Proton transport coupled ATP synthesis by the purified yeast H-ATP synthase in proteoliposomes

The H⁺/ATP synthase from yeast mitochondria, MF₀F₁, was purified and reconstituted into liposomes prepared from phosphatidylcholine and phosphatidic acid. Analysis by mass spectrometry revealed the presence of all subunits of the yeast enzyme with the exception of the K-subunit. The MF₀F₁ liposomes were energized by acid-base transitions (ΔpH) and a K⁺/valinomycin diffusion potential (Δφ). ATP synthesis was completely abolished by the addition of uncouplers as well as by the inhibitor oligomycin. The rate of ATP synthesis was optimized as a function of various parameters and reached a maximum value (turnover number) of 120 s^− 1 at a transmembrane pH difference of 3.2 units (at pH_in = 4.8 and pH_out = 8.0) and a Δφ of 133 mV (Nernst potential). Functional studies showed that the monomeric MF₀F₁ was fully active in ATP synthesis. The turnover increased in a sigmoidal way with increasing internal and decreasing external proton concentration. The dependence of the turnover on the phosphate concentration and the dependence of K_M on pH_out indicated that the substrate for ATP synthesis is the monoanionic phosphate species H₂PO₄⁻.     

Statistical screening and optimization of process variables for xylanase production utilizing alkali-pretreated rice husk

With the objective of the production of xylanase, local raw material (rice husk) and the indigenous isolate, Aspergillus niger ITCC 7678, were studied. Optimization of the cultivation system for enhancing xylanase production was studied via submerged fermentation. Statistical procedures were employed to study the effect of process variables, such as alkali-pretreated rice husk (as carbon source), NaNO₃ (as nitrogen source), KH₂PO₄, KCl, Tween 80 (as surfactant), MgSO₄, FeSO₄·7H₂O, pH, particle size, agitation, and temperature, on xylanase production by A. niger. The effect and significance of the variables was studied using Plackett–Burman (PBD) and central composite statistical design (CCD). It was found that alkali pretreated rice husk (weight/volume), pH, temperature, and NaNO₃ significantly influence xylanase production. So, these four factors were further optimized by CCD, and it was found that maximum xylanase activity of 10.9 IU/ml was observed at (6.5 % w/v) rice husk, pH (5.5), temperature (32.5 °C), and NaNO₃ (0.35 % w/v) concentration. Under optimum conditions, xylanase production was also studied at the bioreactor level and showed 12.8 % enhanced xylanase activity.     

Analysis of peroxidase activity of rice (Oryza sativa) recombinant hemoglobin 1: Implications for in vivo function of hexacoordinate non-symbiotic hemoglobins in plants

In plants, it has been proposed that hexacoordinate (class 1) non-symbiotic Hbs (nsHb-1) function in vivo as peroxidases. However, little is known about peroxidase activity of nsHb-1. We evaluated the peroxidase activity of rice recombinant Hb1 (a nsHb-1) by using the guaiacol/H₂O₂ system at pH 6.0 and compared it to that from horseradish peroxidase (HRP). Results showed that the affinity of rice Hb1 for H₂O₂ was 86-times lower than that of HRP (K_m = 23.3 and 0.27 mM, respectively) and that the catalytic efficiency of rice Hb1 for the oxidation of guaiacol using H₂O₂ as electron donor was 2838-times lower than that of HRP (k_cat/K_m = 15.8 and 44 833 mM⁻¹ min⁻¹, respectively). Also, results from this work showed that rice Hb1 is not chemically modified and binds CO after incubation with high H₂O₂ concentration, and that it poorly protects recombinant Escherichia coli from H₂O₂ stress. These observations indicate that rice Hb1 inefficiently scavenges H₂O₂ as compared to a typical plant peroxidase, thus indicating that non-symbiotic Hbs are unlikely to function as peroxidases in planta.     

Release of monomeric sugars from Miscanthus sinensis by microwave-assisted ammonia and phosphoric acid treatments

Microwave-assisted ammonium hydroxide (NH₄OH) followed by phosphoric acid (H₃PO₄) treatments were used to release monomeric sugars from Miscanthus sinensis grown in Cha-Chueng-Sao province, Thailand. Treatment with 1.0% (w/v) NH₄OH, 15:1 liquid-to-solid ratio (LSR) at 120 °C temperature for 15 min liberated 2.9 g of monomeric sugars per 100 g of dried biomass, whereas the corresponding yield for a treatment with 1.78% v/v H₃PO₄, 15:1 LSR at 140 °C for 30 min was 62.3 g/100 g. The two-stage pretreatment, treatment with NH₄OH at 120 °C temperature for 15 min followed by treatment with H₃PO₄ at 140 °C for 30 min, impressively provided the highest total monomeric sugar yield of 71.6 g/100 g dried biomass.     

