Shimadzu GC-2010 by Shimadzu - Scientific equipment at QuestPair.com

Shimadzu Shimadzu GC-2010

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Description

The Shimadzu GC-2010 is a gas chromatograph (GC) instrument that is used to separate, identify, and quantify the chemical components of a sample. It works by vaporizing the sample and injecting it into a long, narrow column that is filled with a stationary phase, such as a solid or liquid. As the vaporized sample moves through the column, the different components of the sample are separated based on their interactions with the stationary phase. The separated components then pass through a detector, which measures the amount of each component present in the sample. The Shimadzu GC-2010 is a high-performance GC instrument that is used in a variety of applications, including environmental analysis, food and beverage analysis, and petrochemical analysis. It features a high-sensitivity flame ionization detector (FID) and a temperature programmable injector and column, which allow for highly accurate and precise analysis of a wide range of samples.

This model was found at

3419 locations

The model is used in

78 countries

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About the Shimadzu GC-2010

The model Shimadzu GC-2010 was found in 3419 unique locations in 78 countries where it was mentioned from 1995 until recently. The Shimadzu GC-2010 is a gas chromatograph (GC) instrument that is used to separate, identify, and quantify the chemical components of a sample. It works by vaporizing the sample and injecting it into a long, narrow column that is filled with a stationary phase, such as a solid or liquid. As the vaporized sample moves through the column, the different components of the sample are separated based on their interactions with the stationary phase. The separated components then pass through a detector, which measures the amount of each component present in the sample. The Shimadzu GC-2010 is a high-performance GC instrument that is used in a variety of applications, including environmental analysis, food and beverage analysis, and petrochemical analysis. It features a high-sensitivity flame ionization detector (FID) and a temperature programmable injector and column, which allow for highly accurate and precise analysis of a wide range of samples. It is used by scientists in various research fields such as Organic Chemistry, General Medicine, Drug Discovery, Pharmaceutical Science, and General Chemistry. The model is also used in Molecular Medicine, Analytical Chemistry, Physical and Theoretical Chemistry, Biochemistry, Food Science, Chemistry (miscellaneous), Biotechnology, Molecular Biology, Plant Science, Microbiology, Applied Microbiology and Biotechnology, General Chemical Engineering, Pharmacology, Catalysis, Bioengineering, General Biochemistry, Genetics and Molecular Biology, Renewable Energy, Sustainability and the Environment, Animal Science and Zoology, Ecology, Evolution, Behavior and Systematics, Genetics, Environmental Engineering, Clinical Biochemistry, Microbiology (medical), Environmental Chemistry, and Waste Management and Disposal.

About Shimadzu

Shimadzu is a Japanese company founded as early as 1875 in Kyoto (Japan). The organisation manufactures a wide range of precision instruments for research & development and industrial applications. Their line of products includes liquid and gas chromatographs, spectrometers, elemental and surface characterisation systems, high-speed cameras and other analytical instruments. Shimadzu also provides instruments for testing purposes and applies their technology to product development and quality assurance applications.

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Organizations using this equipment

Research that uses the Shimadzu GC-2010

Optimization of cultivation conditions for biotechnological production of lipid by Rhodotorula kratochvilovae (syn, Rhodosporidium kratochvilovae) SY89 for biodiesel preparation

Abstract Rhodotorula kratochvilovae (syn, Rhodosporidium kratochvilovae) SY89, an oleaginous yeast, isolated from Ethiopian soil, was grown under nitrogen-limited media. The capacity this with respect to biomass production, lipid yield and lipid content was evaluated. The influence of inoculum size, carbon sources, variations in glucose concentration, nitrogen sources, C/N ratio, pH, temperature, agitation, and aeration rate and incubation period were investigated. Inoculum size of 10% v/v, glucose as a carbon source at 50 g/L glucose, 0.50 g/L yeast extract and 0.31 g/L (NH4)2SO4, C/N ratio of 120, pH 5.5, incubation temperature of 30 °C, 225 rpm, 0.2 as aeration ratio and 144 h of incubation were found to be optimum conditions for lipid production. Then the yeast was grown in a batch bioreactor by combining the different optimized parameters together. Under the optimized conditions, the yeast gave maximum biomass (15.34 ± 1.47 g/L), lipid yield (8.60 ± 0.81 g/L) and lipid content (56.06 ± 1.70%). The dominant fatty acids exhibited in order of their relative abundance (%w/w), were oleic, palmitic, linoleic, stearic, linolenic and palmitoleic acids. The concentration of saturated and monounsaturated fatty acids adds up 78.63 ± 2.19%. This suggests that this strain could be used as a good feedstock for biodiesel production.

