Significant production of vanillin and in vitro amplification of ech gene in local bacterial isolates

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.

Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.

Significant production of vanillin and in vitro amplification of ech gene in local bacterial isolates / Produção significativa de vanilina e amplificação in vitro do gene ech em isolados bacterianos locais

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.

Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.

Experimental evaluation of anti-inflammatory effect of ethanolic extract of Vanilla planifolia seeds in Wistar rats

Background: Inflammation is a protective response of the body to the harmful stimuli. Inflammation can be either acute or chronic, always associated pain, redness and loss of function. Vanilla planifolia (VP) is the aromatic plant, as per literature, it has anti-inflammatory activity, which has not been tested as per modern medicinal parameters. Non-steroidal anti-inflammatory drugs are commonly used for the treatment but have many adverse effects such as gastritis, hepatitis etc. Therefore, there is always a search for new safe drug. Aim and Objectives: The aim of the study is to evaluate the anti-inflammatory activity of VP seeds in acute and chronic animal model of inflammation. Materials and Methods: Rats weighing 150–200 g of either sex were included in the study. Acute Anti-inflammatory activity tested with carrageenan-induced paw edema and chronic with cotton pellet-induced granuloma model. Animals were divided into five groups – Gr-I Control, Gr-II Vehicle control, Gr-III Diclofenac sodium, Gr-IV (VPLD), Gr-V (VPHD). Drug treatment was given 1 h before carrageenan injection. Paw volume measured at different time interval with plethysmometer. In chronic model, drug treatment was given for 7 days after pellet implantation. On 8th day, pellets removed and dried in oven. Weight of wet and dry pellets from all the groups compared with vehicle control. Data obtained was analyzed with Graph pad prism 6. Results: Reduction in paw volume started in all drug treated groups after 1 h of treatment. Paw volume was significantly reduced (P < 0.001) in group III, IV & V in 5 h. reached to near normal. In chronic model, VPLD showed decrease in wet pellet (P < 0.01) and dry pellet weight (P < 0.05) significantly. VPHD was more effective in reducing wet pellet (P < 0.001) and dry Pellet weight (P < 0.01). Similar results were seen on the left side. Conclusion: VP seed extract showed promising anti-inflammatory effect in both models of inflammation.

Research progress on antidepressant substances and their mechanisms in Gastrodia elata / 中草药

Gastrodia elata is a valuable traditional Chinese medicine with many pharmacological activities such as calming, anti-depression, anti-anxiety, anti-oxidation, anti-virus, and anti-tumor. In this paper, the research progress of the pharmacodynamic basis and antidepressant mechanism of anti-depression effect of G. elata is summarized. And combined with the current clinical application of G. elata anti-depression, a reference is provided for the further research and development of G. elata.

Fixatives Increase the Efficacy of Gel Formulations containing Piper aduncum Linnaeus (Piperales: Piperaceae) Essential Oil as Repellent / Jurnal Sains Kesihatan Malaysia

@#Piper aduncum essential oil exhibit repellency activity and has a potential to be use as an alternative for synthetic repellent such as N,N-diethyl-3- methylbenzamide, (DEET). However, the volatility properties of the essential oil decrease their persistence as a topical repellent. Study has shown that formulation of the essential oil with some fixatives may increase their effectiveness. Therefore, this study was conduct to evaluate the effectiveness of gel formulation containing P. aduncum essential oil with two fixative; vanillin and paraffin oil. Gel formulations containing P. aduncum essential oil with 5% and 10% vanillin and 6% paraffin oil was prepared and tested against Aedes aegypti in laboratory using Standards and Industrial Research Institute of Malaysia (SIRIM) bioassay method. After 240 minute post-application, formulation containing 5% and 10% vanillin was able to provide >70% repellency percentage against mosquito while formulation containing 6% paraffin oil gives <30% repellency percentage. As conclusion, P. aduncum based repellent gel containing 5% and 10% vanillin was able to prolong the effect of P. aduncum essential oil as repellent against dengue vector in laboratory.

