Improving Flavonoid Accumulation of Bioreactor-Cultured Adventitious Roots in Oplopanax elatus Using Yeast Extract.

Oplopanax elatus is an endangered medicinal plant, and adventitious root (AR) culture is an effective way to obtain its raw materials. Yeast extract (YE) is a lower-price elicitor and can efficiently promote metabolite synthesis. In this study, the bioreactor-cultured O. elatus ARs were treated with YE in a suspension culture system to investigate the elicitation effect of YE on flavonoid accumulation, serving for further industrial production. Among YE concentrations (25-250 mg/L), 100 mg/L YE was the most suitable for increasing the flavonoid accumulation. The ARs with various ages (35-, 40-, and 45-day-old) responded differently to YE stimulation, where the highest flavonoid accumulation was found when 35-day-old ARs were treated with 100 mg/L YE. After YE treatment, the flavonoid content increased, peaked at 4 days, and then decreased. By comparison, the flavonoid content and antioxidant activities in the YE group were obviously higher than those in the control. Subsequently, the flavonoids of ARs were extracted by flash extraction, where the optimized extraction process was: 63% ethanol, 69 s of extraction time, and a 57 mL/g liquid-material ratio. The findings provide a reference for the further industrial production of flavonoid-enriched O. elatus ARs, and the cultured ARs have potential application for the future production of products.

Adventitious root cultures of Oplopanax elatus inhibit LPS-induced inflammation via suppressing MAPK and NF-κB signaling pathways.

Bioreactor-cultured adventitious roots (ARs) of the endangered medicinal plant Oplopanax elatus Nakai is a novel alternative plant material. To utilize ARs in the product production, the present study investigated the anti-inflammatory effect of O. elatus ARs. In the in vivo experiment, lipopolysaccharide (LPS)-induced acute lung injury disease model was established and several inflammatory indexes were determined. For the LPS-stimulated mice, after pretreatment of AR crude extract (200 mg/kg), cell infiltration in lungs was decreased, the production of proinflammatory mediators, including nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin (IL)-6, and 1ß in the bronchoalveolar lavage fluid was evidently reduced, which indicated that O. elatus ARs had an anti-inflammatory effect. In the in vitro experiment, ethyl acetate (EtOAc) fractions (12.5, 25, and 50 µg/mL) were used to treat LPS-induced peritoneal macrophages (PMs) of mice. The production of NO, prostaglandin E2, TNF-α, IL-6, and IL-1ß in LPS-stimulated PMs was obviously inhibited (p < 0.05) after pretreatment with EtOAc fractions, and the expression of the inducible nitric oxide synthase and cyclooxygenase were also suppressed. To clarify the anti-inflammatory mechanism, effects of EtOAc fraction on changes of proteins related to the pathways of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB) were investigated. The phosphorylation of extracellular regulated protein kinases, c-jun n-terminal kinase, and p38 MAPK in LPS-induced PMs was inhibited after pretreatment of EtOAc fractions. In addition, EtOAc fractions enhanced inhibitor of nuclear factor-kappa B-α expression and decreased nuclear translocation of p65 NF-κB. Thus, EtOAc from O. elatus ARs is involved in regulating MAKP and NF-κB signaling pathways to inhibit LPS-induced inflammation.

Production of Rare Ginsenosides Rg3 and Rh2 by Endophytic Bacteria from Panax ginseng.

The ginsenosides Rh2 and Rg3 induce tumor cell apoptosis, inhibit tumor cell proliferation, and restrain tumor invasion and metastasis. Despite Rh2 and Rg3 having versatile pharmacological activities, contents of them in natural ginseng are extremely low. To produce ginsenosides Rh2 and Rg3, the saponin-producing capacity of endophytic bacteria isolated from Panax ginseng was investigated. In this work, 81 endophytic bacteria isolates were taken from ginseng roots by tissue separation methods. Among them, strain PDA-2 showed the highest capacity to produce the rare ginsenosides; the concentrations of rare ginsenosides Rg3 and Rh2 reached 62.20 and 18.60 mg/L, respectively. On the basis of phylogenetic analysis, it was found that strain PDA-2 belongs to the genus Agrobacterium and was very close to Agrobacterium rhizogenes.

Recent progress and perspectives in biotrickling filters for VOCs and odorous gases treatment.