Isolation of cellulose-hydrolytic bacteria and applications of the cellulolytic enzymes for cellulosic biohydrogen production

Nine cellulolytic bacterial strains were isolated from soil sample taken in southern Taiwan. Through 16S rRNA sequence matching; eight of those isolates belong to Cellulomonas sp., while the other one belongs to Cellulosimicrobium cellulans. The activity of cellulolytic enzymes (cellulases and xylanase) produced from those strains was mainly present extracellularly and the enzyme production was dependent on cellulosic substrates (xylan, rice husk and rice straw) used for growth. HPLC analysis confirmed the bacterial hydrolysis of these cellulosic substrates for soluble sugars production. The efficiency of fermentative H₂ production from the enzymatically hydrolyzed rice husk was examined with seven H₂-producing pure bacterial isolates. With an initial reducing sugar concentration of 0.36 g l⁻¹, only Clostridium butyricum CGS5 exhibited efficient H₂ production from the rice husk hydrolysates with a cumulative H₂ production and H₂ yield of 88.1 ml l⁻¹ and 19.15 mmol H₂ (g reducing sugar)⁻¹ (or 17.24 mmol H₂ (g cellulose)⁻¹), respectively.     

A stable second-order NLO and luminescent Cd(II) complex based on 6-hydroxynicotinic acid

Employing an unsymmetrical 6-hydroxynicotinic acid as a bridging ligand, we have prepared a new NLO-active two-dimensional cadmium coordination polymer [Cd₈(nicO)₈(phen)₈ · (H₂O)]_n (1) (H₂nicO = 6-hydroxynicotinic acid, phen = 1,10-phenanthroline) under hydrothermal condition. At room temperature, 1 shows strong fluorescent emission bands in the solid state and a SHG efficiency of about 8 times that of KDP (KH₂PO₄). On the basis of TG/DTA analyses, the structure is thermally stable up to 370 °C.     

Synthesis, crystal structure and luminescent behaviour of coordination complexes of copper with bi- and tridentate amines and phosphonic acids

The synthesis and crystal structure of four new copper(I) and copper(II) supramolecular amine, and amine phosphonate, complexes is reported. Reaction of copper(I) with 2-,9-dimethyl-1-10-phenanthroline (dmp) produced a stable 4-coordinate Cu(I) species, [Cu^(I)(dmp)₂]Cl · MeOH · 5H₂O (2), i.e., the increased steric hindrance in the ‘bite’ area of dmp did not prevent interaction with the metal and provided protection against oxidation which was not possible for the phen analogue [R. Clarke, K. Latham, C. Rix, M. Hobday, J. White, CrystEngCommun. 7(3) (2005), 28-36]. Subsequent addition of phenylphosphonic acid to (2) produced two structures from alternative synthetic routes. An ‘in situ’ process yielded red block Cu(I) crystals, [Cu^(I)(dmp)₂] · [C₆H₅PO₃H₂ · C₆H₅PO₃H] (4), whilst recrystallisation of (2) prior to addition of the acid (‘stepwise’ process) produced a green, needle-like Cu(II) complex, [Cu^(II)(dmp) · (H₂O)₂ · C₆H₅PO₂(OH)] [C₆H₅PO₂(OH)] (3). However, addition of excess dmp during the ‘stepwise’ process forced the equilibrium towards product (4) and resulted in an optimum yield (99%). The structure of (4) was similar to the phen analogue, [Cu^(II)Cl(phen)₂] · [C₆H₅PO₂(OH) · C₆H₅PO(OH)₂] (1) [R. Clarke, K. Latham, C. Rix, M. Hobday, J. White, CrystEngCommun. 7(3) (2005), 28-36], but the presence of dmp exerted some influence on global packing, whilst (3) exists as a polymeric layered material. In contrast, reaction of copper(I) with di-2-pyridyl ketone (dpk), followed by phenylphosphonic acid produced purple/blue Cu(II) species, [Cu^(II)(dpk · H₂O)₂] Cl₂ · 4H₂O (5), and [Cu^(II)(dpk · H₂O)₂] · [C₆H₅PO₂(OH)₂ · C₆H₅PO(OH)₂] (6), respectively, i.e., in both cases oxidation of copper occurred. Solid-state luminescence was observed in (2) and (4). The latter showing a 5-fold enhancement in intensity.     