Improvement of inhibitors tolerance of Saccharomyces cerevisiae by overexpressing of long chain sphingoid kinases encoding gene LCB4 / 生物工程学报

By-products released from pretreatment process of lignocellulose seriously hinder the development of cellulosic fuel ethanol. Therefore, the great way to increase the efficiency of cellulosic ethanol production is improvement of Saccharomyces cerevisiae tolerance to these inhibitors. In this work, the effects of LCB4 gene overexpression on cell growth and ethanol fermentation in S. cerevisiae S288C under acetic acid, furfural and vanillin stresses were studied. Compared to the control strain S288C-HO, the recombinant strain S288C-LCB4 grew better on YPD solid medium containing 10 g/L acetic acid, 1.5 g/L furfural and 1 g/L vanillin. Ethanol yields of recombinant strain S288C-LCB4 were 0.85 g/(L·h), 0.76 g/(L·h) and 1.12 g/(L·h) when 10 g/L acetic acid, 3 g/L furfural and 2 g/L vanillin were supplemented into the fermentation medium respectively, which increased by 34.9%, 85.4% and 330.8% than the control strain S288C-HO. Meanwhile, ethanol fermentation time was reduced by 30 h and 44 h under furfural and vanillin stresses respectively. Further metabolites analysis in fermentation broth showed that the recombinant strain produced more protective compounds, such as glycerol, trehalose and succinic acid, than the control strain, which could be the reason for enhancing strain tolerance to these inhibitors from pretreatment process of lignocellulose. The results indicated that overexpression of LCB4 gene could significantly improve ethanol fermentation in S. cerevisiae S288C under acetic acid, furfural and vanillin stresses.

Potassium Bromate-induced Changes in the Adult Mouse Cerebellum Are Ameliorated by Vanillin / 生物医学与环境科学（英文）

OBJECTIVE@#The current study aimed to elucidate the effect of vanillin on behavioral changes, oxidative stress, and histopathological changes induced by potassium bromate (KBrO3), an environmental pollutant, in the cerebellum of adult mice.@*METHODS@#The animals were divided into four groups: group 1 served as a control, group 2 received KBrO3, group 3 received KBrO3 and vanillin, and group 4 received only vanillin. We then measured behavioral changes, oxidative stress, and molecular and histological changes in the cerebellum.@*RESULTS@#We observed significant behavioral changes in KBrO3-exposed mice. When investigating redox homeostasis in the cerebellum, we found that mice treated with KBrO3 had increased lipid peroxidation and protein oxidation in the cerebellum. These effects were accompanied by decreased Na+-K+ and Mg2+ ATPase activity and antioxidant enzyme gene expression when compared to the control group. Additionally, there was a significant increase in cytokine gene expression in KBrO3-treated mice. Microscopy revealed that KBrO3 intoxication resulted in numerous degenerative changes in the cerebellum that were substantially ameliorated by vanillin supplementation. Co-administration of vanillin blocked the biochemical and molecular anomalies induced by KBrO3.@*CONCLUSION@#Our results demonstrate that vanillin is a potential therapeutic agent for oxidative stress associated with neurodegenerative diseases.

Vanillin Schiff bases: Molecular interactions in methanol and THF solutions / Bases de Schiff de vainillina: interacciones intermoleculares en soluciones de metanol y THF

SUMMARY Density, ultrasonic velocity and viscosity of some vanillin Schiff bases derivatives have been studied in methanol and tetrahydrofuran (THF) at 308.15 K. From the experimental data, various acoustical parameters such as isentropic compressibility (κ s), Rao's molar sound function (R m ), Van der Waals constant (b), relaxation strength (r), intermolecular free length (L f ), apparent molar compressibility, etc. have been evaluated, which helps in understanding the molecular interactions occurring in these solutions.

RESUMEN En este trabajo se estudiaron la densidad, la velocidad ultrasónica y la viscosidad de soluciones de algunas bases de Schiff derivadas de la vainillina en metanol y tetrahidrofurano (THF) a 308,15 K. A partir de los datos experimentales, se evaluaron diversos parámetros acústicos, como la compresibilidad isentrópica (k s ), la función acústica molar de Rao (R m ), la constante de Van der Waals (b), la fuerza de relajación (r), la longitud intermolecular libre (L f ), la compresibilidad molar aparente, etc., todo lo cual ayuda a comprender las interacciones moleculares que ocurren en estas soluciones.

