A quick isolation method for mutants with high lipid yield in oleaginous yeast

A novel method has been developed to easily isolate the mutants with high lipid yield after irradiating oleaginous yeast cells with carbon ions of energy of 80 MeV/u. Pre-selection of the mutants after ion irradiation was performed with culture medium in which the concentration of cerulenin, a potent inhibitor of fatty acid synthetase, was at 8.96 mu mol/l. Afterwards, lipid concentration in the fermentation broth of the pre-selected colonies was estimated by the sulfo-phospho-vanillin reaction instead of the conventional methanol-chloroform extraction. Two mutants with high lipid yield have been successfully selected out by the combined method. This easy and simple method is much less time-consuming but very efficient in the mutant isolation, and it has demonstrated great potential on mutation breeding in oleaginous microorganism.

Isolation and extraction of total flavonoids from Epimedium Koreanum Nakai by supercritical fluid extraction

In the present paper is reported the method for the isolation and extraction of total flavonoids of Epimedium Koreanum Nakai by means of supercritical fluid extraction (SFE). By examining pressure, temperature. amounts of modifier and extraction time, the optimized condition of SFE is confirmed as 30 MPa and 60 degreesC. with 70% ethanol as the modifier. The samples were statically extracted for 30 min, followed by dynamic extraction for 120 min at a flow rate of 6 mL/min. The quantitative analysis of total flavonoids was performed by UV-Vis spectrophotometry. Compared with the conventional method, the SFE method is more efficient. more rapid and more friendly environmentally.

Isolation, spectra, structure, and ring-opening polymerization of strained macrocyclic aryl(ether ketone) dimer

Macrocyclic arylene ether ketone dimer was isolated from a mixture of cyclic oligomers obtained by the nucleophilic substitution reaction of bisphenol A and 4,4'-difluorobenzophenone and easily polymerized to high molecular weight linear poly(ether ketone). The cyclic compound was characterized by FTIR, H-1- and C-13-NMR, and single-crystal x-ray diffraction. Analysis of the spectral and crystal structure reveals extreme distortions of he phenyl rings attached to the isopropylidene center and of the turning points of the molecular polygons. The release of the ring strain on ring-opening combined with entropical difference between the linear polymer chain and the more rigid macrocycle at temperatures of polymerization may be the proposed motivating factors in the polymerization of this precursor to high molecular weight poly(ether ketone). (C) 1997 John Wiley & Sons, Inc.

Isolation and X-ray crystallographic structure of cyclic tetramer based on bisphenol A and o-phthaloyl dichloride

The polycrystalline powder of the cyclic tetramer based on bisphenol A and o-phthaloyldichloride has been prepared by recrystallization from nitrobenzene and its crystal structure has been determined by Wide-Angle X-ray Diffraction (WAXD). The unit cell is orthorhombic and its dimensions a = 0.967 6 nm, b = 0.869 9 nm, c = 2.085 9 nm, Space group belongs to Pmmm, With two tetramers per unit cell,the crystal density is 1.36 g . cm(-3), Indices of crystal diffraction peaks are also detailed in the present work.

THE ISOLATION, CHARACTERIZATION AND OXIDATION REACTION OF ACTIVE INTERMEDIATES OF CYTOCHROME-P-450 MODEL SYSTEM OXOCHROMIUM(V) TETRAPHENYLPORPHYRIN COMPLEXES

Oxochromium (V) tetraphenylporphyrin complexes, O = Cr (V) TPP (Cl) PhI. O = Cr-(V) TPP (N3) PhI and O = Cr (V)TPP (p-CH3OC6H4O)1/2PhI were isolated from the reaction of Cr (III) TPP (Cl). Cr (III) TPP (N3) Py or Cr (III) TPP (p-CH3OC6H4O) THF with iodosy