Link: http://dx.doi.org/10.1007/s13205-017-0769-7

Artificial produced water as a medium to grow Chlorella sp. for biodiesel production

Increased interest in renewable, carbon-neutral energy sources makes processing biodiesel from microalgae has become the objective for many researchers and companies. Some kind of wastewater including municipal, industrial and agricultural wastewaters have been identified as alternate growth mediums. Produced water is the largest byproduct of the oil and natural gas extraction process which constitutes of high concentration of pollutants, such as dissolved nitrogen, phosphorus, dissolved organic carbon, heavy metal and monocyclic aromatic compound like BTEX. The purpose of this study is to identify Chlorella sp. potential for producing lipid in artificial produced water. Variations made in this study consist of 0%, 25%, 50%, 75% and 100% volume of artificial produced water to the control Walne medium. The highest specific growth rate and biomass productivity of Chlorella sp. achieved by culture grown in 25% wastewater with a value of 0.225 day−1 and 0.175 g L−1day−1, respectively. The highest lipid yield and productivity in mixed culture of artificial produced water and Walne medium achieved by culture in 25% artificial produced water with value of 0.231 and 40.48 mg.L−1.day−1. C16 and C18 fatty acids which dominated the lipids of Chlorella sp. in all culture variations indicated that the lipid of Chlorella sp. were suitable for producing high quality biodiesel.

Link: http://dx.doi.org/10.1051/e3sconf/202014802005

The Tomato Terpene Synthase Gene Family

Abstract Compounds of the terpenoid class play numerous roles in the interactions of plants with their environment, such as attracting pollinators and defending the plant against pests. We show here that the genome of cultivated tomato (Solanum lycopersicum) contains 44 terpene synthase (TPS) genes, including 29 that are functional or potentially functional. Of these 29 TPS genes, 26 were expressed in at least some organs or tissues of the plant. The enzymatic functions of eight of the TPS proteins were previously reported, and here we report the specific in vitro catalytic activity of 10 additional tomato terpene synthases. Many of the tomato TPS genes are found in clusters, notably on chromosomes 1, 2, 6, 8, and 10. All TPS family clades previously identified in angiosperms are also present in tomato. The largest clade of functional TPS genes found in tomato, with 12 members, is the TPS-a clade, and it appears to encode only sesquiterpene synthases, one of which is localized to the mitochondria, while the rest are likely cytosolic. A few additional sesquiterpene synthases are encoded by TPS-b clade genes. Some of the tomato sesquiterpene synthases use z,z-farnesyl diphosphate in vitro as well, or more efficiently than, the e,e-farnesyl diphosphate substrate. Genes encoding monoterpene synthases are also prevalent, and they fall into three clades: TPS-b, TPS-g, and TPS-e/f. With the exception of two enzymes involved in the synthesis of ent-kaurene, the precursor of gibberellins, no other tomato TPS genes could be demonstrated to encode diterpene synthases so far.

Link: http://dx.doi.org/10.1104/pp.111.179648

Highly sensitive VOC detectors using insect olfactory receptors reconstituted into lipid bilayers

A highly sensitive odorant sensor was developed by using insect olfactory receptors reconstituted into a lipid bilayer membrane.

