scholarly journals Biodegradation of Chloroxylenol by Cunninghamella elegans IM 1785/21GP and Trametes versicolor IM 373: Insight into Ecotoxicity and Metabolic Pathways

2021 ◽  
Vol 22 (9) ◽  
pp. 4360
Author(s):  
Marta Nowak ◽  
Katarzyna Zawadzka ◽  
Janusz Szemraj ◽  
Aleksandra Góralczyk-Bińkowska ◽  
Katarzyna Lisowska
Keyword(s):  
Trametes Versicolor ◽  
Significant Rise ◽  
Pcr Analysis ◽  
Cunninghamella Elegans ◽  
Oxidation Reactions ◽  
Cytochrome P450 Enzymes ◽  
C Elegans ◽  
Real Time Quantitative Pcr ◽  
Genes Expression ◽  
By Products

Chloroxylenol (PCMX) is applied as a preservative and disinfectant in personal care products, currently recommended for use to inactivate the SARS-CoV-2 virus. Its intensive application leads to the release of PCMX into the environment, which can have a harmful impact on aquatic and soil biotas. The aim of this study was to assess the mechanism of chloroxylenol biodegradation by the fungal strains Cunninghamella elegans IM 1785/21GP and Trametes versicolor IM 373, and investigate the ecotoxicity of emerging by-products. The residues of PCMX and formed metabolites were analysed using GC-MS. The elimination of PCMX in the cultures of tested microorganisms was above 70%. Five fungal by-products were detected for the first time. Identified intermediates were performed by dechlorination, hydroxylation, and oxidation reactions catalysed by cytochrome P450 enzymes and laccase. A real-time quantitative PCR analysis confirmed an increase in CYP450 genes expression in C. elegans cells. In the case of T. versicolor, spectrophotometric measurement of the oxidation of 2,20-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) showed a significant rise in laccase activity during PCMX elimination. Furthermore, with the use of bioindicators from different ecosystems (Daphtoxkit F and Phytotoxkit), it was revealed that the biodegradation process of PCMX had a detoxifying nature.

Study on the expression of specific chimeric-orfs of C type cytoplasmic male sterile in maize

2013 ◽  
Vol 61 (1-4) ◽  
pp. 25-36 ◽  
Author(s):  
Jiyue Wang ◽  
Lihua Zeng ◽  
Shengqing Wang ◽  
Jing Wang ◽  
Yanli Lu ◽  
...  
Keyword(s):  
Base Substitution ◽  
Pcr Analysis ◽  
Male Sterile ◽  
Maintainer Line ◽  
Cytoplasmic Male Sterile ◽  
Real Time Quantitative Pcr ◽  
Substitution Mutations ◽  
Membrane Spanning ◽  
Insertion Mutations ◽  
Online Software

In the present study, a mutated chimericorf364, namedorf366-c, was identified in a maize C type cytoplasmic male sterile line (CMS-C) by PCR and RT-PCR. Four base pair adjacent base substitution mutations (CAAA to TTTT) and eight bp insertion mutations were found inorf366-ccompared withorf364. ORF366-C was predicted to contain one membrane-spanning domain using TMHMM online software. Real-time quantitative PCR analysis showed thatorf366-cwas upregulated in the CMS-C line in comparison to its maintainer line at the uninucleate stage. The protokaryotic protein expression oforf366-cinEscherichia colishowed that ORF366-C could be a cytotoxic protein. All the results indicated thatorf366-cmay be associated with maize CMS-C.

scholarly journals 2C-B-Fly-NBOMe Metabolites in Rat Urine, Human Liver Microsomes and C. elegans: Confirmation with Synthesized Analytical Standards

Metabolites ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 775
Author(s):  
Jitka Nykodemová ◽  
Anna Šuláková ◽  
Petr Palivec ◽  
Hedvika Češková ◽  
Silvie Rimpelová ◽  
...  
Keyword(s):  
Phase I ◽  
Human Liver ◽  
Liver Microsomes ◽  
Human Liver Microsomes ◽  
Diagnostic Tools ◽  
Cunninghamella Elegans ◽  
Rat Urine ◽  
C Elegans ◽  
Fragmentation Patterns