Preparation, structure, solid state and gas phase stability of the mixed neodymium nitrate-chloride complex NdCl(NO3)2{[(MeO)2PO]2C(OH)Bu}2

The preparation and structure of the mixed anion complex NdCl(NO₃)₂{[(MeO)₂PO]₂C(OH)^tBu}₂ are reported. Single crystal X-ray diffraction shows that the bisphosphonate is bonded via both phosphoryl groups and the nitrates act as bidentate ligands. Intramolecular H-bonding is seen between the OH and the coordinated nitrate and chloride ligands. Thermal decomposition in the solid state is by loss of methyl nitrate. Electrospray mass spectrometry shows that loss of chloride is preferred over loss of nitrate in the gas phase. Attempted preparation of NdCl₂(NO₃){[(MeO)₂PO]₂C(OH)^tBu}₂ leads to the formation of a product approximating to [Nd{^tBu(OH)C(PO₃H₂)₂}₂]₂H · NO₃ · (PO₄H₂)₂. Electrospray mass spectrometry and elemental analysis confirm the presence of the [^tBu(OH)C(PO₄H₂)₂]⁻ in the decomposition products.     

Recovery of antibiotics by aqueous two-phase partition —Partitioning behavior of pure acetylspiramycin solution in polyethylene glycol/potassium phosphate aqueous two-phase systems

Summary The effects of average molecular weight of PEG, concentrations of PEG and KH₂PO₄ and pH on the partition equilibrium of acetylspiramycin in PEG/KH₂PO₄ aqueous two-phase systems were studied in detail. The partition coefficients of acetylspiramycin in PEG/ KH₂PO₄ systems were measured at room temperature 25 °C. It was found that acetylspiramycin partitioned unevenly in the aqueous two-phase systems composed of PEG and KH₂PO₄ and could be purified by this technique. A suitable phase-forming system (pH=6.7, 12w/w% PEG2000, 11w/w% KH₂PO₄) was found out after partition coefficient (Kp=42) , extraction ratio (=96%) and recovery ratio(R=98.8%) were investigated comprehensively in this paper.Hua qiang is one of the cooperators of the experimetal.     

Alkaline peroxide assisted wet air oxidation pretreatment approach to enhance enzymatic convertibility of rice husk

Pretreatment of rice husk by alkaline peroxide assisted wet air oxidation (APAWAO) approach was investigated with the aim to enhance the enzymatic convertibility of cellulose in pretreated rice husk. Rice husk was presoaked overnight in 1% (w/v) H₂O₂ solution (pH adjusted to 11.5 using NaOH) (equivalent to 16.67 g H₂O₂ and 3.63 g NaOH per 100 g dry, untreated rice husk) at room temperature, followed by wet air oxidation (WAO). APAWAO pretreatment resulted in solubilization of 67 wt % of hemicellulose and 88 wt % of lignin initially present in raw rice husk. Some amount of oligomeric glucose (?8.3 g/L) was also observed in the APAWAO liquid fraction. APAWAO pretreatment resulted in 13‐fold increase in the amount of glucose that could be obtained from otherwise untreated rice husk. Up to 86 wt % of cellulose in the pretreated rice husk (solid fraction) could be converted into glucose within 24 hours, yielding over 21 g glucose per 100 g original rice husk. Scanning electron microscopy was performed to visualize changes in biomass structure following the APAWAO pretreatment. Enzymatic cellulose convertibility of the pretreated slurry at high dry matter loadings was also investigated. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011     

Solid state coordination chemistry of organodiphosphonates with copper(II) and auxilliary aromatic nitrogen heterocyclic ligands

The hydrothermal reactions of CuSO₄ · 5H₂O, a phosphonate ligand and an appropriate aromatic nitrogen heterocycle yield a series of materials of the Cu(II)/phosphonate/nitrogen ligand donor family. Two of the materials [Cu(2,2′-bpy)(HO₃PCH₂CH₂CH₂PO₃H)] (1) and [Cu(4,4′-bpy)(H₂O)₂(HO₃PCH₂CH₂CH₂CH₂PO₃H)] (2) are two-dimensional, while [{Cu(2,2′-bpy)}₂(O₃PC₆H₄PO₃)] · 8H₂O (3 · 8H₂O) is three-dimensional. When 1,3,5-benzenetricarboxylic acid is introduced as a reactant, the one-dimensional material [Cu(2,2′-bpy){OC₆H₂(CO₂H)₃}(HOPC₆H₅)] (4) is isolated. This is an example of an in situ hydrothermal ligand transformation in which the 1,3,5-benzene tricarboxylic acid is hydroxylated to give 1,3,5-benzene tricarboxylic acid-2-hydroxide. Compound 5 [Cu(terpy)(HO₃PC₆H₄PO₃H] is molecular.     