Link: http://dx.doi.org/10.1126/sciadv.abd2013

Metabolic Modulation and Potential Biomarkers of the Prognosis Identification for Severe Aortic Stenosis after TAVR by a Metabolomics Study

Objectives. To investigate the metabolic profile in patients with aortic stenosis (AS) after transcatheter aortic valve replacement (TAVR) and explore the potential biomarkers to predict prognosis after TAVR based on metabolomics. Methods and Results. Fifty-nine consecutive AS patients were prospectively recruited. Blood samples from the ascending aorta, coronary sinus, and peripheral vein at before and after TAVR were collected, respectively. Liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry were performed to analyze the metabolic profile before and after TAVR. Influential metabolites were identified by integrating the univariate test, multivariate analysis, and weighted gene coexpression network analysis (WGCNA) algorithm. PLS-DA analysis revealed a significant extremely early (within 30 minutes after TAVR) alterations of metabolites in the ascending aorta, coronary sinus, and peripheral vein. The early (within 7 days after TAVR) changed metabolites in the peripheral vein were involved in purine metabolism, primary bile acid biosynthesis, glycerolipid metabolism, amino sugar and nucleotide sugar metabolism, one carbon pool by folate and alanine, and the aspartate and glutamate metabolism pathway. We used volcano plots to find that the cardiac-specific changed metabolites were enriched to the sphingolipid metabolism pathway after TAVR. Besides, WGCNA algorithm was performed to reveal that arginine and proline metabolites could reflect left ventricle regression to some extent. Conclusion. This is the first study to reveal systemic and cardiac metabolites changed significantly in patients with AS after TAVR. Some altered metabolites involved in the arginine and proline metabolism pathway in the peripheral vein could predict left ventricle regression, which merited further study.

Link: http://dx.doi.org/10.1155/2020/3946913

Estimation of Total Quercetin and Rutin Content in Malus domestica of Nepalese Origin by HPLC Method and Determination of Their Antioxidative Activity

Malus domestica (Apple) is one of the most widely cultivated cash crops of Nepal. Jumla and Mustang are two major pocket areas for the production of apple. Flavonoids including quercetin and rutin are potent antioxidants present in apples. This study was designed to quantify and compare the presence of quercetin and rutin in different plant parts (peel, leaf, and bark) among various cultivars of Malus domestica from two pocket zones of Nepal. A new HPLC-UV method was developed and validated for the quantification of quercetin and rutin. Polyphenols, flavonoids, and carbohydrate contents were determined by colorimetric methods. 2,2′-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay was carried out to measure in vitro antioxidative activity. Acid hydrolysis of each extract was carried out by the standard method to measure aglycone quercetin content after hydrolysis of its glycosides. The total rutin content ranged from 3.69 ± 1.34 to 374.50 ± 2.35 mg/100g dry extract weight. Before the acidic hydrolysis, the total quercetin content ranged from 2.96 ± 0.13 to 171.05 ± 0.95 mg/100g dry extract weight whereas its amount increased highly after the hydrolysis and it ranged from 80.84 ± 19.65 to 7445.32 ± 29.25 mg/100g dry extract weight. Total polyphenol content ranged from 19.48 ± 0.23 to 123.48 ± 1.84 µg gallic acid equivalent/mg of dry extract weight. Similarly, flavonoid content ranged from 2.21 ± 0.72 µg to 755.54 ± 1.91 µg quercetin equivalent/mg of dry extract weight. Total carbohydrate content ranged from 144.15 ± 3.73 to 484.65 ± 2.63 µg glucose equivalent per 0.5 mg dry extract weight. All the extracts showed the various degrees of antioxidant activity in a dose-dependent manner. Among them, stem bark of the Jonathan Jumla showed potent antioxidant activity with IC50 value of 13.003 µg/mL. The present study provides the information about variation of the phytochemical content among the different cultivars, parts, and geographic locations. Furthermore, it revealed that bark of Malus domestica cultivars had high quercetin and rutin content with high antioxidant activity.

