into ethanol. Using acetate for fuel ethanol production offers the opportunity to use organic waste materials instead of sugar-containing feedstock. Methanogenesis, however, competes with acetate reduction for acetate and hydrogen and lowers the final efficiency. The aim of this research is to selectively inhibit methanogenesis and to enhance acetate reduction. Acetate reduction was stimulated in batch tests at pH between 4.5 and 8; and at pH 6 with and without thermal pre-treatment. It was found that methanogenesis was selectively inhibited while acetate reduction was enhanced after thermal pre-treatment incubated at pH 6. Initially the acetate reduction yielded 7.7 ± 3.2 mM ethanol with an efficiency of 60.2 ± 8.7%, but later on it was consumed to form 7.02 ± 0.85 mM n-butyrate with an efficiency of 76.2 ± 14.0%. It was the first time demonstrated that n-butyrate can be produced by mixed cultures from only acetate and hydrogen.     

Regioselectivity in the glycosylation of 5-(3-chlorobenzo[b]thien-2-yl)-4H-1,2,4-triazole-3-thiol

The glycosylation of 5-(3-chlorobenzo[b]thien-2-yl)-4H-1,2,4-triazole-3-thiol (1) and its 3-benzylsulfanyl and 3-methylsulfanyl derivatives with different glycosyl halides 2-4 has been studied in presence of base. The S-glycosides 5-7 were obtained in the presence of triethylamine, whereas the respective S,N⁴-bis(glycosyl) derivatives 8-10 were synthesized in the presence of potassium carbonate; the S,N²-bis(glycosyl) isomer 11 could also be isolated in the case of the galactosyl analog. Similarly, after protecting 1 as 3-benzyl(methyl)sulfanyl derivatives 12 or 13, the N⁴-glycosyl analogs 14-19 as well as minor amounts of S,N²-bis(galactosyl) isomers 20 and 21 were formed. The theoretical calculations using AM1 semiempirical methods agreed with the experimental results. Microwave irradiation (MWI) led to higher yields in much less time than the conventional methods, and no change in regioselectivity has been noticed.     

Voltammetry of half-sandwich manganese group complexes of η-PhC3B7H9 and η-C60Bn2PhH2, two ligands that are cyclopentadienyl mimicks

The potentials of a series of one-electron oxidation and reduction reactions have been determined for manganese group half-sandwich complexes of the tricarbadecaboranyl ligand PhC₃B₇H₉ and the penta-organo fullerene ligand C₆₀Bn₂PhH₂ (Bn = benzyl). The anodic processes were studied in CH₂Cl₂ and the cathodic processes were studied in both CH₂Cl₂ and THF, the supporting electrolyte being [NBu₄][B(C₆F₅)₄]. The manganese complex Mn(CO)₂(PMe₃)(PhC₃B₇H₉) (1) is a member of a three-electron transfer series which includes oxidation to 1⁺ (0.51 V versus ferrocene) and successive reductions to 1⁻ (−1.66 V) and 1²⁻ (−1.77 V). Both the oxidation and reduction of the closely-related complex Mn(CO)₂(PPh₃)(PhC₃B₇H₉) (2) are chemically irreversible under slow-scan cyclic voltammetry conditions. The rhenium complex Re(CO)₂(PPh₃)(PhC₃B₇H₉) (3) oxidizes (E_1/2 = 0.82 V versus ferrocene) to a radical cation which, unlike its cyclopentadienyl analogue, shows no evidence of dimerization. Oxidation of the fullerene-based complex Re(CO)₃(C₆₀Bn₂PhH₂) is more facile than that of its cyclopentadienyl analogue, in contrast to previous findings in this class of metal-fullerene derivatives. An electrochemical ligand factor, E_L, of 0.63 is calculated for the PhC₃B₇H₉ ligand in manganese group half-sandwich complexes.     

Transformation of metals and metal ions by hydrogenases from phototrophic bacteria

The ability of hydrogenases isolated from Thiocapsa roseopersicina and Lamprobacter modestohalophilus to reduce metal ions and oxidize metals has been studied. Hydrogenases from both phototrophic bacteria oxidized metallic Fe, Cd, Zn and Ni into their ionic forms with simultaneous evolution of molecular hydrogen. The metal oxidation rate decreased in the series Zn>Fe>Cd>Ni and depended on the pH. The presence of methyl viologen in the reaction system accelerated this process. T. roseopersicina and L. modestohalophilus cells and their hydrogenases reduced Ni(II), Pt(IV), Pd(II) or Ru(III) to their metallic forms under H₂ atmosphere. These results suggest that metals or metal ions can serve as electron donors or acceptors for hydrogenases from phototrophic bacteria.     

Evaluation of a modified selective differential medium for the isolation of Stenotrophomonas maltophilia

VIA medium for Stenotrophomonas maltophilia was modified by substituting meropenem (16 mg/L) for imipenem. S. maltophilia grew from 40% of drains sampled within a hospital and surrounding locations in Perth, Western Australia. The specificity of the new medium for S. maltophilia was 62%, and all contaminating bacteria were easily distinguishable by phenotypic tests and PCR.     

Rhizobium strain effects on pea: The relation between nitrogen accumulation, phosphoenolpyruvate carboxylase activity in nodules and asparagine in root bleeding sap

Pisum sativum L. cv. Bodil was infected with various strains of Rhizobium leguminosarum (R501, 128c53, B155, 18a or 1044). The Rhizobium genotype influenced the activity of the plant enzyme phosphoenoipyruvate (PEP) carboxylase (EC 4.1.1.31), and the assimilation of fixed N in the root nodules. The specific activity of nodule PEP carboxylase was lowest in the symbioses, which accumulated the least total N (R501 and 128c53). The root bleeding sap of the less effective symbioses contained a lower proportion of asparagine and a higher proportion of glutamine than the more effective symbioses (B155,18a and 1044). The N yield of the symbioses was related neither to the net respiratory CO₂ evolution of the root system nor to the nitrogenase linked nodule respiration. The lower yielding symbioses accumulated a larger proportion of the fixed N in the nodules due to a higher proportion of total dry weight contained in the nodule tissue. However, the concentration of soluble protein in the nodules of the lower-yielding symbioses was lower than that recorded for the higher yileding symbioses. The effect of the Rhizobium strains on N yield was maintained at maturity, and reflected in seed yields.     

1,5-Disubstituted indole derivatives as selective human neuronal nitric oxide synthase inhibitors

A series of 1,5-disubstituted indole derivatives was designed, synthesized and evaluated as inhibitors of human nitric oxide synthase. A variety of flexible and restricted basic amine side chain substitutions was explored at the 1-position of the indole ring, while keeping the amidine group fixed at the 5-position. Compounds having N-(1-(2-(1-methylpyrrolidin-2-yl)ethyl)- (12, (R)-12, (S)-12 and 13) and N-(1-(1-methylazepan-4-yl)- side chains (14, 15, (-)-15 and (+)-15) showed increased inhibitory activity for the human nNOS isoform and selectivity over eNOS and iNOS isoforms. The most potent compound of the series for human nNOS (IC(50)=0.02 μM) (S)-12 showed very good selectivity over the eNOS (eNOS/nNOS=96-fold) and iNOS (iNOS/nNOS=850-fold) isoforms.