First record of Ginkgo-toothed beaked whale (Mesoplodon ginkgodens) stranded in Korea.

Two stranded whales were found dead on the coast of Jeju, South Korea. Based on the outer appearance and autopsy findings, one was determined to be an adult and the other a calf. The carcasses were dissected for species identification and pathological examination. A genetic analysis was performed, and the morphological characteristics of the skull observed. Then, 448 bp of the 5' half of the mitochondrial (mt) DNA control region and 413 bp of the mtDNA cytochrome b gene were sequenced. A BLAST search revealed that the whales were ginkgo-toothed beaked whales (Mesoplodon ginkgodens). Morphological comparison of the adult skull with the holotype specimen confirmed the result. This is the first record of a stranded ginkgo-toothed beaked whale in Korea.

ß-glucosidase from Penicillium aculeatum hydrolyzes exo-, 3-O-, and 6-O-ß-glucosides but not 20-O-ß-glucoside and other glycosides of ginsenosides.

A novel ß-glucosidase from Penicillium aculeatum was purified as a single 110.5-kDa band on SDS-PAGE with a specific activity of 75.4 U mg⁻¹ by salt precipitation and Hi-Trap Q HP and Resource Q ion exchange chromatographies. The purified enzyme was identified as a member of the glycoside hydrolase 3 family based on its amino acid sequence. The hydrolysis activity for p-nitrophenyl-ß-D-glucopyranoside was optimal at pH 4.5 and 70 °C with a half-life of 55 h. The enzyme hydrolyzed exo-, 3-O-, and 6-O-ß-glucosides but not 20-O-ß-glucoside and other glycosides of ginsenosides. Because of the novel specificity, this enzyme had the transformation pathways for ginsenosides: Rb1 → Rd → F2 → compound K, Rb2 → compound O → compound Y, Rc → compound Mc1 → compound Mc, Rg3 → Rh2 → aglycone protopanaxadiol (APPD), Rg1 → F1, and Rf → Rh1 → aglycone protopanaxatriol (APPT). Under the optimum conditions, the enzyme converted 0.5 mM Rb2, Rc, Rd, Rg3, Rg1, and Rf to 0.49 mM compound Y, 0.49 mM compound Mc, 0.47 mM compound K, 0.23 mM APPD, 0.49 mM F1, and 0.44 mM APPT after 6 h, respectively.

Ginsenoside F1 production from ginsenoside Rg1 by a purified ß-glucosidase from Fusarium moniliforme var. subglutinans.

Fusarium moniliforme var. subglutinans was selected from among 100 strains of fungi for producing ginsenoside F(1) from ginsenoside Rg(1). The enzyme responsible was purified as a single 85 kDa band with a specific activity of 136 U mg(-1). It hydrolysed glucose-linked ginsenosides Rb(1), Rd and Rg(1) but not for other monosaccharide-linked ginsenosides, Rb(2), Rc, R(1), and Re. Under the optimum conditions of pH 6.0, 50 °C, 30 U l(-1) of enzyme, and 5 mg Rg(1) ml(-1), 4 mg F(1) ml(-1) was produced after 4 h, with a molar yield of 100% and a productivity of 1 g l(-1) h(-1). This represents the highest productivity and conversion yield of F(1) yet reported.

Production of aglycon protopanaxadiol via compound K by a thermostable ß-glycosidase from Pyrococcus furiosus.

The production of compound K and aglycon protopanaxadiol (APPD) from ginsenoside Rd and ginseng root extract was performed using a recombinant ß-glycosidase from Pyrococcus furiosus. The activity for Rd was optimal at pH 5.5 and 95°C with a half-life of 68 h at 95°C. ß-Glycosidase converted Rb(1), Rb(2), Rc, and Rd to APPD via compound K. With increases in the enzyme activity, the productivities of compound K and APPD increased. The substrate concentration was optimal at 4.0 mM Rd or 10% (w/v) ginseng root extract; 4 mM of Rd was converted to 3.3 mM compound K with a yield of 82.5% (mol/mol) and a productivity of 2,010 mg l(-1) h(-1) at 1 h and was hydrolyzed completely to APPD with 364 mg l(-1) h(-1) after 5 h. Rb(1), Rb(2), Rc, and Rd at 3.9 mM in 10% ginseng root extract were converted to 3.1 mM compound K with 79.5% and 1,610 mg l(-1) h(-1) at 1.2 h and were hydrolyzed completely to APPD with 300 mg l(-1) h(-1) after 6 h. The concentrations and productivities of compound K and APPD in the present study are the highest ever reported.