Pollution caused by volatile organic compounds (VOCs) and odorous pollutants in the air can produce severe environmental problems. In recent years, the emission control of VOCs and odorous pollutants has become a crucial issue owing to the adverse effect on humans and the environment. For treating these compounds, biotrickling filter (BTF) technology acts as an environment friendly and cost-effective alternative to conventional air pollution control technologies. Besides, low concentration of VOCs and odorous pollutants can also be effectively removed using BTF systems. However, the VOCs and odorants removal performance by BTF may be limited by the hydrophobicity, toxicity, and low bioavailability of these pollutants. To solve these problems, this review summarizes the design, mechanism, and common analytical methods of recent BTF advances. In addition, the operating conditions, mass transfer, packing materials and microorganisms (which are the critical parameters in a BTF system) were evaluated and discussed in view of improving the removal performance of BTFs. Further research on these specific topics, together with the combination of BTF technology with other technologies, should improve the removal performance of BTFs.

Endophytic Bacteria Isolated from Panax ginseng Improves Ginsenoside Accumulation in Adventitious Ginseng Root Culture.

Ginsenoside is the most important secondary metabolite of ginseng. Natural sources of wild ginseng have been overexploited. Although root culture could reduce the length of the growth cycle of ginseng, the number of ginsenosides is fewer and their contents are lower in adventitious roots of ginseng than that in ginseng cultivated in the field. In this study, we investigated the effects of endophytic bacterial elicitors on biomass and ginsenoside production in adventitious roots cultures of Panax ginseng. Endophyte LB 5-3 as an elicitor could increase biomass and ginsenoside accumulation in ginseng adventitious root culture. After 6 days elicitation with a 10.0 mL of strain LB 5-3, the content of total ginsenoside was 2.026 mg g-1 which was four times more than that in unchallenged roots. The combination of methyl jasmonate and strain LB 5-3 had a negative effect on ginseng adventitious root growth and ginsenoside production. The genomic DNA of strain LB 5-3 was sequenced, and was found to be most closely related to Bacillus altitudinis (KX230132.1). The challenged ginseng adventitious root extracts exerted inhibitory effect against the HepG2 cells, which IC50 value was 0.94 mg mL-1.

Microbial transformation of ginsenosides extracted from Panax ginseng adventitious roots in an airlift bioreactor

Background: Ginsenoside is the most important secondary metabolite in ginseng. Natural sources of wild ginseng have been overexploited. Although root culture can reduce the length of the growth cycle of ginseng, the number of species of ginsenosides is reduced and their contents are lower in the adventitious roots of ginseng than in the roots of ginseng cultivated in the field. Results: In this study, 147 strains of ß-glucosidase-producing microorganisms were isolated from soil. Of these, strain K35 showed excellent activity for converting major ginsenosides into rare ginsenosides, and a NCBI BLAST of its 16S rDNA gene sequence showed that it was most closely related to Penicillium sp. (HQ608083.1). Strain K35 was used to ferment the adventitious root extract, and the fermentation products were analyzed by high-performance liquid chromatography. The results showed that the content of the rare ginsenoside CK was 0.253 mg mL-1 under the optimal converting conditions of 9 d of fermentation at pH 7.0 in LL medium, which was significantly higher than that in the adventitious roots of ginseng. Conclusion: These findings may not only solve the problem of low productivity of metabolite in ginseng root culture but may also result in the development of a new valuable method of manufacturing ginsenoside CK.

Fermentation of ginseng extracts by Penicillium simplicissimum GS33 and anti-ovarian cancer activity of fermented products.

A total of 58 isolates of ß-glucosidase-producing microorganisms were isolated from soil around the wild ginseng roots under forest using Esculin-R2A agar. Among these isolates, strain GS33 showed a strong ability to convert ginsenosides Rb1, Rb2, Rc, and Rd into F2, Rg3, C-K, and convert ginsenoside Rg1 into Rh1, and F1. Fermented ginseng products can inhibit ES-2 cells growth and the IC50 value was 0.73 mg ml⁻¹. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain GS33 belongs to the genus Penicillium and is most closely related to Penicillium simplicissimum (99 %).

Co-transformation of Panax major ginsenosides Rb1 and Rg1 to minor ginsenosides C-K and F1 by Cladosporium cladosporioides.

Rb1 and Rg1 are the major ginsenosides in protopanaxadiol and protopanaxatriol. Their content in ginsenosides was 23.8 and 17.6%, respectively. A total of 22 isolates of ß-glucosidase producing microorganisms were isolated from the soil of a ginseng field using Esculin-R2A agar. Among these isolates, the strain GH21 showed the strongest activities to convert ginsenoside Rb1 and Rg1 to minor ginsenosides compound-K and F1, respectively. Ginsenosides Rb1 and Rg1 bioconversion rates were 74.2 and 89.3%, respectively. Meanwhile, the results demonstrated that the ginsenoside Rg1 could change the biotransformation pathway of ginsenoside Rb1 by inhibiting the formation of the intermediate metabolite gypenoside-XVII. GH21 was identified as a Cladosporium cladosporioides species based on the internal transcribed spacers (ITS) ITS1-5.8S-ITS2 rRNA gene sequences constructed phylogenetic trees.