Evaluation of enzymatic activity and phenolic compounds during microbial transformation of curcuma longa to vanillin

Aims: Evaluation of enzymes responsible for microbial transformation of Curcuma longa to vanillin and quantification of phenolic compounds present during the process. Methodology and results: Yeast strains isolated from fermented beverages using curcumin-yeast extract agar were screened for their ability to transform curcumin to vanillin by evaluating ferulic acid esterase, ferulic acid decarboxylase, and vanillin dehydrogenase activities in growing cultures and cell-free supernatant. Based on bioconversion and enzymatic capabilities of the yeasts, Pichia angusta BT21 was selected for microbial transformation of Curcuma longa to vanillin using submerged fermentation technique under stationary mode. The metabolites were extracted from the transformation medium using solid-phase extraction technique and analyzed by thin-layer chromatography and gas chromatography-flame ionization detector. Phenolic compounds obtained in this research comprised of flavonoids (flavanols, flavones, isoflavones and flavonones), phenolic aldehyde (vanillin, vanillic acid, isoeugenol), simple phenolic (phenol) and phenolic acids (the hydroxybenzoic and the hydroxycinnamic acids) respectively. The research is quite profitable from an industrial point of view, considering the commercial price of vanillin and low cost and availability of C. longa which will eventually reduce the cost of industrial production of vanillin and increase supply. Conclusion, significance and impact of study: Bioprocessing of C. longa by submerged fermentation technique under stationary mode reduces vanillin dehydrogenase activity of the Pichia angusta BT21, thus preventing the degradation of vanillin which subsequently leads to increase in vanillin concentration.

Chemical Constituents of Ethyl Acetate Fraction from Cynodon dactylon / 中国药学杂志

OBJECTIVE: To isolate and identify the chemical constituents from Cynodon dactylon. METHODS: The chemical constituents of the alcohol extract of C. dactylon. were isolated and purified by various chromatography methods, and their structures were identified by physical and chemical properties and spectral data. RESULTS: Fourteen compounds were isolated from C. dactylon and their structures were examined by physicochemical characteristics and spectral data and identified as vanillin (1), arctigenin (2), maltol (3), (+)-dehydrovomifoliol (4), (3R, 6R, 7E)-3-hydroxy-4, 7-megastigmadien-9-one (5), (-)-loliolide (6), zhebeiresinol (7), 1-indole-3-carboxaldehyde (8), ferulic acid (9), matairesinol (10), pinoresinol (11), ethyl caffeate (12), traxillagenin (13), and impecylenolide (14). CONCLUSION: Compounds 1-14 are isolated from C. dactylon for the first time.

Golgi dispersal in irradiated cells and the protective effect of vanillin derivatives / 军事医学

Objective To determine the Golgi dispersal in radiation damaged cells and the protective effect of vanillin derivatives.Methods Immunofluorescence, cell cycle analysis of flow-cytometry,Western blot,and clone formation were used.Results Immunofluorescence observation showed that the Golgi dispersal caused by 2 Gy 60 Coγ-ray was significantly increased in a dose-dependent manner in the range of 4-10 Gy as was demonstrated by the fact that the Golgi area was significantly increased. When the irradiated cells were treated with the radioprotective agent VND3207, a vanillin derivative,the Golgi dispersal induced by radiation was significantly reduced.The radiation-induced Golgi dispersal was also displayed in a pattern of time-course after irradiation in the HeLa cells, and persisted at least to 36 h post-irradiation. Cell cycle test results indicated that the Golgi dispersal was not associated with the G2/M arrest triggered by radiation-induced DNA damage response.VND3207 could promote cell survival by plate colony formation assay.Conclusion The Golgi dispersal can be caused byγ-ray irradiation in a dose-and time-dependent manner, and VND3207 can provide a good protection against radiation injury associated with inhibited Golgi dispersal.

Chemical constituents from flowers of Gardenia jasminoides / 中草药

Objective: To study the chemical constituents from the flowers of Gardenia jasminoides. Methods: The compounds were isolated by extensive chromatographic methods, including macroporous adsorption resin, silica gel column chromatography, and HPLC method. Their structures were elucidated on the basis of physiochemical properities and spectroscopic data. Results: Ten compounds were obtained and identified as kaempferol 3-O-β-D-glucopyranoside (1), kaempferol 3-O-β-D-galactopyranoside (2), kaempferol 3-O- robinobioside (3), kaempferol 3-O-rutinoside (4), corchoionoside C (5), (E)-p-hydroxycinnamic acid methyl ester (6), vanillin (7), caffeic acid (8), protocatechuic acid (9), and β-sitosterol (10). Among them, compounds 1-4 belong to flavonoid glucosides. Conclusion: Compounds 2, 3, and 5-7 are first isolated from the plants of Gardenia Ellis and compounds 1, 4, 8 and 9 are first isolated from the flowers of G. jasminoides.