Immobilization stress induces endothelial dysfunction by oxidative stress via the activation of the angiotensin II/its type I receptor pathway.

OBJECTIVE: Psychological stress has been shown to contribute to the development of atherosclerosis; however its underlying mechanism has not been clearly elucidated. We here studied the mechanism by which immobilization stress causes endothelial dysfunction with specific aim of identifying the role of angiotensin II and its type I (AT(1)) receptor signaling pathway. METHODS AND RESULTS: Rats (n=30) were subjected to immobilization stress (120 min/day) for 14 days using a restrainer. During immobilized period, rats were orally administrated with or without the angiotensin converting enzyme (ACE) inhibitor ramipril (3 mg/kg/day, n=10) or AT(1) receptor inhibitor losartan (9 mg/kg/day, n=10). Immobilization significantly increased systolic blood pressure and decreased acetylcholine-induced ex vivo relaxation of arteries compared with those of control animals (n=10). Immobilization increased the plasma levels of angiotensin II and ACE activity that were inhibited by treatment with ramipril, but not losartan. Furthermore, immobilization increased the plasma level of malondialdehyde and expression of gp91(phox) and Rho-associated kinase-1 in arteries, and decreased the arterial eNOS mRNA and oxidized products of NO (nitrite plus nitrate). These functional and biochemical alterations induced by immobilization were significantly reversed by administration of ramipril or losartan. CONCLUSIONS: Immobilization stress induces vascular oxidative stress by activating the angiotensin II/AT(1) receptor signaling pathway, thereby provoking endothelial dysfunction which can contribute to the development of atherosclerosis and hypertension.

Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases.

Ginsenosides are the principal components responsible for the pharmaceutical activities of ginseng. The minor ginsenosides, which are also pharmaceutically active, can be produced via the hydrolysis of the sugar moieties in the major ginsenosides using acid hydrolytic, heating, microbial, and enzymatic transformation techniques. The enzymatic method has a profound potential for ginsenoside transformation, owing to its high specificity, yield, and productivity, and this method is increasingly being recognized as a useful tool in structural modification and metabolism studies. In this article, the transformation methods of ginsenosides, the characterization of microbial glycosidases with ginsenoside hydrolyzing activities, and the enzymatic production of minor ginsenosides are reviewed. Moreover, the conversions of ginsenosides using cell extracts from food microorganisms and recombinant thermostable beta-D-glycosidases are proposed as feasible methods for use in industrial processes.

Discrimination of animal species using polymorphisms of the nuclear gene zinc finger protein 238.

We screened 3750 single exonic genes listed in the intronless genes in the eukaryotes (SEGE) database and performed bioinformatic analyses to identify candidate genes for new species-specific markers. A set of PCR primers for the conserved regions of ZNF238 was developed and used to amplify the 823 bp DNA fragment. We compared nucleotide variations of the PCR products among 20 species plus two subspecies of animals, which led to the identification of interspecies nucleotide variations. To establish a simple method for the analysis of species-specific DNA polymorphisms using ZNF238, we developed a PCR-RFLP method using HhaI and HpyCH4IV restriction enzymes for 13 species. For the remaining species, the direct sequencing of PCR products provided additional SNPs, enabling precise species classification. As a result, we report here that a new nuclear DNA marker, ZNF238, can be used to increase the accuracy of species identification among euteleostomi (bony vertebrates).

Identification of single-nucleotide polymorphisms of the prion protein gene in sika deer (Cervus nippon laiouanus).

Polymorphisms of the prion protein gene (PRNP) have been detected in several cervid species. In order to confirm the genetic variations, this study examined the DNA sequences of the PRNP obtained from 33 captive sika deer (Cervus nippon laiouanus) in Korea. A total of three single-nucleotide polymorphisms (SNPs) at codons 100, 136 and 226 in the PRNP of the sika deer were identified. The polymorphic site located at codon 100 has not been reported. The SNPs detected at codons 100 and 226 induced amino acid substitutions. The SNP at codon 136 was a silent mutation that does not induce any amino acid change. The genotype and allele frequencies were determined for each of the SNPs.