Compounds from the N-benzylphenethylamine (NBPEA) class of novel psychoactive substances are being increasingly utilized in neurobiological and clinical research, as diagnostic tools, or for recreational purposes. To understand the pharmacology, safety, or potential toxicity of these substances, elucidating their metabolic fate is therefore of the utmost interest. Several studies on NBPEA metabolism have emerged, but scarce information about substances with a tetrahydrobenzodifuran (“Fly”) moiety is available. Here, we investigated the metabolism of 2-(8-bromo-2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b’]difuran-4-yl)-N-(2-methoxybenzyl)ethan-1-amine (2C-B-Fly-NBOMe) in three different systems: isolated human liver microsomes, Cunninghamella elegans mycelium, and in rats in vivo. Phase I and II metabolites of 2C-B-Fly-NBOMe were first detected in an untargeted screening and identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Several hypothesized metabolites were then synthesized as reference standards; knowledge of their fragmentation patterns was utilized for confirmation or tentative identification of isomers. Altogether, thirty-five phase I and nine phase II 2C-B-Fly-NBOMe metabolites were detected. Major detected metabolic pathways were mono- and poly-hydroxylation, O-demethylation, oxidative debromination, and to a lesser extent also N-demethoxybenzylation, followed by glucuronidation and/or N-acetylation. Differences were observed for the three used media. The highest number of metabolites and at highest concentration were found in human liver microsomes. In vivo metabolites detected from rat urine included two poly-hydroxylated metabolites found only in this media. Mycelium matrix contained several dehydrogenated, N-oxygenated, and dibrominated metabolites.

Selection of Stable Reference Genes for Real-Time Quantitative PCR Analysis in Edwardsiella tarda

2017 ◽  
Vol 27 (1) ◽  
pp. 112-121 ◽  
Author(s):  
Zhongyang Sun ◽  
Jia Deng ◽  
Haizhen Wu ◽  
Qiyao Wang ◽  
Yuanxing Zhang
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Reference Genes ◽  
Edwardsiella Tarda ◽  
Pcr Analysis ◽  
Real Time Quantitative Pcr ◽  
Selection Of

scholarly journals Piceatannol, a Natural Stilbene, Extends the Lifespan of Caenorhabditis elegans Under Fasting Through Inhibition of Lipolysis

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 38-38
Author(s):  
Jang Miran ◽  
Zhang Yuan ◽  
Bai Juan ◽  
Jun-Bae An ◽  
Park Yeonhwa ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Caenorhabditis Elegans ◽  
Negative Energy ◽  
Hormone Sensitive Lipase ◽  
Pcr Analysis ◽  
Lifespan Extension ◽  
Mrna Levels ◽  
Free Glycerol ◽  
C Elegans ◽  
Glycerol Level

Abstract Objectives Lipolysis is the catabolic process that hydrolyzes triglyceride (TG) to free fatty acids (FFAs) and glycerol under negative energy balance such as fasting. In adipocytes, adipose TG lipase (ATGL), hormone-sensitive lipase (HSL), and monoglyceride lipase play key roles in a series of TG hydrolysis reactions in mammals. However, overly activated adipose lipolysis is believed to contribute to link between obesity and systemic inflammation and oxidative stress. We previously demonstrated that piceatannol (PIC), a natural resveratrol analogue, inhibits adipogenesis in cultured adipocytes and lipogenesis in Caenorhabditis elegans. Furthermore, we showed that PIC extends the lifespan of C. elegans via the insulin/IGF-1 signaling. However, the effects of PIC on lipid metabolism during fasting state is unknown. Methods We conducted Oil-Red-O assay, Enzyme assay (TG and Free glycerol contents), PCR analysis and lifespan assay. Results In this study, we demonstrated that PIC-treated C. elegans exhibited suppressed lipolysis under fasting as judged by increased lipid accumulation and TG levels with decreased free glycerol level. Consistent with these findings, PIC treatment resulted in decreased mRNA levels of genes involved lipolysis such as atgl-1, hosl-1 and aak-2 in fasted C. elegans. Also, PIC treatment augmented fasting-induced lifespan of C. elegans by an increased daf-16 gene expression. However, such effect was abolished when atgl-1, aak-2, and daf-16 mutants were treated with PIC. In addition, we also found that autophagy is required for PIC-induced lifespan in C. elegans during fasting since autophagy inhibitor treatments and autophagy gene deficient worms resulted in blunting the lifespan extension effect of PIC. Conclusions Collectively, our results indicate that PIC contributes to lifespan extension in C. elegans during fasting possibly through regulating lipolysis- and/or autophagy-dependent lipid metabolism. Funding Sources 1. The National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (2019R1A2C1086146) and (2019R1A6A3A03033878) 2. The Rural Development Administration of the Republic of Korea.

scholarly journals Cloning of Acyl-ACP Thioesterase FatA fromArachis hypogaeaL. and Its Expression inEscherichia coli

