Streptomyces zingiberis sp. nov., an endophytic actinobacterium isolated from the root tissue of Zingiber montanum.

An endophytic actinobacterium, designated strain PLAI 1-29T, was isolated from the root tissue of Zingiber montanum collected from Pathum Thani province, Thailand. Strain PLAI 1-29T was characterized using a polyphasic taxonomic approach. It typically exhibited morphological and chemotaxonomic properties of the genus Streptomyces. Strain PLAI 1-29T produced a spiral spore chain on aerial mycelium and grew at 15-40 °C, pH 6-10 on International Streptomyces Project 2 agar. The maximum NaCl concentration for growth was 9 % (w/v). Cells of strain PLAI 1-29T presented ll-diaminopimelic acid, arabinose, galactose and ribose. The detected phospholipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. The major menaquinones were MK-9(H6) and MK-9(H8). The major cellular fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The genome-based taxonomic details revealed the assignment of strain PLAI 1-29T to the genus Streptomyces and exhibited low threshold values for the delineation of a novel species by average nucleotide identity-blast (84.0%), average amino acid identity (80.0%) and digital DNA-DNA hybridization (27.6%) with its closest type strain, Streptomyces xinghaiensis S187T. Furthermore, several differential physiological and biochemical characteristics were detected between strain PLAI 1-29T and the closest type strain. Based on the combined phenotypic and genomic features, strain PLAI 1-29T (=TBRC 7645T=NBRC 113170T) is considered to represent a new Streptomyces species, for which we propose the name Streptomyces zingiberis sp. nov.

Mass spectral analysis of secondary metabolites from Zingiber montanum rhizome extract using UHPLC-HR-ESI-QTOF-MS/MS.

INTRODUCTION: Zingiber montanum (J.Koenig) Link ex A.Dietr. is a popular medicinal plant in Thailand. Its rhizomes have been used as an ingredient in various Thai traditional medicine formulas. While many reports have focused on the chemical constituents and biological activities of this plant, a comprehensive study on secondary metabolite profiling using tandem mass spectrometry has, to this point, never been documented. OBJECTIVE: To analyze the chemical constituents in Z. montanum rhizomes using ultra-high performance liquid chromatography coupled with ultra-high-resolution electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-HR-ESI-QTOF-MS/MS) analyses and to utilize the characteristic fragmentation patterns of these compounds to facilitate their identification. METHODOLOGY: UHPLC-HR-ESI-QTOF-MS/MS in positive ion mode was used for chemical identification of secondary metabolites from the ethanolic extract of the plant material. MS/MS data of some known reference compounds, together with detailed fragmentation pattern information of several compounds obtained from the crude extract, were used to elucidate their chemical structures. RESULTS: In this work, one benzaldehyde, ten phenylbutenoid monomers, six curcuminoids, and nine phenylbutenoid dimers were assigned based on their characteristic fragment ions. Among these compounds, 2-(3,4-dimethoxystyryl)oxirane was tentatively suggested as a potential new compound. Several characteristic fragment ions from these compounds were assigned and the relative ion abundance of these was also used to differentiate the chemical structures of compounds having the same molecular mass. CONCLUSIONS: The results will benefit future high-throughput screening of bioactive compounds and method development for the quality control of raw materials and herbal drugs derived from Z. montanum rhizome extracts.

Secondary metabolites content and essential oil composition of in vitro cultures of Zingiber montanum (Koenig) Link ex A. Dietr.

OBJECTIVES: To determine the secondary metabolite content, antioxidant and phenylalanine ammonia-lyase (PAL) activity as well as essential oil composition of in vitro cultures and field grown rhizomes of Zingiber montanum. RESULTS: Methyl jasmonate-treated cell cultures showed the highest total phenolic content and steroid content of 22.23 mg gallic acid equivalent/g dry weight (DW) and 41.67 mg/g DW, respectively. Callus cultures exhibited the highest tannin content (39.53 mg tannic acid equivalent/g DW) and strongest antioxidant activity (91.05% inhibition of 2,2-Diphenyl-1-picrylhydrazyl or DPPH). The highest saponin (81.76 mg/g DW) and alkaloid (113.97 mg/g DW) contents were obtained in in vitro microrhizomes induced on Murashige and Skoog (MS) medium supplemented with 6% sucrose and 5 mg/l 6-Benzylaminopurine (BAP), and MS medium supplemented with 7% sucrose, respectively. The essential oil content varied in cell cultures and microrhizomes and mainly consisted of fatty acid esters, which are precursors of many secondary metabolites. Trace amounts of terpinen-4-ol (0.21 and 0.27 mg/g) and zerumbone (0.0107 mg/g) were also detected in the in vitro microrhizomes. CONCLUSION: The results obtained indicate the potential of in vitro cultures of Z. montanum for the production of secondary metabolites.