Hydrothermal synthesis, crystal structure and physical properties of a new gadolinium phosphite hydrate

A new compound Gd₄(H₂O)₅(HPO₃)₆ (1) was isolated from the reaction of gadolinium chloride GdCl₃ · nH₂O with H₃PO₃ under hydrothermal conditions. The crystal structure was solved by means of single-crystal X-ray diffraction. The structure is composed of parallel gadolinium-oxygen polyhedra chains connected by phosphite ions, and shows three-dimensional (3D) open-framework with eight-membered ring (8MR) channels. The structure is different from its counterpart Eu₂(H₂O)_2.5(PO₃H)₃ due to the effect of lanthanide contraction. The intensive intrinsic UV emission of this compound at λ_max = 312 nm comes from the spin-forbidden ⁶P_7/2 → ⁸S_7/2 f-f transition of the Gd³⁺ ions. No magnetic order was observed for this compound.     

Novel oxorhenium(V) ‘3+1’ mixed ligand complexes with 3-thiapentane-1,5-dithiolate and functional mercaptobenzoyl amino acid ethyl ester

Oxorhenium(V) complexes with ‘3+1’ mixed ligands, [ReO(SSS)L], where SSS is η³-(SCH₂CH₂SCH₂CH₂S), L = η¹-(C₆H₄COOH-4-S), η¹-(C₆H₄CONHCH₂COOEt-4-S), η¹-(C₆H₄CONHCH(CH₃)COOEt-4-S), and η¹-(C₆H₄CONHCH(CH₂Ph)COOEt-4-S), have been synthesized. These L ligands and [ReO(SSS)L] complexes were characterized by IR, ¹H NMR, ¹³C NMR, and MAS spectrometers. Molecular structure of [ReO(SSS){η¹-(C₆H₄COOH-4-S)}] complex was determined to be a distorted square pyramidal by single crystal X-ray analytical method.     

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.     

Transport of DMAA and MMAA into rice (Oryza sativa L.) roots

Arsenate (As(V)) transport into plant cells has been well studied. A study on rice (Oryza sativa L.) showed that arsenite is transported across the plasma membrane via glycerol transporting channels. Previous studies reported that the dimethylarsinic acid (DMAA) and monomethylarsonic acid (MMAA) uptake in duckweed (Spirodela polyrhiza L.) differed from that of As(V), and was unaffected by phosphate (H₂PO₄). This article reports the transport mechanisms of DMAA and MMAA in rice roots. Linear regression analysis showed that the DMAA and MMAA uptake in rice roots increased significantly (p ≤ 0.0002 and ≤0.0001 for DMAA and MMAA, respectively) with the increase of exposure time. Concentration-dependent influx of DMAA and MMAA showed that the uptake data were well described by Michaelis-Menten kinetics. The MMAA influx was higher than that of DMAA. The DMAA and MMAA uptake in rice roots were decreased significantly (p ≤ 0.0001 and ≤0.0077 for DMAA and MMAA, respectively) with the increase of glycerol concentration indicating that DMAA and MMAA were transported into rice roots using the same mechanisms of glycerol. Glycerol is transported into plant cells by aquaporins, and DMAA and MMAA are transported in a dose-dependent manner of glycerol which reveals that DMAA and MMAA are transported into rice roots through glycerol transporting channels. The DMAA and MMAA concentration in the solution did not affect the inhibition of their uptake rate by glycerol.     

Use of glycerol for producing 1,3-dihydroxyacetone by Gluconobacter oxydans in an airlift bioreactor

1,3-Dihydroxyacetone can be produced by biotransformation of glycerol with glycerol dehydrogenase from Gluconobacter oxydans cells. Firstly, improvement the activity of glycerol dehydrogenase was carried out by medium optimization. The optimal medium for cell cultivation was composed of 5.6 g/l yeast extract, 4.7 g/l glycerol, 42.1 g/l mannitol, 0.5 g/l K₂HPO₄, 0.5 g/l KH₂PO₄, 0.1 g/l MgSO₄·7H₂O, and 2.0 g/l CaCO₃ with the initial pH of 4.9. Secondly, an internal loop airlift bioreactor was applied for DHA production from glycerol by resting cells of G. oxydans ZJB09113. Furthermore, the effects of pH, aeration rate and cell content on DHA production and glycerol feeding strategy were investigated. 156.3 ± 7.8 g/l of maximal DHA concentration with 89.8 ± 2.4% of conversion rate of glycerol to DHA was achieved after 72 h of biotransformation using 10 g/l resting cells at 30 °C, pH 5.0 and 1.5 vvm of aeration rate.     