Link: http://dx.doi.org/10.1155/2020/8853426

Analysis of Volatile Components of Auricularia auricula from Different Origins by GC-MS Combined with Electronic Nose

Auricularia auricula is a kind of nutrient-rich edible fungus, which has the reputation of “king of vegetarians.” In this paper, the electronic nose combined with GC-MS technology was used to analyze the volatile components of A. auricula in Heilongjiang, Jilin, Shanghai, and Sichuan provinces to investigate the differences and characteristics of A. auricula in different origins. The results showed that the electronic nose could obviously distinguish the samples from Jilin and Shanghai with a high degree of discrimination, while it was inappropriate to distinguish the samples from Heilongjiang and Sichuan Province. GC-MS was used to further analyze the volatile compounds in A. auricula qualitatively and quantitatively. The results showed that 98 volatile components were detected and 23 of them were common components, including alcohols, aldehydes, acids, esters, hydrocarbons, and other volatile components. The relative content of acetic acid and diethyl azodicarboxylate in A. auricula from the four origins was relatively high. According to the relative odor activity value (ROAV), it was found that the key compounds that caused the aroma difference between different origins were 1-octene-3-ol, cis-3-nonene-1-ol, (E)-2-octenal, (E)-2-nonenal, (E,E)-2,4-nonadienal, and 3-methyl butanal.

Link: http://dx.doi.org/10.1155/2020/8858093

Antibacterial Activity of Rosmarinus officinalis against Multidrug-Resistant Clinical Isolates and Meat-Borne Pathogens

Background. In developing countries, the prevalence of bacterial infections is quite rampant due to several factors such as the HIV/AIDS pandemic, lack of hygiene, overcrowding, and resistance to conventional antimicrobials. Hence the use of plant-based antimicrobial agents could provide a low-cost alternative therapy. Rosmarinus officinalis is reputed as a medicinal plant in Ethiopia; however, its antibacterial activity against many of the clinical isolates remains overlooked. Methods. Tender foliage of R. officinalis was collected and extracted in ethanol (EtOH) and evaluated for their antimicrobial activity against ten multidrug-resistant (MDR) clinical isolates, human type culture pathogens, and meat-borne bacterial isolates by employing agar well diffusion assay. Results. EtOH extract of R. officinalis efficiently subdued the growth of all tested MDR clinical isolates in varying degrees. Salmonella sp. and Staphylococcus aureus were found to be the most sensitive clinical isolates. Likewise, it efficiently repressed the growth of meat-borne pathogens, particularly, S. aureus and Salmonella sp. showing its potentiality to be used as a natural antibacterial agent in the meat processing industry. The mechanism of antibiosis of plant extract against meat-borne pathogens is inferred to be bactericidal. Chemical constituents of the crude plant extract were analysed by Gas Chromatography-Mass Spectroscopy (GC-MS), Fourier Transform Infrared (FT-IR), and UV-visible spectroscopy showing genkwanin (26%), camphor (13%), endo-borneol (13%), alpha-terpineol (12%), and hydroxyhydrocaffeic acid (13%) as the major compounds. Conclusion. Overall results of the present study conclude that R. officinalis could be an excellent source of antimicrobial agents for the management of drug-resistant bacteria as well as meat-borne pathogens.

Link: http://dx.doi.org/10.1155/2021/6677420

Pharmacokinetic Study of Safrole and Methyl Eugenol after Oral Administration of the Essential Oil Extracts of Asarum in Rats by GC-MS

Asarum is a traditional medicine and has been widely used as remedies for inflammatory diseases, toothache, headache, local anesthesia, and aphthous stomatitis in China, Japan, and Korea. Our previous research found that safrole and methyl eugenol were vital compounds that contribute to distinguish the different species and raw Asarum and its processed products apart. The pharmacokinetics of safrole and methyl eugenol after oral administration of Asarum extract has not been reported yet. In this study, a rapid and simple gas chromatography–mass spectroscopy (GC–MS) method that has a complete run time of only 4.5 min was developed and validated for the simultaneous determination and pharmacokinetic study of safrole and methyl eugenol in rat plasma after administration of Asarum extracts. The chromatographic separation was realized on a DB-17 column (

30 m \times 0.25 mm \times 0.25 μ m

). And detection was carried out under selected ion monitoring (SIM) mode. Plasma samples were pretreated by

n

-hexane. The pharmacokinetic parameters provided by this study will be beneficial for further developments and clinical applications of Asarum.

