Cutinase-like biodegradable plastic-degrading enzymes from phylloplane yeasts have cutinase activity

2021 ◽  
Author(s):  
Hirokazu Ueda ◽  
Jun Tabata ◽  
Yasuyo Seshime ◽  
Kazuo Masaki ◽  
Yuka Sameshima-Yamashita ◽  
...  
Keyword(s):  
Biodegradable Plastic ◽  
Gas Chromatography Mass Spectrometry ◽  
Cryptococcus Laurentii ◽  
Amino Acid Sequence Homology ◽  
Degrading Enzymes ◽  
Reaction Solution ◽  
Catalytic Sites ◽  
Cryptococcus Species ◽  
Phylloplane Yeasts ◽  
Layer Chromatography

Abstract Phylloplane yeast genera Pseudozyma and Cryptococcus secrete biodegradable plastic (BP)-degrading enzymes, termed cutinase-like enzymes (CLEs). Although, CLEs contain highly conserved catalytic sites, the whole protein exhibits ≤ 30% amino acid sequence homology with cutinase. In this study, we analyzed whether CLEs exhibit cutinase activity. Seventeen Cryptococcus magnus strains, which degrade BP at 15 °C, were isolated from leaves, and identified the DNA sequence of the CLE in one of the strains. Cutin was prepared from tomato leaves and treated with CLEs from three Cryptococcus species (C. magnus, Cryptococcus flavus, and Cryptococcus laurentii) and Pseudozyma antarctia (PaE). A typical cutin monomer, 10,16-dihydroxyhexadecanoic acid, was detected in extracts of the reaction solution via gas chromatography-mass spectrometry, showing that cutin was indeed degraded by CLEs. In addition to the aforementioned monomer, separation analysis via thin-layer chromatography detected high-molecular-weight products resulting from the breakdown of cutin by PaE, indicating that PaE acts as an endo-type enzyme.

scholarly journals Cryptococcus Species Other Than Cryptococcus neoformans and Cryptococcus gattii: Are They Clinically Significant?

2020 ◽  
Vol 7 (12) ◽  
Author(s):  
Edison J Cano ◽  
Zachary A Yetmar ◽  
Raymund R Razonable
Keyword(s):  
Cryptococcus Neoformans ◽  
Opportunistic Infections ◽  
Descriptive Analysis ◽  
Laboratory Data ◽  
Invasive Disease ◽  
Cryptococcus Gattii ◽  
Cryptococcus Laurentii ◽  
Cryptococcus Species ◽  
Clinically Significant ◽  
Cryptococcus Spp

Abstract Background Cryptococcus spp is a major cause of opportunistic infections in immunocompromised patients, primarily due to Cryptococcus neoformans and Cryptococcus gattii. There are occasional reports of other Cryptococcus species causing invasive human disease. However, their epidemiology and clinical significance are not fully defined. We sought to describe cases with cultures positive for Cryptococcus species other than C neoformans and C gattii. Methods A retrospective descriptive analysis of clinical and laboratory data of patients with cultures growing Cryptococcus species other than C neoformans and C gattii from November 2011 to February 2019 was performed. Three Mayo Clinic sites in Arizona, Florida, and Minnesota were included. Results From 176 cases with a culture growing Cryptococcus spp, 54 patients (30%) had a culture for Cryptococcus other than C neoformans and C gattii in the study time frame. The most common species were Cryptococcus magnus, Cryptococcus laurentii, and Cryptococcus ater. The organisms were isolated and identified in culture of bronchoalveolar lavage (11), skin (11), urine (7), oral (4), sinus (3), intraoperative soft tissue (3), sputum (2), synovial fluid (2), cerebrospinal fluid (2), and intravenous catheter (2), among others (7). Only 8 (15%) cases were considered to be potentially pathogenic, with 1 case of invasive disease. Antifungal treatment was fluconazole, itraconazole, and griseofulvin, for a mean systemic antifungal duration of 42 days. Conclusions This large series of patients with Cryptococcus spp other than C neoformans and C gattii suggests that these species rarely cause clinically significant infection in humans. Only 1 case of invasive disease was found.

Analytical and technical aspects of testing for drug abuse: confirmatory procedures.

1988 ◽  
Vol 34 (3) ◽  
pp. 471-473 ◽  
Author(s):  
M A Peat
Keyword(s):  
Drug Testing ◽  
Analytical Procedure ◽  
Gas Chromatography Mass Spectrometry ◽  
Technical Aspects ◽  
Governmental Agencies ◽  
Accuracy And Precision ◽  
Urine Drug Testing ◽  
Urine Drug ◽  
Chemical Technique ◽  
Layer Chromatography

Abstract Many laboratories are now performing urine drug testing for employers, governmental agencies, and other institutions. It is now recognized that presumptive positive screening results have to be confirmed by an analytical procedure based on a different chemical technique with greater than or equal sensitivity to the screening test. Thin-layer chromatography has been widely used for this; however, it is relatively insensitive for certain drugs, and it cannot satisfy the accuracy and precision requirements needed to determine threshold concentrations reliably. Gas chromatography-mass spectrometry is able to satisfy these threshold requirements and has become the method of choice for confirming initial immunoassay results.