ISOLATION AND PROPERTIES OF MOLYBDENUM SERIES OF HETEROPOLY BLUE WITH KEGGIN STRUCTURE

The present paper reports the methods for preparing and isolating 8 kinds of 1:12 molybdenum series of heteropoly blue complexes KyHzXMo12O40 . nH2O (X=Si, P, As, Ge). The products were characterized by elemental analyses, potential titration, polarograms, cyclic voltammetry, IR spectra, visible-UV spectra, X-ray powder diffraction, XPS and P-31 NMR. The single crystal structure of 4-electron molybdenum-silicon heteropoly blue was measured and the positions of reduced molybdenum atoms were determined, i.e. they were located at Mo(3), Mo(7), Mo(8) and Mo(10). The experimental results show that the heteropoly blue remains Keggin structure. ESR spectra of heteropoly blue solids were first studied, from which it was found that the delocalization extent of 2-electron heteropoly blue and 4-electron heteropoly blue is smaller than that of 1-electron heteropoly blue. The study of thermal properties shows that the thermal stability increases with the increase of the reduction extent of heteropoly blue. The study of redox properties shows that the oxidizing power order of heteropoly blue changes in different mediums, and the polarographic half-wave voltage is found to be dependent on the electronegativity of the hetero atom linearly. It is found that the phosphorus heteropoly blue and arsenic heteropoly blue show a strong anti-acid property.

Isolation and characterization of microsatellites in yellow perch (Perca flavescens)

A total of 45 microsatellite loci from yellow perch, Perca flavescens, were isolated and characterized. Among the 45 microsatellite loci, 32 had more than two alleles. A wild population of P. flavescens (n = 48) was used to examine the allele range of the microsatellite loci. Mendelian inheritance of alleles was confirmed by examining the amplified products in pair-mated families. The number of alleles for the 32 polymorphic loci varied from two to 16, and observed heterozygosity ranged between 0.024 (YP79) and 0.979 (YP60). Cross-species polymorphic amplification in four other Percidae species was successful for 22 loci.

Chinikomycins A and B: Isolation, structure elucidation, and biological activity of novel antibiotics from a marine Streptomyces sp isolate M045

In our screening of marine Streptomycetes for bioactive principles, two novel antitumor antibiotics designated as chinikomycins A (2a) and B (2b) were isolated together with manumycin A (1), and their structures were elucidated by a detailed interpretation of their spectra. Chinikomycins A (2a) and B (2b) are chlorine-containing aromatized manumycin derivatives of the type 64-pABA-2 with an unusual para orientation of the side chains. They exhibited antitumor activity against different human cancer cell lines, but were inactive in antiviral, antimicrobial, and phytotoxicity tests.

Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus

The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degreesC. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degreesC. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N-terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degreesC, with a half-life of about 10 h at 80 degreesC. The activity shows a linear Arrhenius plot at 50-85 degreesC with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90degrees) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant.

ISOLATION AND CHARACTERIZATION OF CHLORELLA SOROKINIANA GXNN01 (CHLOROPHYTA) WITH THE PROPERTIES OF HETEROTROPHIC AND MICROAEROBIC GROWTH

Heterotrophic and anaerobic microalgae are of significance in both basic research and industrial application. A microalga strain was isolated from a wastewater treatment pond and identified as Chlorella sorokiniana Shihira et W. R. Krauss GXNN01 in terms of morphology, physiology, and phylogeny. The strain grows rapidly in heterotrophic or mixotrophic conditions with addition of various carbon sources, and even in anaerobic conditions. The maximum growth rate reached 0.28 d(-1) when using D,L-malate as the carbon source, and the protein content of the microalgae was 75.32% in cell dry weight. The strain was shown to be capable of (1) utilizing D, L-malate only with light, (2) inhibiting photosynthesis in mixotrophic growth, and (3) growing in anaerobic conditions with regular photosynthesis and producing oxygen internally. This study demonstrates the influence of oxygen (aerobic vs. anaerobic) and metabolic regime (autotrophy, mixotrophy, heterotrophy) on the physiological state of the cell.