Jishengella zingiberis sp. nov., isolated from root tissue of Zingiber montanum.

A novel actinomycete, strain PLAI 1-1T, which formed spiny single spore directly on substrate mycelium was isolated from root tissue of Zingiber montanum. The isolate contained meso-diaminopimelic acid and 3-hydroxydiaminopimelic acid in the cell-wall peptidoglycan. The acyl type of the cell-wall muramic acid was glycolyl. The whole-cell sugars of strain PLAI 1-1T were glucose, arabinose, xylose, ribose and a trace amount of mannose. The membrane phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol. The major menaquinone was MK-9 (H4). The main cellular fatty acids were iso-C15 : 0 and C17 : 1ω8c. The G+C content of the genomic DNA was 70.6 mol%. 16S rRNA gene sequence analysis revealed that strain PLAI 1-1T was a member of the genus Jishengella and had the highest 16S rRNA gene sequence similarity to Jishengella endophytica DSM 45430T (99.2 %). Based on the data of physiological and biochemical tests, including the result of DNA-DNA hybridization, strain PLAI 1-1T represents a novel species of the genus Jishengella, for which the name Jishengellazingiberis sp. nov. is proposed. The type strain is PLAI 1-1T (=TBRC 7644T=NBRC 113144T).

Chemical composition and antibacterial, antifungal, allelopathic and acetylcholinesterase inhibitory activities of cassumunar-ginger.

BACKGROUND: Zingiber montanum (J.Koenig) Link ex A.Dietr. (Zingiberaceae), commonly known as cassumunar-ginger, is a folk remedy for the treatment of inflammations, sprains, rheumatism and asthma. The aim of the present study was to assess the chemical composition, and antibacterial, antifungal, allelopathic and acetylcholinesterase inhibitory activities of the essential oil of Z. montanum originating from India. RESULTS: The hydrodistilled essential oil of Z. montanum rhizome was analyzed using gas chromatography-flame ionization detection and gas chromatography-mass spectrometry. A total of 49 constituents, forming 98.7-99.9% of the total oil compositions, was identified. The essential oil was characterized by higher amount of monoterpene hydrocarbons (32.6-43.5%), phenylbutanoids (27.5-41.2%) and oxygenated monoterpenes (11.4-34.1%). Major constituents of the oil were sabinene (13.5-38.0%), (E)-1-(3',4'-dimethoxyphenyl)buta-1,3-diene (DMPBD) (20.6-35.3%), terpinen-4-ol (9.0-31.3%), γ-terpinene (1.1-4.8%) and ß-phellandrene (1.0-4.4%). The oil was evaluated against eight pathogenic bacteria and two fungal strains. It exhibited low to good antibacterial activity (minimum inhibitory concentration: 125-500 µg mL-1 ) and moderate antifungal activity (250 µg mL-1 ) against the tested strains. The oil reduced germination (69.8%) and inhibited the root and shoot growth of lettuce significantly (LD50 : 3.58 µL plate-1 ). However, it did not demonstrate acetylcholinesterase inhibitory activity up to a concentration of 10 mg mL-1 . CONCLUSIONS: The essential oil of Z. montanum can be used as a potential source of DMPBD, terpinen-4-ol and sabinene for pharmaceutical products. The results of the present study add significant information to the pharmacological activity of Z. montanum native to India. © 2017 Society of Chemical Industry.

A novel diphenylbutenoid-type compound from the rhizomes of Zingiber montanum (J.Koenig) Link ex A.Dietr. (Zingiberaceae).