Thin-layer chromatography and gas chromatography-mass spectrometry examination of shoe materials from patients with shoe dermatitis

2015 ◽  
Vol 72 (4) ◽  
pp. 248-252 ◽  
Author(s):  
Sri A. Febriana ◽  
Erik Zimerson ◽  
Cecilia Svedman ◽  
Winarto Haryadi ◽  
Pieter-Jan Coenraads ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Gas Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Layer Chromatography

Detection of the toxic substance dibutyl phthalate in Antarctic krill

2017 ◽  
Vol 29 (6) ◽  
pp. 511-516 ◽  
Author(s):  
Xiangning Han ◽  
Daicheng Liu
Keyword(s):  
High Performance ◽  
Dibutyl Phthalate ◽  
Antarctic Krill ◽  
Toxic Substance ◽  
Dioctyl Phthalate ◽  
Gas Chromatography Mass Spectrometry ◽  
Coastal Marine ◽  
Freeze Dried ◽  
Coastal Marine Ecosystems ◽  
Layer Chromatography

AbstractHigh-performance thin layer chromatography was performed to investigate the potential presence of four phthalic acid esters, dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP) and dioctyl phthalate (DEHP), in Antarctic krill. The results revealed that in freeze-dried Antarctic krill levels of DBP (0.1043±0.0005 mg g-1 (104.3±0.05 mg kg-1)) were high. The structure of DBP in Antarctic krill was determined by gas chromatography-mass spectrometry. Its existence is of concern based on demonstrated harmful effects to animals and plants as Antarctic krill is a key part of the food chain in Antarctic coastal marine ecosystems. The adverse effects of DBP on Antarctic krill and the source of DBP should be explored in further research.

Extraction and Characterization of Gibberellins from Hordeum vulgare L. Seedlings

1974 ◽  
Vol 1 (2) ◽  
pp. 183 ◽  
Author(s):  
KF Faull ◽  
BG Coombe ◽  
LG Paleg
Keyword(s):  
Mass Spectrometry ◽  
Dry Weight ◽  
Paper Electrophoresis ◽  
The Other ◽  
Gas Chromatography Mass Spectrometry ◽  
Gas Liquid Chromatography ◽  
Fresh Weight ◽  
Hordeum Vulgare L ◽  
Layer Chromatography

Two gibberellins, one GA1-like, the other GA3-like, were identified in the extracts of roots and tops of 8-,11- and 15-day-old barley seedlings by paper chromatography, paper electrophoresis, thin-layer chromatography, gas-liquid chromatography and bioassay procedures, followed by combined gas chromatography-mass spectrometry. The amounts of gibberellins in the seedlings ranged from 7 to 11 ng per plant. The concentrations of gibberellins in the seedlings were 32-320 ng/g dry weight and 5-28 ng/g fresh weight; concentrations in the roots were higher than those in the shoots.

Efficient extraction of basic, neutral, and weakly acidic drugs from plasma for analysis by gas chromatography-mass spectrometry

1991 ◽  
Vol 37 (11) ◽  
pp. 1975-1978 ◽  
Author(s):  
K E Brooks ◽  
N B Smith
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Acetic Anhydride ◽  
Mobile Phase ◽  
Plasma Sample ◽  
Gas Chromatography Mass Spectrometry ◽  
Acidic Drugs ◽  
Chromatography Mass Spectrometry ◽  
Efficient Extraction ◽  
Layer Chromatography

Abstract We describe a method for efficiently extracting basic, neutral, and weakly acidic drugs from plasma for toxicological analysis by gas chromatography-mass spectrometry (GC/MS). The 2-mL plasma sample is diluted with an equal volume of saturated NaCl containing triethylamine, 10 mmol/L, and then extracted twice with 4 mL of an equivolume solution of dichloromethane/acetone. The organic (top) phases are combined, then mixed with 1 mL of water, 200 mg of NaHCO3, and 100 microliters of acetic anhydride. This mixture is then heated at 75 degrees C until the solvents have boiled off and aqueous acetylation is complete (less than 30 min). After addition of 1 mL of water and 2 g of NaCl, the sample is extracted twice with 2 mL of dichloromethane/acetone (2/1 by vol). The combined extracts are dried and then subjected to thin-layer chromatography on a blank Toxi-Lab Toxi-A chromatogram with 1-chlorobutane as the developing solvent (about 6 min). After the lipids have migrated with the mobile phase, the drugs are eluted from the origin with acetone/triethylamine (29/1 by vol), evaporated, and reconstituted in injection solvent. With this procedure drugs are recovered relatively quickly (less than 2 h) and the GC/MS total ion chromatograms are very clean. Studies with 13 basic, neutral, and weakly acidic drugs showed that all except theophylline were extracted with recoveries of at least 75%.

Gas Chromatographic Determination of Low Levels of Di-(2-ethylh exyl)phthalate in Soy Oil

1973 ◽  
Vol 56 (1) ◽  
pp. 181-183
Author(s):  
D T Williams
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Chromatographic Determination ◽  
Phthalate Ester ◽  
Gas Chromatography Mass Spectrometry ◽  
Soy Oil ◽  
Chromatography Mass Spectrometry ◽  
Cleanup Procedure ◽  
Layer Chromatography ◽  
Ethylhexyl Phthalate

Abstract Di-(2-ethylhexyl)phthalate can be determined by gas chromatography at a level of 0.3 ppm in soy oil. A cleanup procedure involving epoxidation of soy oil residues with m-chloroperbenzoic acid allows confirmation of the phthalate ester by thin layer chromatography-mass spectrometry or gas chromatography-mass spectrometry.

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