Terrabacter ginsengisoli sp. nov., isolated from ginseng cultivating soil.

A Gram-positive, strictly aerobic, nonmotile, yellowish, coccus-rod-shaped bacterium (designated Gsoil 653T) isolated from ginseng cultivating soil was characterized using a polyphasic approach to clarify its taxonomic position. The strain Gsoil 653T exhibited optimal growth at pH 7.0 on R2A agar medium at 30°C. Phylogenetic analysis based on 16S rRNA gene sequence similarities, indicated that Gsoil 653T belongs to the genus Terrabacter of the family Humibacillus, and was closely related to Terrabacter tumescens DSM 20308T (98.9%), Terrabacter carboxydivorans PY2T (98.9%), Terrabacter terrigena ON10T (98.8%), Terrabacter terrae PPLBT (98.6%), and Terrabacter lapilli LR-26T (98.6%). The DNA G + C content was 70.5 mol%. The major quinone was MK-8(H4). The primary polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidyl-ethanolamine. The predominant fatty acids were iso-C15:0, iso-C16:0, iso-C14:0, and anteiso-C15:0, as in the case of genus Terrabacter, thereby supporting the categorization of strain Gsoil 653T. However, the DNA-DNA relatedness between Gsoil 653T and closely related strains of Terrabacter species was low at less than 31%. Moreover, strain Gsoil 653T could be both genotypically and phenotypically distinguished from the recognized species of the genus Terrabacter. This isolate, therefore, represents a novel species, for which the name Terrabacter ginsengisoli sp. nov. is proposed with the type strain Gsoil 653T (= KACC 19444T = LMG 30325T).

Baekduia soli gen. nov., sp. nov., a novel bacterium isolated from the soil of Baekdu Mountain and proposal of a novel family name, Baekduiaceae fam. nov.

A taxonomic study was conducted on BR7-21T, a bacterial strain isolated from the soil of a ginseng field in Baekdu Mountain. Comparative studies of the 16S rRNA gene sequence showed that the isolate was most closely related to Conexibacter woesei DSM 14684T, Solirubrobacter pauli ATCC BAA-492T, Patulibacter minatonensis JCM 12834T, with 93.8%, 92.4%, and 91.5% sequence similarity, respectively; each genus represented a family in the order Solirubrobacterales. Strain BR7-21T was Gram-reaction positive, non-spore forming, aerobic, non-motile, and short rod-shaped. It grew well on half-strength R2A medium. The G + C content of the genomic DNA was 73.9%. It contained meso-diaminopimelic acid in the cell wall and the major menaquinones were MK-7(H4) and MK-8(H4). The major fatty acids were summarized as (C16:1ω7c/iso-C15:0 2-OH), iso-C16:0, and C17:0 cyclo. On the basis of polyphasic evidence, it was proposed that strain BR7-21T should be placed in a new genus and species, for which the name Baekduia soli gen. nov., sp. nov. was proposed with the type strain BR7-21T (= KCTC 22257T = LMG 24797T). The family Baekduiaceae fam. nov. is proposed to encompass the genus Baekduia gen. nov.

Mucilaginibacter pocheonensis sp. nov., with ginsenoside-converting activity, isolated from soil of a ginseng-cultivating field.

A Gram-reaction-negative, aerobic, heterotrophic, non-motile, non-spore-forming, rod-shaped bacterial strain, designated Gsoil 032T, was isolated from soil of a ginseng field in Pocheon Province, South Korea, and its taxonomic position was investigated by using a polyphasic approach. Strain Gsoil 032T grew at 10-42 °C and at pH 5.0-10.0 on R2A agar medium. Strain Gsoil 032T possessed ß-glucosidase activity, which was responsible for its ability to transform ginsenoside Rb1 (one of the dominant active components of ginseng) to compound K. On the basis of 16S rRNA gene sequence similarity, strain Gsoil 032T was shown to belong to the family Sphingobacteriaceae and to be related to Mucilaginibacter sabulilitoris SMS-12T (97.6 % sequence similarity) and Mucilaginibacter lappiensis ANJLI2T (97.1 %) The G+C content of the genomic DNA was 44.4 mol%. The predominant respiratory quinone was menaquinone MK-7 and the major fatty acids were summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c), iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipid detected was phosphatidylethanolamine, while the minor polar lipids were various unidentified aminophospholipids, unidentified phospholipids and unidentified polar lipids. DNA and chemotaxonomic data supported the affiliation of strain Gsoil 032T to the genus Mucilaginibacter. Strain Gsoil 032T could be differentiated genotypically and phenotypically from recognized species of the genus Mucilaginibacter. The isolate therefore represents a novel species, for which the name Mucilaginibacter pocheonensis sp. nov. is proposed, with the type strain Gsoil 032T (=KCTC 12641T=LMG 23495T).

Flavisolibacter ginsenosidimutans sp. nov., with ginsenoside-converting activity isolated from soil used for cultivating ginseng.