Chemical constituents from rhizomes of Curcuma longa / 中草药

Objective: To study the chemical constituents in the rhizomes of Curcuma longa. Methods: A variety of silica gel column chromatography, Sephadex LH-20 gel column chromatography, and HPLC methods were used for the separation and purification of chemical composition. Their structures were established on the basis of physicochemical property and spectral data. Results: Thirteen compounds were obtained and identified as 5-hydroxyl-bisabolon-9-one (1), cyclocurcumin (2), demethoxyl cyclocurcumin (3), isodemethoxyl cyclocurcumin (4), curcumin (5), demethoxyl curcumin (6), methyl ferulate (7), vanillin (8), 4-hydroxyl benzoic aldehyde (9), 4-(4-hydroxyl phenyl)-2-butanone (10), 4-(4-hydroxyl-3-methoxyl phenyl)-2-butanone (11), 4-(4-hydroxyl phenyl)-3- buten-2-one (12), and 4-(4-hydroxyl-3-methoxyl phenyl)-3-buten-2-one (13). Conclusion: Compound 1 is obtained as a new compound named 5-hydroxyl-bisabolon-9-one without reported in literature until now and compound 4 is first isolated from this plant.

Some numerical analyses on the solubility of vanillin in Carbitol® + water solvent mixtures / Algunos análisis numéricos sobre la solubilidad de la vainillina en mezclas cosolventes Carbitol® + agua / Alguns análises numéricas da solubilidade de vanilina em misturas de solventes carbitol® + água

In this communication some reported solubility values of vanillin (component 3) in 2-(2-ethoxyethoxy)ethanol (Carbitol®, component 1) + water (component 2) mixtures at five temperatures from 298.15 to 318.15 K were correlated with the Jouyban-Acree model combined with van't Hoff or Apelblat equations, obtaining models in second degree regarding the mixtures compositions. Mean percentage deviations were near to 6.0%. On the other hand, by means of the inverse Kirkwood-Buff integrals it was demonstrated that vanillin is preferentially solvated by water in water-rich mixtures (with a minimum δx1,3 value in the mixture x1 = 0.05, i.e. -4.29 x 10-2) but preferentially solvated by the cosolvent in mixtures with compositions 0.12 < x1 < 1.00 (with a maximum δx1,3 value equal to 3.61 x 10-2 in the mixture x1 = 0.25). It is conjectural that hydrophobic hydration plays a relevant role in the first case, whereas, in the second case, vanillin would be acting as Lewis acid with Carbitol®.

Nesta pesquisa alguns valores de solubilidade da vanilina (componente 3) em misturas 2-(2-etoxietoxi)etanol (Carbitol®, componente 1) + água (componente 2) em várias temperaturas (298,15-318,15 K) foram correlacionados com o modelo Jouyban-Acree combinado com as equações do van't Hoff ou Apelblat obtendo modelos de ordem dois. Os desvios médios percentuais foram cercanos a 6,0%. Por outro lado, por meio das integrais inversas do Kirkwood-Buff demonstrou-se que a vanilina é preferencialmente solvatada pela água em misturas ricas em agua (com um valor mínimo de δx1,3 igual a -4,29 x 10-2 obtido na mistura de composição x1 = 0,05), mas, preferencialmente, solvatada pelo cosolvente, em misturas com composições 0,12 < x1 < 1,00 (com um valor máximo de δx1,3 igual a 3,61 x 10-2 obtido na mistura de composição x1 = 0,25). É conjecturável que a hidratação hidrofóbica desempenha um papel relevante no primeiro caso, enquanto que, no segundo caso a vanilina está atuando como ácido de Lewis com moléculas do Carbitol®.