From the EtOAc-soluble extract of the rhizomes of Zingiber montanum (J.Koenig) Link ex A.Dietr., a novel diphenylbutenoid, montadinin A (1) and a previously unreported phenylbutenoid compound, 1-(3,4-dimethoxyphenyl)but-3-en-2-ol (7), in natural source were isolated. Additionally, seven known phenylbutenoids were also identified. The structures of all compounds were elucidated through NMR spectroscopic interpretation. Compounds cis-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene (2), cis-4-[(E)-3,4-dimethoxystyryl]-3-(2,4,5-trimethoxyphenyl)cyclohex-1-ene (3), trans-3-(3,4,-dimethoxyphenyl)-4-[(E)-2,4,5-trimethoxystyryl]cyclohex-1-ene (5), and cis-3-(3,4-dimethoxyphenyl)-4-[(Z)-2,4,5-trimethoxylstyryl]cyclohex-1-ene (6) showed weak cytotoxicity against HepG2 cells with IC50 values of 122.9, 127.3, 257.5, and 168.5 µM, respectively.

Potential bangle (Zingiber montanum J.König) rhizome extract as a supplement to prevent and reduce symptoms of Covid-19.

The morbidity and mortality rates due to Covid-19 are increasing day by day, to overcome this, we urgently need a better treatment strategy, therefore various ways and strategies for this must be pursued. The purpose of the present review is to explain that the rhizome of bangle (Zingiber montanum) has great potential to increase antibodies and reduce symptoms of acute respiratory distress syndrome (ARDS), which also seems suitable for treating Covid-19. Method: This review is looking for the results of scientific research from various sources, regarding the efficacy of bangle (Zingiber montanum) rhizome which is strongly suspected to be able to prevent, and reduce the symptoms that occur in COVID-19. The results showed that the bangle rhizome extract had activity as immunomodulatory, antiviral and reduced symptoms such as what happened in COVID-19. Conclusion: Bangle rhizome extract has dozens of nutritious substances and has multifunctional activities, and it can be postulated that among the benefits of bangle rhizome extract it is able to prevent and reduce symptoms that occur in Covid-19, and preclinical studies and clinical studies are needed to prove this postulate.

Antioxidant and Anti-Inflammatory Activities of Zingiber montanum Oil in HepG2 Cells and Lipopolysaccharide-Stimulated RAW 264.7 Macrophages.

Improvement of antioxidant and anti-inflammatory functions is believed to be an effective strategy for protection against various diseases such as cancer, aging, and neurodegenerative disease. This study focused on investigating antioxidant and anti-inflammatory abilities of Zingiber montanum oil (ZMO) extracted by the supercritical CO2 fluid system in HepG2 cells and lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Ten predominant constituents of ZMO were identified, in which triquinacene, 1,4-bis (methoxy), terpinen-4-ol, triquinacene, 1,4,7-tris (methoxy), α-terpinene, sabinene hydrate, and (E and Z)-1-(3,4-dimethoxyphenyl)butadiene account for 86.47%. ZMO exhibited anti-inflammatory capacity by inhibiting the formation of pro-inflammatory markers such as nitric oxide, inducible nitric oxide synthase, cyclooxygenase-2, interleukin (IL)-1ß, IL-6, and monocyte chemoattractant protein-1 in LPS-treated macrophages. The LPS-induced stimulation of nuclear factor-kappa B, signal transducer and activator of transcription 3 (Stat3) and mitogen-activated protein kinase (MAPK) pathways as evident from increased phosphorylation of IKKα/ß, IκBα, p65, Stat3, ERK, JNK, and p38 MAPK was also suppressed by ZMO pretreatment. Further, ZMO enhanced the expression of nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase-1 (HO-1), and concurrently, reduced intracellular reactive oxygen species accumulation in LPS-treated RAW 264.7 cells. In addition, ZMO treatment markedly upregulated the expression of Nrf2 as well as its target genes, HO-1 and NAD(P)H:quinone oxidoreductase 1 in HepG2 cells. These data propose that ZMO may be a potent candidate for prevention and/or treatment of inflammatory and oxidative conditions.

The efficacy of honey and a Thai Herbal Oil preparation in the treatment of pressure ulcers based on Thai traditional medicine wound diagnosis versus standard practice: An open-label randomized controlled trial.