A Gram-reaction-negative, aerobic, non-motile and rod-shaped bacterial strain designated Gsoil 636T was isolated from soil of a ginseng cultivation field in Pocheon Province, South Korea and its taxonomic position was investigated using a polyphasic approach. Gsoil 636T grew at 18-30 °C and at pH 6.0-8.0 on R2A medium. Gsoil 636T possessed ß-glucosidase activity, which was responsible for its ability to transform ginsenoside Rb1 (ones of the dominant active components of ginseng) to F2. On the basis of 16S rRNA gene sequence similarity, Gsoil 636T was shown to belong to the family Chitinophagaceae and to be related to Flavisolibacter ginsengiterrae Gsoil 492T (96.7 % sequence similarity), Flavisolibacter ginsengisoli Gsoil 643T (96.6 %) and Flavisolibacter rigui 02SUJ3T (96.6 %). The G+C content of the genomic DNA was 48.9 %. The predominant respiratory quinone was MK-7 and the major fatty acids were iso-C15 : 0, summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c) and iso-C17 : 0 3-OH. DNA and chemotaxonomic data supported the affiliation of Gsoil 636T to the genus Flavisolibacter. Gsoil 636T could be differentiated genotypically and phenotypically from the species of the genus Flavisolibacter with validly published names. The isolate therefore represents a novel species, for which the name Flavisolibacter ginsenosidimutans sp. nov. is proposed, with the type strain Gsoil 636T (KCTC 22818T = JCM 18197T = KACC 14277T).

20(S)-Ginsenoside Rh2 as aldose reductase inhibitor from Panax ginseng.

The root of Panax ginseng C. A. Meyer (Araliaceae) is a well-known herbal medicine in East Asia. The major bioactive metabolites in this root are commonly identified as ginsenosides. A series of ginsenosides were determined for in vitro human recombinant aldose reductase. This Letter aims to clarify the structural requirement for aldose reductase inhibition. We discovered that only ginsenoside 20(S)-Rh2 showed potent against aldose reductase, with an IC50 of 147.3 µM. These results implied that the stereochemistry of the hydroxyl group at C-20 may play an important role in aldose reductase inhibition. An understanding of these requirements is considered necessary in order to develop a new type of aldose reductase inhibitor. Furthermore, P. ginseng might be an important herbal medicine in preventing diabetic complications.

Identification and characterization of a novel Terrabacter ginsenosidimutans sp. nov. beta-glucosidase that transforms ginsenoside Rb1 into the rare gypenosides XVII and LXXV.

A new beta-glucosidase from a novel strain of Terrabacter ginsenosidimutans (Gsoil 3082(T)) obtained from the soil of a ginseng farm was characterized, and the gene, bgpA (1,947 bp), was cloned in Escherichia coli. The enzyme catalyzed the conversion of ginsenoside Rb1 {3-O-[beta-D-glucopyranosyl-(1-2)-beta-D-glucopyranosyl]-20-O-[beta-D-glucopyranosyl-(1-6)-beta-D-glucopyranosyl]-20(S)-protopanaxadiol} to the more pharmacologically active rare ginsenosides gypenoside XVII {3-O-beta-D-glucopyranosyl-20-O-[beta-D-glucopyranosyl-(1-6)-beta-D-glucopyranosyl]-20(S)-protopanaxadiol}, gypenoside LXXV {20-O-[beta-v-glucopyranosyl-(1-6)-beta-D-glucopyranosyl]-20(S)-protopanaxadiol}, and C-K [20-O-(beta-D-glucopyranosyl)-20(S)-protopanaxadiol]. A BLAST search of the bgpA sequence revealed significant homology to family 3 glycoside hydrolases. Expressed in E. coli, beta-glucosidase had apparent K(m) values of 4.2 +/- 0.8 and 0.14 +/- 0.05 mM and V(max) values of 100.6 +/- 17.1 and 329 +/- 31 micromol x min(-1) x mg of protein(-1) against p-nitrophenyl-beta-D-glucopyranoside and Rb1, respectively. The enzyme catalyzed the hydrolysis of the two glucose moieties attached to the C-3 position of ginsenoside Rb1, and the outer glucose attached to the C-20 position at pH 7.0 and 37 degrees C. These cleavages occurred in a defined order, with the outer glucose of C-3 cleaved first, followed by the inner glucose of C-3, and finally the outer glucose of C-20. These results indicated that BgpA selectively and sequentially converts ginsenoside Rb1 to the rare ginsenosides gypenoside XVII, gypenoside LXXV, and then C-K. Herein is the first report of the cloning and characterization of a novel ginsenoside-transforming beta-glucosidase of the glycoside hydrolase family 3.

Stereospecificity of hydroxyl group at C-20 in antiproliferative action of ginsenoside Rh2 on prostate cancer cells.