Link: http://dx.doi.org/10.1155/2021/6699033

Compound-Specific Chlorine Isotope Analysis of Organochlorine Pesticides by Gas Chromatography-Negative Chemical Ionization Mass Spectrometry

Compound-specific stable chlorine isotope analysis (CSIA-Cl) is an important method for identifying sources of organochlorine contaminants and helping assess their quantification of transformation processes. However, the present CSIA-Cl is challenged by either redundant conversion pretreatment or complicated mathematical correction. To overcome the mentioned problems, a novel method has been developed for the CSIA-Cl of eight organochlorine pesticides using gas chromatography-negative chemical ionization mass spectrometry (GC-NCI-qMS) in this study. The instrument parameters, acquisition mode, and required injection amounts were optimized in terms of the precision of GC-NCI-qMS. An ionization energy of 90 eV and emission current of 90 μA were selected, and the precisions for eight organochlorine pesticides were in the range of 0.37‰–2.15‰ in single ion monitoring (SIM) mode when the injected amount was 0.50 mg L−1 (viz. 0.5 ng on column). Furthermore, when standards from Supelco and O2si were calibrated using standards from AccuStandard regarded as external isotope standard, chlorine isotope composition of α-hexachlorocyclohexane (α-HCH) and 2, 2-dichloro−1, 1-bis (4-chlorophenyl) ethylene (p, p′-DDE) in Supelco and O2si was confidently differentiated. The provenance identification method was validated by three organochlorine contaminated groundwater samples and showed a prospect in identifying the source of organochlorine pesticides.

Link: http://dx.doi.org/10.1155/2021/8874679

Genotype by Environment Interaction Effects on Nutraceutical Lipid Compounds of Pigmented Rice (Oryza sativa L. ssp. indica)

To identify a rice genotype with a wider or specific adaptability, the magnitude and patterns of genotype interaction with the environment are valuable clues. However, such information on the nutraceutical lipid compound in rice is limited. In this study, concentrations of five pigmented rice genotypes with different lipid compound levels were evaluated for α-tocopherol, γ-oryzanol, phytosterols, octacosanol, and squalene contents. The research was conducted in eight environments (four locations and two years). Combined analysis of variance showed that levels of α-tocopherol, γ-oryzanol, octacosanol, and squalene were mainly affected by genotype, accounting for more than 42.57% of the total variance. This suggests that improvement of these traits may be achieved by using conventional rice breeding methods. A large proportion of the variation in campesterol and β-sitosterol contents was contributed by the environment. The effect of genotype by environment was significant for all observed traits, especially for γ-oryzanol, β-sitosterol, and squalene. This complicates identification of superior genotypes for these compounds across all environments. Stability parameters showed rice genotypes with high levels of nutraceutical lipid compounds to be sensitive to changes in environmental conditions; thus, specific adaptation is suitable. Our findings suggest that rice breeders and producers should be testing and selecting rice genotypes possessing high added-value lipid concentrations in specific growing conditions or regions.

Link: http://dx.doi.org/10.1155/2021/8880487

Comprehensive Analysis of the Components of Walnut Kernel (Juglans regia L.) in China

The contents of main components in 45 walnut kernels from 5 walnut-planting provinces in China (Yunnan, Shaanxi, Shandong, Hebei, and Sichuan) were determined using colorimetry, high-performance liquid chromatography, and gas chromatography, including flavonoids, vitamin E, trace elements, fatty acids, and amino acids. The levels of flavonoids and vitamin E were higher in walnuts from Yunnan than in walnuts from the other four provinces. The levels of zinc and iron were relatively higher in walnuts from Shandong, and the calcium content in walnuts from Yunnan was much lower. No obvious difference was found in crude fat concentration for nuts from the five provinces, but differences were observed in the crude protein content. Oleic acid was the predominant unsaturated fatty acids in all walnut species; the monounsaturated fatty acid content was the highest in walnuts from Yunnan. The data obtained here provided insight into differences in walnuts resulting from different growing environments and germplasm genetic traits.