BACKGROUND: Scientific support for Thai traditional medicine (TTM) practice is warranted for reintroduction into modern healthcare systems. A promising TTM practice for treatment of pressure ulcers was selected to conduct a clinical trial. This study aimed to evaluate the efficacy of the TTM practice for the treatment of pressure ulcers using honey or a Thai Herbal Oil preparation (THO) based on the TTM wound diagnosis comparing with the standard practice. METHODS: The study design was an open-label randomized controlled trial. Sixty-six participants, with pressure ulcers at least stage II-IV or unstageable, were allocated to two groups via minimization. A TTM practice group received honey or THO depending on the TTM diagnosis via the Thai Traditional Medicine Pressure Ulcer Assessment Tool (TTM-PUAT). A standard practice group received advanced dressings, including hydrogel, alginate, silver-impregnated, or hydrocolloid dressings. The primary outcome was the Pressure Ulcer Scale for Healing (PUSH). RESULTS: Both TTM practice and standard practice showed a significant reduction in PUSH scores after treatments. However, there was no significant difference in PUSH score reduction between the groups. The mean PUSH score reduction over the 6-week period was 2.58 ± 3.38 (95% CI 1.34-3.82) in the TTM practice group and 3.24 ± 3.49 (95% CI 1.91-4.57) in the standard practice group (p = 0.284). The TTM practice and standard practice accelerated pressure ulcer healing without statistically significant difference between the practices, during 6 weeks in a home-based care setting. This finding supported the TTM practice as an alternative treatment for pressure ulcer.

Acute effects of essential oil blend containing phlai oil on mood among healthy male volunteers: Randomized controlled trial.

Background Essential oil blend (ESOB) containing Zingiber montanum or Phlai and Zingiber tenuiscapus oils as the major components has high levels of terpenes such as sabinene, terpinene, terpinolene, and terpine-4-ol. Phlai is well known for its analgesic property, but its effects on mood remain unclear. This study aimed to investigate the acute effects of inhaling the ESOB containing phlai essential oils on mood and stress among healthy volunteers. Methods A prospective, randomized, double-blinded, controlled trial pilot study was conducted among 38 healthy male volunteers; 19 in the ESOB containing phlai oil group and 19 in the control group, which used olive oil. One single inhalation was provided continuously for 15 min via hot steam. Mood status was assessed pre- and post-intervention using the Visual Analog Scales for Mood, Anxiety, Stress and Alertness (MASA). Rorschach tests, in a modified analysis, were also employed. One-way ANCOVA was used to determine statistical differences of the mood test results between ESOB containing phlai oil and olive oil. Results Mean age of the participants was 22.11 (SD 2.1) years. Both ESOB containing phlai oil and olive oil increased the post-test scores of positive mood and reduced the scores of negative mood. However, no difference was observed regarding the mood test scores between groups, except concerning the Rorschach stress test (F (1,38)=6.486, p=0.015). The variance of change, explained by the model (R square), was 32.8%, while the adjusted R-squared was 0.290. Conclusions Single inhalation aromatherapy using ESOB containing phlai oil or olive oil showed an immediate improvement of mood states among healthy young male volunteers by easing the negative emotions and enhancing positive emotions, especially feelings of pleasure and contentment. Natural blend of phlai essential oils was more efficacious than olive oil in reducing emotional stress.

Anti-hypercholesterolemic effect of Zingiber montanum extract.

Background: Hypercholesterolemia, high cholesterol levels in the blood, can contribute to many forms of disease, most notably cardiovascular disease. Anti-hypercholesterolemic agents generally used for those conditions have several side effects for patients. Zingiber montanum , known locally as "bangle", belongs to the family Zingiberaceae and is a potential plants for alternative anti-hypercholesterolemic agents. This plant, from East Kalimantan, is used in traditional medicine for health problems caused by high cholesterol levels. The aim of this research was to find alternatives to anti-hypercholesterolemic agents, especially from natural sources. Methods: This study was an experimental study using 30 Wistar male white rats. Subjects were randomly divided into 6 groups (n=5): (1) normal control group; (2) high fat diet control group; (3) high fat diet with simvastatin; (4-6) high fat diet with Zingiber montanum extracts 100, 200, and 400 mg/kg. After 4 weeks of treatment, blood was collected from all groups, and plasma concentrations of triglycerides, total cholesterol, high density lipoproteins (HDL), and low density lipoproteins (LDL) were measured. Results: The results showed significant differences in total cholesterol (p=0.000), LDL (p=0.000) and triglycerides (p=0.001) in the high-fat diet group with Z. montanum extract, as compared to the high-fat diet control. Meanwhile, there were no significant differences in HDL levels (p=0.830) between the high-fat diet group and other groups. The results also showed significant differences in total cholesterol and LDLs for rats treated with Z. montanum extract, 100 mg/kg (p=0.000), 200 mg/kg (p=0.000), and 400 mg/kg (p=0.000) compared to the high-fat diet group. The result of Z. montanum 400 mg/kg also showed a significant reduction, not only for total cholesterol and LDLs, but also for triglycerides (p=0.030). Conclusion: It could be concluded that Z. montanum extracts have the potency to be further developed as a new natural source of the anti-hypercholesterolemic agents.