Tumebacillus ginsengisoli sp. nov., isolated from soil of a ginseng field.

A gram-reaction-positive, rod-shaped, spore-forming bacterium, designated Gsoil 1105(T), was isolated from soil of a ginseng field in Pocheon Province in South Korea and characterized in order to determine its taxonomic position. Comparative analysis of the 16S rRNA gene sequence showed that the isolate belongs to the order Bacillales, showing the highest level of sequence similarity with respect to Tumebacillus permanentifrigoris Eur1 9.5(T) (94.6 %). The phylogenetic distances from other described species with validly published names within the order Bacillales were greater than 9.0 %. Strain Gsoil 1105(T) had a genomic DNA G+C content of 55.6 mol% and menaquinone 7 (MK-7) as the major respiratory quinone. The major fatty acids were iso-C(15 : 0) and anteiso-C(15 : 0). On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil 1105(T) represents a novel species of the genus Tumebacillus, for which the name Tumebacillus ginsengisoli sp. nov. is proposed. The type strain is Gsoil 1105(T) ( = KCTC 13942(T)  = DSM 18389(T)).

Aeromicrobium panaciterrae sp. nov., isolated from soil of a ginseng field in South Korea.

A Gram-positive, rod-shaped, non-spore-forming and strictly aerobic bacterium (Gsoil 161(T)) was isolated from soil of a ginseng field in Pocheon Province, South Korea. The novel isolate was characterized using a polyphasic approach in order to determine its taxonomic position. On the basis of 16S rRNA gene sequence similarity, strain Gsoil 161(T) was shown to belong to the family Nocardioidaceae and was related to Aeromicrobium marinum (98.0 % similarity to the type strain), Aeromicrobium alkaliterrae (97.6 %), Aeromicrobium fastidiosum (97.0 %) and Aeromicrobium erythreum (96.7 %); the sequence similarity with other species within the family was less than 94.4 %. It was characterized chemotaxonomically as having ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan, MK-9(H(4)) as the predominant menaquinone and C(16 : 0), 10-methyl C(18 : 0) (tuberculostearic acid), C(16 : 0) 2-OH, 10-methyl C(17 : 0) and 10-methyl-C(16 : 0) as the major fatty acids. The G+C content of the genomic DNA was 65.5 mol%. These chemotaxonomic properties and phenotypic characteristics support the affiliation of strain Gsoil 161(T) to the genus Aeromicrobium. Results of physiological and biochemical tests enabled strain Gsoil 161(T) to be differentiated genotypically and phenotypically from currently known Aeromicrobium species. Therefore, strain Gsoil 161(T) represents a novel species, for which the name Aeromicrobium panaciterrae sp. nov. is proposed. The type strain is strain Gsoil 161(T) (=KCTC 19131(T)=DSM 17939(T)=CCUG 52476(T)).

Microlunatus ginsengisoli sp. nov., isolated from soil of a ginseng field.

A Gram-positive, aerobic, coccus-shaped, non-endospore-forming bacterium (Gsoil 633(T)) was isolated from soil from a ginseng field in Pocheon province in South Korea. The novel isolate was characterized in order to determine its taxonomic position. On the basis of 16S rRNA gene sequence similarities, strain Gsoil 633(T) was shown to belong to the family Propionibacteriaceae. The closest phylogenetic relative was Microlunatus phosphovorus DSM 19555(T), with 96.1 % sequence similarity; the sequence similarity to other members of the family was less than 95.4 %. The isolate was characterized chemotaxonomically as having ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan, MK-9(H(4)) as the predominant menaquinone and anteiso-C(15 : 0), iso-C(15 : 0) and iso-C(16 : 0) as the major fatty acids. The G+C content of the genomic DNA was 69.8 mol%. The morphological and chemotaxonomic properties of the isolate were consistent with those of M. phosphovorus, but the results of physiological and biochemical tests allowed the phenotypic differentiation of strain Gsoil 633(T) from this species. Therefore, strain Gsoil 633(T) represents a novel species, for which the name Microlunatus ginsengisoli sp. nov. is proposed. The type strain is Gsoil 633(T) (=KCTC 13940(T)=DSM 17942(T)).

Odorous swine wastewater treatment by purple non-sulfur bacteria, Rhodopseudomonas palustris, isolated from eutrophicated ponds.

Three strains of phototrophic, purple, non-sulfur bacteria, isolated from eutrophic ponds, were used to treat odorous swine wastewater. One isolate, Rhodopseudomonas palustris, when cultured in swine wastewater without supplementation for 7 d, removed odorous organic acids (170 mg l(-1)), COD (10,000 mg l(-1)) and phosphate (180 mg l(-1)).