En esta comunicación se presenta la correlación de algunos valores de solubilidad de vainillina (componente 3) en mezclas 2-(2-etoxietoxi)etanol (Carbitol®, componente 1) + agua (componente 2) reportados previamente en la literatura a cinco temperaturas desde 298,15 hasta 313,15 K mediante el modelo de Jouyban-Acree combinado con las ecuaciones de van't Hoff y de Apelblat. En el análisis se obtuvieron modelos de segundo orden respecto a la composición de las mezclas disolventes. Las desviaciones porcentuales promedio fueron cercanas al 6,0%. Por otro lado, mediante las integrales inversas de Kirkwood-Buff se demostró que la vainillina es solvatada preferencialmente por el agua en mezclas ricas en agua (con un valor mínimo de δx1,3 igual a -4,29 x 10-2 obtenido en la mezcla de composición x1 = 0,05) pero preferencialmente solvatada por el cosolvente en mezclas con composiciones 0,12 < x1 < 1,00 (con un valor máximo de δx1,3 igual a 3,61 x 10-2 obtenido en la mezcla de composición x1 = 0,25). Se podría conjeturar que la hidratación hidrofóbica juega un papel relevante en el primer caso, mientras que en el segundo caso, la vainillina estaría actuando como ácido de Lewis frente a las moléculas de Carbitol®.

Effect of vanillin inhalation on brain derived neurotrophic factor in depressed model rats / 中华行为医学与脑科学杂志

Objective To evaluate the relieving effects of vanillin sinffing on depression-like behaviors in depressed rats and to explore the possible underlying mechanism.Methods Depression animal model established by chronic unpredictable medium intensity stress combined with isolation and destroy the olfactory bulb.The depressed rats were divided randomly into vanillin inhalation group,fluoxetine hydrochloride group,depression model group,olfactory bulbectomy with the vanillin inhalation treatment group and sham-operated group.Nervous behavioral changes had been observed at different time after the administration of 5 weeks.The concentration of brain derived neurotrophic factor(BDNF) in the brain homogenate and the positive expression of BDNF in hippocampus had been also measured.Results Two weeks after the intervention,the immobility time of vanillin group((12.78 ±7.50) s) was lower than that of the model group((57.33±32.16) s) (P＜0.05).The consumption of saccharose in vanillin group((52.88±25.18)g) was higher than that of model group((37.40±19.33) g) (P＜0.05).BDNF of the brain homogenate in vanillin group (0.54±0.13) was significantly increased compared with model group (0.36± 0.06) (P＜0.01).When compared with the OBX group (0.40±0.06),similar result was obtained.Immunohistochemistry and the average density of image analysis revealed that the expression of BNDF of hippocampal CA3 in vanillin group (0.40±0.03)was significantly increased compared with model group (0.25±0.04) and OBX group (0.28±0.03) (P＜0.01).Conclusion Vanillin inhalation significantly relieves depression-like behaviors in depression rats.The possible mechanism may increase hippocampal neurogenesis by raising brain derived neurotrophic factor in brain.

Isolation and cloning of an aryl-aldehyde dehydrogenase gene from the white-rot fungus Pycnoporus cinnabarinus strain MUCL 39533

Aims: The white rot fungus Pycnoporus cinnabarinus MUCL 39533 is able to reduce vanillic acid to vanillin. Reduction of vanillic acid to vanillin catalysed by the key enzyme aryl-aldehyde dehydrogenase has been reported. Here we report the isolation and cloning of aryl-aldehyde dehydrogenase from P. cinnabarinus strain MUCL 39533. Methodology and results: An aryl-aldehyde dehydrogenase gene (PcALDH) was isolated from P. cinnabarinus by producing a partial cDNA sequence fragment of an aryl-aldehyde dehydrogenase gene through PCR. Degenerate PCR primers were designed based on codons corresponding to conserved amino acid regions of aryl-aldehyde dehydrogenases of several fungi and bacteria. The full-length PcALDH cDNA was obtained through ReverseTranscription-Polymerase Chain Reaction (RT-PCR) and Rapid Amplification cDNA Ends (RACE) PCR. PcALDH cDNA comprises an open reading frame of 1,506 bp that encodes a protein of 501 amino acids. The PcALDH predicted protein showed the highest amino acid sequence identity (84%) to ALDH from Trametes versicolor. In silico analysis of PcALDH indicated that it belongs to the ALDH super-family and Class 3 ALDH. Conclusion, significance and impact study: PcALDH cDNA was successfully isolated and characterized. Important motifs identified from the highly conserved PcALDH protein indicated that it belongs to the aldehyde dehydrogenase superfamily. The cDNA clone will be used in expression studies to confirm the catalytic function of the enzyme.