Link: http://dx.doi.org/10.1155/2021/9302181

Performance and emission characteristics of a diesel engine employing straight vegetable oils from Vanuatu as fuels

The performance characteristics of the engine and the emission levels with Copra Oil (CPO), Virgin Coconut (cocos nucifera) Oil (VCO), Tamanu (calaphyllum inopyllum) Oil (TMO), and Nangae (canarium indicum) Oil (NGO) are presented. The oils, obtained from naturally grown trees in Vanuatu, were tested as straight vegetable oils (SVOs) in a Diesel engine and the results are compared with those of neat diesel. The oils were converted to their fatty-acid-methyl-esters (FAMEs) using gas chromatography to determine their fatty acid compositions. The brake thermal efficiency with SVOs was found to be comparable to diesel. The structure of the alkyl chain and the carbon-to-hydrogen ratio were also studied. All the oils have Palmitic acid, Capric acid, Caprylic acid, and Oleic acid as the major fatty acids. The CPO and VCO have higher amounts of Oleic acid, which acts as an additive and breaks up the interaction between the major fatty acids at higher temperatures, reducing the viscosity. Emissions of CO2 were lower while those of CO, NOx, and SO2 were higher with SVOs compared to diesel. The results indicate that the local SVOs are good and inexpensive substitute fuels for Vanuatu that can help the country meet the UN’s sustainable development goals.

Link: http://dx.doi.org/10.1177/1687814020962351

Volatile Organic Compounds to Identify Infectious (Bacteria/Viruses) Diseases of the Central Nervous System - A Pilot Study

Abstract Background Central nervous system (CNS) infectious diseases are common diseases in emergency rooms and neurology departments. CNS pathogen identification methods are time-consuming and expensive and have low sensitivity and poor specificity. Some studies have shown that bacteria and viruses can produce specific volatile organic compounds (VOCs). The aim of this study is to find potential biomarkers by VOC analysis of cerebrospinal fluid (CSF) in patients with bacterial and viral meningitis/encephalitis (ME). Methods CSF samples from 16 patients with bacterial ME and 42 patients with viral ME were collected, and solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) was used to analyze the metabolites in the CSF. Results There are two substances (Ethylene oxide and Phenol) that were found to be different between the two groups. Ethylene oxide was significantly greater in the group of bacterial ME patients than in the viral ME group of patients (P < 0.05). In addition, phenol was remarkably increased in the group of ME patients compared with the bacterial ME patients (P < 0.05). Conclusions Ethylene oxide and phenol may be potential biomarkers to distinguish bacterial ME and viral ME. VOC analysis of CSF may be used as a supporting tool for clinical diagnosis.

Link: http://dx.doi.org/10.21203/rs.2.10462/v1

Value of Amniotic Fluid Homocysteine Assay in Prenatal Diagnosis of Combined Methylmalonic Acidemia and Homocystinuria, Cobalamin C Type

Abstract Background: Combined methylmalonic acidemia and homocystinuria, cobalamin C type (cblC defect) is the most common inborn error of cobalamin metabolism. Even with rapid diagnosis and treatment, the long-term outcome remains poor. A reliable method for the prenatal diagnosis of cblC defect is needed for parental decisions regarding continuation of pregnancies of cblC defect fetuses.Results: The between-day and within-day imprecision of Hcy assay were 1.60%∼5.87% and 1.11%∼4.31%, respectively. For the 248 at-risk fetuses, there were 63 affected fetuses with cblC defect and 185 unaffected fetuses. The levels of Hcy in 63 affected fetuses were significantly higher than those in 185 unaffected fetuses, without overlap between the affected and unaffected groups. Sixteen fetuses displayed inconclusive genetic results of MMACHC variants, in which seven fetuses were determined to be affected with elevated amniotic fluid levels of Hcy, C3, C3/C2 and MMA. The remaining nine fetuses were considered unaffected based on a normal amniotic fluid metabolite profile. The diagnostic sensitivities of Hcy and other characteristic metabolites including propionylcarnitine (C3), ratio of C3 to acetylcarnitine (C2; C3/C2), methylmalonic acid (MMA), and methylcitrate acid (MCA) in amniotic fluid were 100%, 87.50%, 100%, 85.71%, and 28.57%, respectively. The respective specificities were 92.05%, 100%, 100%, 97.73%, and 99.43%. Conclusions: Hcy appears to be another sensitive characteristic metabolite biomarker for the prenatal diagnosis of cblC defect. The combination of Hcy assay with acylcarnitine and organic acid analysis is a fast, sensitive, and reliable prenatal diagnostic biochemical approach. This approach could overcome the challenge of the lack of genetic analysis for families with at-risk cblC defect fetuses.