Purification, biochemical characterization and antioxidant property of ZCPG, a cysteine protease from Zingiber montanum rhizome.

Zingiber montanum cysteine protease glycoprotein (ZCPG) was purified to homogeneity by DEAE- cellulose and Sephadex G50 resulting in sixteen fold purification and total activity of 39.4U/mg. ZCPG presented a prominent single peak in HPLC chromatogram with an estimated molecular weight of 48kDa on native PAGE. SDS-PAGE gave two subunits of ∼24.3 and ∼24.6kDa showing its heterodimeric form. Protein sequencing was studied by MALDI-TOF MS/MS. Isoelectrofocusing exhibited two isoforms with pI values of 4.8 and 5.1. Analysis of the total carbohydrate by GC-MS/MS showed the presence of glucose, mannose, fucose and xylose. The pH and temperature optimum were 9 and 60°C respectively while Km and Vmax values were 0.5±0.03µg and 13.73±2.07U/ml respectively. ZCPG was strongly inhibited by NEM indicating the cysteine-type. Substrates such as casein, azocasein, gelatin, BSA and haemoglobin showed high relative activity. Metal ions of CuCl2, CoCl2, HgCl2 and ZnCl2 showed partial inhibition at 1mM concentration. Furthermore, ZCPG exhibited promising antioxidant activity in biochemical systems as well as THP-1 cells. These findings suggested, ZCPG with significant antioxidant activity might have potential applications in therapeutic and food industry.

Chemical composition and larvicidal activity of essential oils from Zingiber montanum (J. Koenig) Link ex. A. Dietr. against three mosquito vectors / Composición química y actividad larvicida de aceites esenciales de Zingiber montanum (J. Koenig) Link ex. A. Dietr. contra tres vectores de mosquitos

The chemical composition and larvicidal activity of essential oils derived from the leaves and rhizomes of Zingiber montanum (J. Koenig) Link ex. A. Dietr. were reported. The main compounds in the leaf oil were ß-pinene (13.8%), ß-phellandrene (11.3%) and α-pinene (7.3%) while the rhizome oil was dominated by sabinene (41.1%), terpinen-4-ol (22.7%) and (E)-nerolidol (14.3%). The minimum lethal concentration (larvicidal activity) LC50of the rhizome oil at 24 h against Aedes albopictus was 35.17 µg/mL, while LC50 values of 32.20 µg/mL and 31.12 µg/mL were obtained against Aedes aegypti and Culex quinquefasciatus respectively. At 48 h the oil displayed larvicidal action with LC50 values of 23.18 µg/mL, 25.58 µg/mL and 18.99 µg/mL respectively towards Ae. albopictus, Ae. Aegyptiand Cx. quinquefasciatus. The leaf oil did not exhibit significant mortality and larvicidal action. The results indicate the potential of rhizome essential oil of Z. montanumas a source of larvicidal agent.

En el presente trabajo se reportan la composición química y actividad larvicida de los aceites esenciales obtenidos de hojas y rizomas de Zingiber montanum (J. Koenig) Link ex. A. Dietr. Los principales compuestos en el aceite de hojas fueron ß-pineno (13.8%), ß-felandrene (11.3%) y α-pineno (7.3%); mientras que los más abundantes en el aceite de rizomas fueron sabineno (41.1%), terpinen-4-ol (22.7%) y (E)-nerolidol (14.3%). La concentración letal mínima (actividad larvicida) LC50 del aceite de riomas ante Aedes albopictus fue 35.17 µg/mL, mientras que los valores de LC50 de 32.20 µg/mL y 31.12 µg/mL fueron obtenidos ante Aedes aegyptiy Culex quinquefasciatus respectivamente. A las 48 horas, el aceite mostró acción larvicida con valores de LC50 de 23.18 µg/mL, 25.58 µg/mL y 18.99 µg/mL respectivamente, ante Ae. albopictus, Ae. Aegyptiand Cx. quinquefasciatus. El aceite de hojas no mostró mortalidad ni acción larvicida significativa. Los resultados indican el potencial del aceite esencial de rizomas de Z. montanum como una fuente de agentes larvicidas.