Purification, characterization, and cDNA cloning of opine dehydrogenases from the polychaete rockworm Marphysa sanguinea.

Alanopine dehydrogenase (AlDH) and three isoforms of strombine/alanopine dehydrogenase (St/AlDH) were purified from muscle tissue of the polychaete rockworm Marphysa sanguinea. The four enzymes, which can be distinguished by the isoelectric point, are monomeric 42 kDa proteins, possess similar pH-activity profiles, and display specificity for pyruvate and NAD(H). The three isoforms of St/AlDH show equivalent Km and Vmax for glycine and L-alanine and for D-strombine and meso-alanopine.

Tauropine dehydrogenase from the marine sponge Halichondria japonica is a homolog of ornithine cyclodeaminase/mu-crystallin.

The partial amino acid sequence including the N- and C-terminal portions of tauropine dehydrogenase (EC 1.5.1.

The amino acid sequence of tauropine dehydrogenase from the polychaete Arabella iricolor.

The amino acid sequence of tauropine dehydrogenase (EC 1.5.1.

Complementary DNA cloning and molecular evolution of opine dehydrogenases in some marine invertebrates.

The complete complementary DNA sequences of genes presumably coding for opine dehydrogenases from Arabella iricolor (sandworm), Haliotis discus hannai (abalone), and Patinopecten yessoensis (scallop) were determined, and partial cDNA sequences were derived for Meretrix lusoria (Japanese hard clam) and Spisula sachalinensis (Sakhalin surf clam). The primers ODH-9F and ODH-11R proved useful for amplifying the sequences for opine dehydrogenases from the 4 mollusk species investigated in this study. The sequence of the sandworm was obtained using primers constructed from the amino acid sequence of tauropine dehydrogenase, the main opine dehydrogenase in A.

Presence of free D-amino acids in microalgae.

Several species of microalgae (phytoplankton), 4 species of freshwater algae and 4 species of marine diatoms, were cultured germ-free in the laboratory. The presence of free D-amino acids was verified in these species by a reversed-phase HPLC analysis. D-Aspartate was detected in all the microalgae examined, but D-alanine was only present in the marine diatoms.

Occurrence of beta-alanine-specific opine dehydrogenase in the muscle of the limpet Cellana grata Gould (Archaeogastropoda).

The muscular tissues of the limpet Cellana grata exhibited beta-alanopine dehydrogenase (beta-AlDH) activity in addition to tauropine dehydrogenase (TaDH) activity and weak lactate dehydrogenase activity. Opine dehydrogenases (OpDHs) were purified, and two different types of OpDH, i.e.

Tauropine dehydrogenase from the starfish Asterina pectinifera (Echinodermata: Asteroidea): presence of opine production pathway in a deuterostome invertebrate.

Tauropine dehydrogenase (tauropine:NAD oxidoreductase; TaDH) was purified to homogeneity from the body wall of the starfish Asterina pectinifera Müller at Troschel(Echinodermata: Asteroidea) by means of (NH4)2SO4 precipitation followed by column chromatographies in DEAE-cellulose, Sephadex G75, Macro-prep ceramic hydroxyapatite, PBE 94, and Toyopearl HW50S. The enzyme was a monomeric protein of approximately 42000 Da and pI 5.2.

Tauropine dehydrogenase from the sandworm Arabella iricolor (Polychaeta: Errantia): purification and characterization.

This is the first report of the purification of tauropine dehydrogenase (NAD: tauropine oxidoreductase) from a polychaete worm. In the sandworm Arabella iricolor Montagu (Polychaeta: Errantia), two forms of TaDH were detected: major component (pl = 7.5) and minor one (pl = 6.

Effects of octopine on the serum cholesterol level in rats.

The effects of dietary octopine, which is one of the major extractive component of marine molluscs, on the level of serum and liver cholesterol of rats fed with cholesterol-enriched or cholesterol-free diets were investigated. Dietary supplementation with 1.5% octopine in a cholesterol-enriched diet significantly decreased the serum total- and VLDL+LDL-cholesterol levels and by contrary increased the serum HDL-cholesterol level in rats.

Specific determination of histamine in fish by high-performance liquid chromatography after diazo coupling.

A new approach to the pre-column derivatization and analysis of histamine by HPLC is described, the method being based upon the formation of a diazo-coupled derivative of histamine. This derivatization method is simple, efficient, sensitive and specific for histamine and other imidazole compounds without a preliminary clean-up step. The liquid chromatographic system allows for rapid, bonded-phase separation with visible-light detection of these compounds within 20 min at a 10-pmol sensitivity.

Nonradioactive assay for adenosine 5'-phosphosulfate sulfotransferase using reversed-phase ion-pair high-performance liquid chromatography.

A nonradioactive and simple method for the assay of adenosine 5'-phosphosulfate sulfotransferase (APSSTase) is described. The method is based upon the quantitative measurement of the substrate adenosine 5'-phosphosulfate and/or product AMP by ion-pair reversed-phase high-performance liquid chromatography on a C18 column. Isocratic elution renders the method simple and fast.

Occurrence of free D-aspartic acid in marine macroalgae.

The biologically rare D-aspartic acid was found in extracts of marine macroalgae. DL-aspartic acid was isolated from the Phaeophyta, Hizikia fusiformis, by ion-exchange chromatography, and crystallized from aqueous ethanol. It was characterized by optical-rotatory-dispersion analysis and reversed-phase HPLC analysis.

Determination of octopine by pre-column fluorescence derivatization using benzoin.

A sensitive fluorimetric method for the determination of octopine, a member of opine family, is presented. The method is based on the formation of a fluorescent derivative of octopine with benzoin and the separation by high performance liquid chromatography using a reversed-phase column (Kaseisorb LC ODS-300) within 20 min. The octopine derivative is completely separated from other guanidino compounds including arginine which is generally very high in marine invertebrates.

Electroporation as a new technique for producing transgenic fish.

A recombinant plasmid, pMV-GH, containing rainbow trout growth hormone cDNA fused to mouse metallothionein I promoter, was introduced into medaka (Oryzias latipes) by electroporation. Of 3109 fertilized eggs treated with electric pulses (750 V/cm, 50 microseconds, 5 times), 783 (25%) hatched out. Four percent of the hatchlings were transgenic.