Link: http://dx.doi.org/10.21203/rs.3.rs-108786/v1

GC–MS analysis of phytoconstituents of Acacia nilotica and its molecular docking with drug-targets of Leishmania donovani to combat leishmanaisis

Abstract Acacia nilotica is an important medicinal plant, found in Africa, Middle East, and Indian subcontinent. Every part of the plant possesses a wide array of biologically-active and therapeutically important compounds and have been used in traditional-system of medicine. We reported the antileishmanial activity of Acacia nilotica (A. nilotica) bark methanolic extract through in vitro assays and dissected the mechanism of its action through in silico studies. Bark methanolic extract exhibited anti-promastigote and anti-amastigote potency with IC50 value of 19.6 + 0.9037 µg/ml and 77.52 + 5.167 µg/ml respectively in time and dose dependent manner. It showed very low cytotoxicity having CC50 value of 432.7 + 7.71 µg/ml on human-macrophage cell line, THP-1. The major constituents identified by GC-MS analysis are 13-docosenoic acid (34.06%), lupeol(20.15 %), 9,12-octadecadienoic acid (9.92 %) and 6-octadecanoic acid (8.43 %) bind effectively with the potential drug-targets of Leishmania donovani (L. donovani ) including sterol 24-c-methytransferase (SMT), trypanothione reductase (TR), pteridine reductase (PTR1) and adenine phosphorybosyl transferase (APRT); suggest the possible mechanism of its antileishmanial action. The highest affinity with all these targets was shown by lupeol. The pharmacokinetic studies, predicted bioactivity scores and acute toxicity studies of major extract constituents support safe antileishmanial drug candidate. This study proved the antileishmanial potential of bark-methanolic extract A. nilotica and its mechanism of action through the inhibition of potential drug targets of L. donovani.

Link: http://dx.doi.org/10.21203/rs.3.rs-120320/v1

Increased Carvone Production in Escherichia Coli by Balancing Expression of Limonene Conversion Enzymes Through Targeted QconCAT Proteome Analysis

Abstract Background Natural mint flavor is produced by extraction from mint, which is not efficient enough to make it environment friendly process. (−)-Carvone is the monoterpenoid with key flavor of spearmint, and there has been an attempt to produce (−)-carvone by recombinant Escherichia coli. Although all enzymes in (−)-carvone biosynthesis have been functionally expressed in E. coli independently, the yield of (−)-carvone was low in the previous study.ResultsWe have found a by-product formation when cytochrome P450 limonene-6-hydroxylase (P450)/cytochrome P450 reductase (CPR) and carveol dehydrogenase (CDH) were expressed in single cell. These by-products were determined as dihydrocarveol and dihydrocarvone. We hypothesized that the enzymatic kinetics and the expression levels of P450 and CDH are quite different in E. coli. Therefore, two strains independently expressing P450/CPR and CDH were mixed with different mixing ratio, confirming increase in carvone production and decrease in by-product formation when CDH input was reduced. To determine the optimum balance of enzyme expressions, proteome analysis quantification concatemer (QconCAT) method to quantify P450, CPR, and CDH was developed. Using the QconCAT standard protein that was artificially created by concatenating the tryptic peptides, the ratio between P450 and CDH was calculated, and their optimum ratio to maximize (−)-carvone production was shown. Then, a single strain expressing both P450/CPR and CDH was constructed to imitate the superior expression ratio. The upgraded strain showed 15-fold improvement compared to the initial strain, showing 44 ± 6.3 mg/L of (−)-carvone production from 100 mg/L (−)-limonene as a starting substrate.ConclusionsImproved expression balance of P450 and CDH in recombinant E. coli increased the (−)-carvone production using (−)-limonene as the direct substrates by the whole-cell biocatalysis, showing approximately 150 times higher titer than previous report. Our study showed the usefulness of proteome analysis QconCAT method in the strain development for industrial biotechnology field.