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Gao Chen ◽  
Zhen-ying Peng ◽  
Lei Shan ◽  
Ning Xuan ◽  
Gui-ying Tang ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Palmitoleic Acid ◽  
Membrane Lipid ◽  
Pcr Analysis ◽  
Fatty Acid Profiles ◽  
Reading Frame ◽  
E Coli ◽  
Real Time Quantitative Pcr ◽  
Immature Seeds ◽  
Expression Inescherichia Coli

In this study, a full-length cDNA of the acyl-ACP thioesterase,AhFatA, was cloned from developing seeds ofArachis hypogaeaL. by 3′-RACE. Sequence analysis showed that the open reading frame encodes a peptide of 372 amino acids and has 50–70% identity with FatA from other plants. Real-time quantitative PCR analysis revealed thatAhFatA was expressed in all tissues ofA. hypogaeaL., but most strongly in the immature seeds harvested at 60 days after pegging. Heterologous expression ofAhFatA inEscherichia coliaffected bacterial growth and changed the fatty acid profiles of the membrane lipid, resulting in directed accumulation towards palmitoleic acid and oleic acid. These results indicate that AhFatA is at least partially responsible for determining the high palmitoleic acid and oleic acid composition ofE. coli.

Fate of Residual Lignin during Delignification of Kraft Pulp by Trametes versicolor

1998 ◽  
Vol 64 (6) ◽  
pp. 2117-2125 ◽  
Author(s):  
Ian D. Reid
Keyword(s):  
Trametes Versicolor ◽  
Kraft Pulp ◽  
Lignin Degradation ◽  
Water Soluble ◽  
Kraft Pulps ◽  
Residual Lignin ◽  
Spruce Wood ◽  
Single Ring ◽  
Enzyme Bleaching ◽  
By Products

ABSTRACT The fungus Trametes versicolor can delignify and brighten kraft pulps. To better understand the mechanism of this biological bleaching and the by-products formed, I traced the transformation of pulp lignin during treatment with the fungus. Hardwood and softwood kraft pulps containing 14C-labelled residual lignin were prepared by laboratory pulping of lignin-labelled aspen and spruce wood and then incubated with T. versicolor. After initially polymerizing the lignin, the fungus depolymerized it to alkali-extractable forms and then to soluble forms. Most of the labelled carbon accumulated in the water-soluble pool. The extractable and soluble products were oligomeric; single-ring aromatic products were not detected. The mineralization of the lignin carbon to CO2 varied between experiments, up to 22% in the most vigorous cultures. The activities of the known enzymes laccase and manganese peroxidase did not account for all of the lignin degradation that took place in the T. versicolor cultures. This fungus may produce additional enzymes that could be useful in enzyme bleaching systems.

scholarly journals Real-Time Quantitative PCR Analysis of the Expression Pattern of the Hypoglycemic Polypeptide-P Gene in Momordica charantia

Genes ◽  
2019 ◽  
Vol 10 (12) ◽  
pp. 1044
Author(s):  
Yi-Shuai Wang ◽  
Xiang-Qing Zeng ◽  
Xu-Zhong Yang ◽  
Wei Liu ◽  
Peng-Fei Li ◽  
...  
Keyword(s):  
Real Time ◽  
Expression Pattern ◽  
Quantitative Polymerase Chain Reaction ◽  
Gene Expression Pattern ◽  
Momordica Charantia ◽  
Expression Level ◽  
Pcr Analysis ◽  
Real Time Quantitative Pcr ◽  
P Gene ◽  
Qpcr Method

This study was designed to establish a real-time quantitative polymerase chain reaction (qPCR) method to rapidly and reliably analyze the hypoglycemic polypeptide-P gene expression pattern in Momordica charantia (MC) and to examine its expression changes in different MC accessions, harvesting seasons, and tissue types. The qPCR results were further verified by using Western blotting (WB). A total of 10 MCs with different accessions were collected and tested in this study. Among the tested accessions, RU5H showed the highest expression level of the polypeptide-P gene. The expression level of the polypeptide-P gene was not only season-related (with the highest in early July) but also tissue-related (with the highest in the seed tissue). In addition, the expression characteristic of the polypeptide-P gene was maturity-related, with the highest expression level in the tender MC. The WB results show that the transcription level of this gene shows an almost similar trend to the corresponding protein expression level. In conclusion, the established qPCR method can rapidly and effectively detect the expression levels of the polypeptide-P gene in MCs with different accessions; furthermore, various factors, including the accessions, harvesting seasons, and tissue types can affect the expression level.

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