Variations and evolution of polyubiquitin genes from ciliates

Polyubiquitin genes from seven ciliate species were amplified, cloned and sequenced. It is estimated that Strombidium sulcatum, Euplotes vannus, E. rariseta and Anteholosticha manca have a polyubiquitin gene of 3 repeats, and A. parawarreni, Paramecium caudatum and Pseudokeronopsis flava 4 repeats. The newly obtained ubiquitins mostly differ from that of humans by 1–5 residues in amino acid sequences. A neighbor-joining tree constructed based on monomeric ubiquitin genes supports the monophyly of an assemblage comprising the litostomateans and some oligohymenophoreans, but not the class Spirotrichea. The monomers from the same species are generally placed together and highly supported for the class Litostomatea, the genera Paramecium and Ichthyophthirius, but not for other species. The non-synonymous/synonymous rate ratio (dN/dS) at the protein level are less than 1, and the synonymous nucleotide differences per synonymous site (p_S) from intraspecific comparisons are fairly high (0.02–0.72). These results indicate that ciliates have not only the conserved, but also some quite divergent, polyubiquitin genes and confirm that the polyubiquitin genes in ciliates evolve according to the birth-and-death mode of evolution under strong purifying selection.     

Design and synthesis of a reagent for solid-phase incorporation of the phosphothreonine mimetic (2S,3R)-2-amino-3-methyl-4-phosphonobutyric acid (Pmab) into peptides in a bio-reversible phosphonyl-bis-pivaloyloxymethyl (POM) prodrug form

Reported herein are the synthesis and solid-phase peptide incorporation of N-Fmoc-(2S,3R)-2-amino-3-methyl-4-phosphonobutyric acid bis-pivaloyloxymethyl phosphoryl ester [Fmoc-Pmab(POM)₂-OH, 2] as a phosphatase-stable phosphothreonine (pThr) mimetic bearing orthogonal protection suitable for the synthesis of Pmab-containing peptides having bio-reversible protection of the phosphonic acid moiety. This represents the first report of a bio-reversibly protected pThr mimetic in a form suitable for facile solid-phase peptide synthesis.     

Chiral discrimination and interaction mechanism between enantiomers and serum albumins

The chiral discrimination studies of biological system are theoretically and practically significant for the development of chiral drugs and life science. Our work has embarked upon the interaction between serum albumin (SA) (including human SA and bovine SA), R,S‐1‐(4‐methoxyphenyl)ethylamine, and R,S‐1‐(3‐methoxyphenyl)ethylamine. The formation of intermediate transition state, binding sites, and chiral discrimination ability can be investigated by ultraviolet‐visible spectra and fluorescence spectra. Moreover, both the changes of hydrophobic microenvironment and energy transfer can be detected by synchronous fluorescence spectra and fluorescence lifetime. Copyright © 2013 John Wiley & Sons, Ltd.     

An NAD(P)H Model Reaction: Reduction of α-Keto Esters by the Hantzsch Ester Accelerated by Zinc Complex in the Reformatsky Mixture

Aiming to get useful steroidal alkaloids by tissue culture of Solanum laciniatum Ait., indefinitely growing callus tissue was prepared from the mother plant. Some nutritional requirements for the growth of the callus tissue were studied. By examining steroidal compounds in callus culture, cholesterol, stigmasterol, β-sitosterol, lanosterol, squalene, diosgenin and a new steroidal alkaloid were found to be formed in the callus culture. The new steroidal alkaloid was found to be solasodine derivative containing rhamnose and other unidentified sugars.     

Lipids of Streptomyces sioyaensis

Lipids were extracted with chloroform-methanol from Streptomyces sioyaensis and fractionated on a silicic acid column. Lipids of Streptomyces sioyaensis were mainly composed of neutral lipids, cardiolipin, phosphatidylethanolamines, phosphatidylinositolmonomanno- side and a new lysine-containing lipid.     

Purification and Properties of Sulfite Reductase from Desulfovibrio vulgaris,Miyazaki F

The sulfite reductase of Desulfovibrio vulgaris, strain Miyazaki F (MF), was purified by ammonium sulfate precipitation and chromatography on DEAE-cellulose, Ultrogel AcA34, and hydroxylapatite. The molecular weight was estimated to be 180,000 by gel filtration. It had a subunit structure of α₂β₂; the molecular weight of the α subunit was 50,000 and that of β, 39,000. The absorption spectrum with characteristic peaks at 629 and 409 nm and the amino acid composition resembled those of the sulfite reductase from D. vulgaris, Miyazaki K. The MF enzyme reduced sulfite to trithionate, thiosulfate, and sulfide by hydrogen when coupled with a hydrogenase-methyl viologen system, like other sulfite reductases from Desulfovibrio.     

Ferredoxin-sulfite Reductase from Spinach

Ferredoxin-sulfite reductase (Fd-SiR) [hydrogen-sulfide: ferredoxin oxidoreductase, EC 1.8.7.1] from spinach leaves has been purified to homogeneity by a new procedure. Subunit analysis by sodium dodecyl sulfate gel electrophoresis yielded a single protein band with a molecular weight of 71,000. Gel electrophoresis in non-denaturing media at different acrylamide concentrations gave a molecular weight of 270,000, suggesting that the native enzyme was composed of four identical subunits. In the presence of 0.2 m sodium chloride, however, gel filtration produced a value of 136,000, indicating the presence of dimer in this ionic environment. A plot of substrate (sulfite) concentration versus enzymatic (Fd-SiR) activity yielded a sigmoidal curve, giving a Hill coefficient (n?) of 2.1. Purified Fd-SiR, in the oxidized form, had absorption maxima at 279, 385, 588 and 714nm. Thus the enzyme has the property of a siroheme-containing protein.     

Isolation of Triterpenoid Glycosides (Saikosaponins) from Bupleurum polyclonum Y. Li et S. L. Pan and Their Chemical Structures

Sixteen triterpenoid glycosides, named S13 to S25, S37, S38 and S40, were isolated from the root of Bupleurum polyclonum Y. Li et S. L. Pan, and their structures were determined from NMR spectral analyses. Among them, S24, S37 and S38 were found to be new substances, their structures being established as 30-β-d-glucopyranosyl 30-hydroxysaikosaponin-b₂, 2″-O-acetylsaikosaponin-b₂ and 3″-O>-acetylsaikosaponin-b₂, respectively.     

Ferredoxin-dependent Sulfite Reductase from Spinach Leaves

The intermediate of the aromatization of 4-oxocyclohexanecar-boxylic acid (OHA) to 4-hydroxybenzoic acid (HA) by Coryne-bacterium cyclohexanicum was identified as (+)-4-oxocyclohex-2-enecarboxylic acid (O2A) using a combined system of gas-liquid chromatography (GLC) and a mass spectrometer and polarimeter.