Link: http://dx.doi.org/10.21203/rs.3.rs-139559/v1

Optimization of Extraction Yield and Phytochemical Characterization of Crude Methanolic Extract and Its Fractions of Mitragyna Speciosa Leaves

Abstract In this study, Response surface methodology (RSM) was applied to optimize the yield of crude methanolic extract of Mitragyna speciosa leaves using Ultrasound-assisted extraction (UAE). The crude methanolic extract and its fractions were quantified in terms of total phenolic content and total flavonoid content, along with characterized using Fourier-transform infrared and Gas chromatography–mass spectrometry. The results showed the maximum yield of 49.72% at the optimal conditions (temperature, 34 °C; time, 25 min; and volume of solvent, 166 mL). The recovery crude methanolic extract for TPC and TFC were 137.3 ± 15.7 mg GAE/g and 90.3 ± 15.3 mg RE/g, respectively.

Link: http://dx.doi.org/10.21203/rs.3.rs-52121/v1

Development of a Screening Method for Isolation of Microalgae Strains That Accumulate Lipids Under Nitrate-replete Conditions

Abstract Background: Microalgae biofuels have attracted global attention as an alternative to fossil fuels as an energy resource. Microalgae generally accumulate lipids under nitrogen-depleted conditions, but cell growth is depressed under these conditions which causes decrease in lipid productivity. To realize one-step cultivation for biofuel production, microalgae that highly accumulate lipids even under nitrogen-replete conditions are needed. This study aimed to develop a screening method for microalgae mutants with high lipid content even in the presence of a nitrogen source.Results: Mutant cells were generated by irradiating the oleaginous green microalga Chlamydomonas sp. KOR1 with carbon ion beams, cultured under nitrate-replete conditions, and then subjected to FACS-based screening for lipid-rich cells. By repeatedly performing the sequential procedures of cultivation and selection, strains KAC1710 and KAC1801, which highly accumulate lipids under nitrate-replete conditions, were successfully obtained. These mutants formed significant lipid droplets in the cells even in the presence of abundant nitrate and achieved 1.5- and 2.1-fold greater lipid content compared to KOR1, respectively.Conclusion: This study developed a novel nitrogen-conditioned screening method for microalgae mutants that accumulate lipids in the presence of a nitrogen source. This method should contribute to microalgae biofuel production via one-step cultivation under nitrogen-replete conditions.

Link: http://dx.doi.org/10.21203/rs.3.rs-86947/v1

10-Gingerol Suppresses Osteoclastogenesis in RAW264.7 Cells and Zebrafish Osteoporotic Scales

Osteoporosis is the most common aging-associated bone disease and is caused by hyperactivation of osteoclastic activity. We previously reported that the hexane extract of ginger rhizome [ginger hexane extract (GHE)] could suppress receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells. However, the anti-osteoclastic components in GHE have not yet been identified. In this study, we separated GHE into several fractions using silica gel column chromatography and evaluated their effects on osteoclastogenesis using a RAW264.7 cell osteoclast differentiation assay (in vitro) and the zebrafish scale model of osteoporosis (in vivo). We identified that the fractions containing 10-gingerol suppressed osteoclastogenesis in RAW264.7 cells detected by tartrate-resistant acid phosphatase (TRAP) staining. In zebrafish, GHE and 10-gingerol suppressed osteoclastogenesis in prednisolone-induced osteoporosis regenerated scales to promote normal regeneration. Gene expression analysis revealed that 10-gingerol suppressed osteoclast markers in RAW264.7 cells [osteoclast-associated immunoglobulin-like receptor, dendrocyte-expressed seven transmembrane protein, and matrix metallopeptidase-9 (Mmp9)] and zebrafish scales [osteoclast-specific cathepsin K (CTSK), mmp2, and mmp9]. Interestingly, nuclear factor of activated T-cells cytoplasmic 1, a master transcription regulator of osteoclast differentiation upstream of the osteoclastic activators, was downregulated in zebrafish scales but showed no alteration in RAW264.7 cells. In addition, 10-gingerol inhibited CTSK activity under cell-free conditions. This is the first study, to our knowledge, that has found that 10-gingerol in GHE could suppress osteoclastic activity in both in vitro and in vivo conditions.

Link: http://dx.doi.org/10.3389/fcell.2021.588093

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