A high-pressure, Raman spectroscopic study of bis(pyrophosphato)tetrazinc(II) and -tetramanganese(II) decahydrate salts

The Raman spectra of crystalline bis(pyrophosphato)tetrazinc(II) and -manganese(II) decahydrates, M₄(P₂O₇)₂ · 10H₂O (M = Zn and Mn), have been investigated at room temperature. The Mn(II) salt readily underwent photodecomposition upon extended exposure to 785-nm laser irradiation and so could not be examined under high pressures in a diamond-anvil cell. The Zn(II) salt was much more stable and was studied successfully at pressures up to ∼35 kbar. The pressure dependences of the δ_sPO₂, ν_sPOP, ν_asPOP, ν_sPO₂, and ν_asPO₂ internal modes of the pyrophosphate group and of an external H₂O mode indicated the occurrence of a pressure-induced phase transition in the 15-20 kbar region. Further confirmation of this structural change came from the appearance of four initially forbidden modes in the 19-26 kbar range. This phase transition proved to be reversible upon release of the applied pressure. The pressure dependences before and after the phase transition were −0.22 to 0.66 and 0.06 to 0.33 cm⁻¹/kbar, respectively.     

Effects of key operational parameters on biohydrogen production via anaerobic fermentation in a sequencing batch reactor

In this study, a series of tests were conducted in a 6 L anaerobic sequencing batch reactor (ASBR) to investigate the effect of pH, hydraulic retention time (HRT) and organic loading rate on biohydrogen production at 28 °C. Sucrose was used as the main substrate to mimic carbohydrate-rich wastewater and inoculum was prepared from anaerobic digested sludge without pretreatment. The reactor was operated initially with nitrogen sparging to form anaerobic condition. Results showed that methanogens were effectively suppressed. The optimum pH value would vary depending on the HRT. Maximum hydrogen production rate and yield of 3.04 L H₂/L reactor d and 2.16 mol H₂/mol hexose respectively were achieved at pH 4.5, HRT 30 h, and OLR 11.0 kg/m³ d. Two relationships involving the propionic acid/acetic acid ratio and ethanol/acetic acid ratio were derived from the analysis of the metabolites of fermentation. Ethanol/acetic acid ratio of 1.25 was found to be a threshold value for higher hydrogen production.     

Ln2(O2CCH2PO3)2(H2O)3 · H2O (Ln = La, Pr, Nd, Sm): Layered lanthanide phosphonoacetates containing two-dimensional -Ln-O- linkages

Four novel isostructural lanthanide phosphonate compounds with formula Ln₂(O₂CCH₂PO₃)₂(H₂O)₃ · H₂O [Ln = La (1), Pr (2), Nd (3), Sm (4)] have been prepared through hydrothermal reactions of phosphonoacetate acid and lanthanide nitrates. All show layered structures made up of {LnO₉} polyhedra and {CPO₃} tetrahedra with the lattice water molecules locating between the layers. Within the layer, chains of edge-sharing {LnO₉} polyhedra are connected via corner-sharing by phosphonate oxygens forming a two-dimensional -Ln-O- linkage. Thermal analyses and XRD measurements reveal that the framework structures can be maintained up to 400 °C.     

Syntheses, characterizations and crystal structures of two lead(II) diphosphonates: Pb2[NH(CH2PO3)2] · 2H2O and Pb3[HO2C(CH2)3NH(CH2PO3)2]2 · 2H2O

Hydrothermal reactions of lead(II) acetate and HO₂C(CH₂)₃N(CH₂PO₃H₂)₂ at 170 and 140 °C, respectively, resulted in two different lead diphosphonates, namely, Pb₂[NH(CH₂PO₃)₂] · 2H₂O (1), in which the butyric acid moiety of the HO₂C(CH₂)₃N(CH₂PO₃H₂)₂ has been cleaved and a novel layered compound, Pb₃[HO₂C(CH₂)₃NH(CH₂PO₃)₂]₂ · 2H₂O (2). Their crystal structures have been determined by single crystal X-ray diffraction. In compound 1, the interconnection of the lead(II) ions by bridging amino-diphosphonate ligands leads to the formation of a 3D network. Compound 2 features an unusual triple-layer structure with the non-coordinated butyric acid moieties as pendant groups between the layers.