Prostate cancer is the fifth most common neoplasm worldwide, and the second most common cancer among men. Ginsenosides, the main component of ginseng, have been known for their medicinal effects such as anti-inflammatory and anti-proliferative activities. In this study, we investigated the inhibitory effects of ginsenosides (ginsenoside 20(R)-Rh2 and ginsenoside 20(S)-Rh2) on prostate cancer cells in vitro. Only ginsenoside 20(S)-Rh2 showed proliferation inhibition on androgen-dependent and -independent prostate cancer cells. These results implied that the stereochemistry of the hydroxyl group at C-20 may play an important role in antitumor activities.

[Microorganisms of special herb-glycosidases and their fermentation, enzyme properties].

Herb-glycosides are main active elements of Zhongcaoyao (Chinese traditional medicines, Chinese medical herbs). However, the herb-glycoside structures are not optimal active structure for the human bodies. After orally taken up, the herb-glycosides of Zhongcaoyao could be changed into other more active structures by the digestive system such as enzymes and intestinal microorganisms; then degraded and absorbed in the human body and play the real role of pharmic effect; but only a small amount could be changed and controlled by circadian state of the human body. If this biochange of herb-glycosides to more active structures in vivo was finished in vitro, it is very useful for the development of the Chinese traditional medicines, new plant medicines, health food, and function cosmetics. To biotransformate herb-glycosides to more active structure, this paper introduced the studies of author's team on the new microorganism isolation of the special herb-glycosidases and enzyme fermentation, the special enzyme purification and characterization.

Emticicia ginsengisoli sp. nov., a species of the family 'Flexibacteraceae' isolated from soil of a ginseng field.

A taxonomic study was carried out on Gsoil 085T, a bacterial strain isolated from a soil sample from a ginseng field in Pocheon Province (South Korea). Comparative 16S rRNA gene sequence studies showed a clear affiliation of this bacterium to the family 'Flexibacteraceae' and indicated that the closest relative was Emticicia oligotrophica GPTSA100-15T (94.6 % sequence similarity). Lower sequence similarities (<83.0 %) were found with respect to all other recognized species of the family 'Flexibacteraceae'. Strain Gsoil 085T was found to be Gram-negative, strictly aerobic, non-motile and rod-shaped. It grew well on half-strength R2A medium but did not utilize a broad range of carbon sources. The G+C content of the genomic DNA was 40.5 mol%. The major fatty acids were summed feature 4 (iso-C15 : 0 2-OH and/or C16 : 1omega7c, 45.4 %), iso-C15 : 0 (13.9 %), C16 : 1omega5c (8.6 %) and C16 : 0 (8.2 %). On the basis of evidence from the polyphasic study, strain Gsoil 085T represents a novel species of the genus Emticicia, for which the name Emticicia ginsengisoli sp. nov. is proposed. The type strain is Gsoil 085T (=KCTC 12588T =LMG 23396T).

[Establishment of high performance liquid chromatographic fingerprint of Fructus schisandrae chiensis].

The chromatographic fingerprint was established for evaluating and controlling the quality of Fructus schisandrae chinensis by reversed-phase high performance liquid chromatography (RP-HPLC). Ten different original samples were analyzed, and the chromatographic fingerprint of Fructus schisandrae chinensis was constructed with 26 fingerprint peaks, among which 19 peaks were common fingerprint peaks and 5 main compounds were identified using the Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine (Version 2004 A) recommended by State Food & Drug Administration. Methods of vector cosine and correlation coefficient were used to calculate the similarity of ten samples, and satisfactory results were obtained. The chromatographic fingerprint can be used to differentiate the raw materials from different sources and be served as a powerful tool for the further quality control of Fructus schisandrae chinensis.

Purification and characterization of new special ginsenosidase hydrolyzing multi-glycisides of protopanaxadiol ginsenosides, ginsenosidase type I.

In this paper, the new type ginsenosidase which hydrolyzing multi-glycosides of ginsenoside, named ginsenoside type I from Aspergillus sp.g48p strain was isolated, characterized and generally described. The enzyme molecular weight was about 80 kDa. Ginsenosidase type I can hydrolyze different glycoside of protopanaxadiol type ginsenosides (PPD); i.e., can hydrolyze the 3(carbon)-O-beta-glucoside of Rb1, Rb2, Rb3, Rc, Rd; can hydrolyze 20(carbon)-O-beta-glucoside of Rb1, 20-O-beta-xyloside of Rb3, 20-O-alpha-arabinoside(p) of Rb2 and 20-O-alpha-arabinoside(f) of Rc to produce mainly F2, compound-K (C-K) and small Rh2, but can not hydrolyze the glycosides of protopanaxatriol type ginsenoside (PPT) such as Re, Rf, Rg1. So, when the ginsenosidase type I hydrolyzed ginsenosides, the enzyme selected ginsenoside-aglycone type, can hydrolyze different glycosides of PPD type ginsenoside; however no selected glycoside type, can hydrolyze multi-glycosides of PPD type ginsenosides. These properties were novel properties, and differentiated with the other previously described glycosidases.