Saponin Accumulation in Flower Buds of Panax notoginseng / 中草药·英文版

Objective: Panax notoginseng is an important Chinese medicinal plant. Saponin accumulation is higher in the flower buds than in the other parts of P. notoginseng. However, the flower bud compositions have not yet been quantified. The aim of this study is to investigate the formation and accumulation of saponins in the flower buds of P. notoginseng from different populations and at different growth years. Methods: Fourteen types of P. notoginseng with different growing durations and from different areas of Wenshan County, Yunnan Province were collected. We separated P. notoginseng individually into the flower buds, stems, leaves, and roots at the places where it has the highest saponin content. An efficient high-performance liquid chromatography (HPLC) method was developed for simultaneously quantifying two active saponins, ginsenoside Rb1 and ginsenoside Rb3, in the flower buds of P. notoginseng. The total saponin content was determined by using a quantitative vanillin-sulfuric acid colorimetric method. HPLC method was used to establish the fingerprints of 13 saponins and then quantify the composition in the whole plant of P. notoginseng. Results: The saponin contents of different parts differ significantly, and the total saponin content and those of Rb1 and Rb3 do not entirely correlated. The flower buds of P. notoginseng contain 27.79% of total saponins, which is the highest saponin content in the whole plant. Fingerprint result showed that different saponins were appeared in different parts of the plant, i.e. flower buds, stems, leaves, and roots. Conclusion: The saponin contents from the flower buds of P. notoginseng vary depending on the growth area and duration. The fingerprints show that the saponin contents and compositions vary depending on the part of P. notoginseng. These results are useful for the pharmacological evaluation and quality control of P. notoginseng.

Effect of vanillin via olfactory pathway on repetitive stereotyped behavior in autistic rats / 中华行为医学与脑科学杂志

Objective To investigate the effect of vanillin via olfactory pathway on repetitive stereotyped behavior in autistic rats and its possible mechanism.Methods The model of autism was established by the single intraperitoneal injection of 600 mg/kg sodium valproate on day 12.5 of pregnant rats.Twenty-four autism animals were adopted according to the eye opening time,body mass and behavioral test.They were randomly divided into control group and vanillin group with 12 in each group,while the saline group rats as normal control group and the blank group rats as normal blank group.The repetitive stereotyped behavioral were examined in four groups of rats and the concentration of 5-HT in the amygdala was determined by immunohistochemical stainning.Results Compared with the control group,the repetitive stereotyped behaviors of vanillin group decreased (stereotype time (113.31 ± 18.45) s vs (250.52± 13.25) s,P＜0.01) and the expression of 5-HT in amygdala increased (5-HT positive neuron number(3.42± 1.32) vs (1.32± 1.04),P＜0.05).Conclusions Vanillin via olfactory pathway can improve repetitive stereotyped behaviors of autism rats,which is associated with increased the expression of 5-HT in amygdala.

Determination of Vanillin in Dairy Products by HPLC / 中国药师

Objective:To establish an HPLC method to determine vanillin in 18 dairy products. Methods:The samples were sep-arated on a C18 column (250 mm × 4. 6 mm, 5 μm) with the mobile phase of methanol and water (35∶65) at the flow rate of 1. 0 ml ·min-1 . The detection wavelength was 275 nm. Results:The LOD ( limit of detection) of vanillin was 0. 45 μg·ml-1 . Excellent linearity was obtained within the concentration range of 1.990-99.500 μg·ml-1(r=1.000 0). The average recovery of vanillin in different dairy products with low, medium and high levels varied between 96. 26% and 100. 81% with RSD of 0. 15%-2. 22%. Con-clusion:The method is simple and reproducible, which can be applied in the rapid analysis of vanillin in dairy products.

Quality Standard for Yuhuang Sprays / 中国药师

Objective: To establish the quality standard for Yuhuang sprays. Methods: Berberine hydrochloride in Yuhuang sprays was qualitatively identified by TLC and determined by UV. The Vanillin-H2 SO4 method was employed for the determination of condensed tannins. Results:The spots obtained from the sample solutions showed the same color as those from the reference substance solution without interference from the negative sample. The calibration curve of berberiny hydrochloride was linear within the range of 1. 13-13. 60μg·ml-1(r=0. 999 9), and the average recovery was 99. 68% with RSD of 1. 53%(n=9). The calibration of catechin was linear within the range of 0. 03-0. 30 mg· ml-1(r=0. 999 3), and the average recovery was 99. 6% with RSD of 1. 76%(n=9). Conclusion:The method can be used in the quality control of Yuhuang sprays.