Quenching Methods for the Analysis of Intracellular Metabolites

Author(s):  
Judith Wahrheit ◽  
Elmar Heinzle
2006 ◽  
Vol 61 (5-6) ◽  
pp. 347-350 ◽  
Author(s):  
Rilka M. Taskova ◽  
Holger Zorn ◽  
Ulrich Krings ◽  
Henning Bouws ◽  
Ralf G. Berger

Different techniques were compared for their effectiveness in the disruption of the rigid cell walls of Basidiomycetes. Grinding under liquid nitrogen, stirred glass bead milling and enzymatic cell lysis were applied to the mycelia of Pleurotus sapidus and Lepista irina grown submerged. Each of the disruption procedures was evaluated by testing the quantity and quality of released intracellular metabolites: DNA, RNA, enzymes, and secondary metabolites. The most suitable method for nucleic acid isolation was grinding under liquid nitrogen, while bead mill homogenization was the superior technique for isolation of active enzymes. A new effective method is proposed for isolation of secondary metabolites with the aid of bead milling of fungal mycelia.


2022 ◽  
Vol 11 (2) ◽  
pp. 332-340
Author(s):  
Pin Chen ◽  
Xiaoqian Chen ◽  
Wei Yu ◽  
Bo Zhou ◽  
Lihua Liu ◽  
...  

2012 ◽  
pp. 195-219
Author(s):  
Vineetha Zacharia ◽  
Daniel Lafontaine ◽  
Wade C. Winkler

2016 ◽  
Vol 45 (22) ◽  
pp. 6311-6326 ◽  
Author(s):  
Nathaniel W. Brown ◽  
Alan M. Marmelstein ◽  
Dorothea Fiedler

New chemical and analytical tools have been developed to study the diverse functions of the inositol pyrophosphates, a unique group of densely phosphorylated intracellular metabolites found in a wide variety of eukaryotic organisms.


Author(s):  
Jingyi Zhu ◽  
Yeyin Yang ◽  
Shunshan Duan ◽  
Dong Sun

Antialgal compounds from plants have been identified as promising candidates for controlling harmful algal blooms (HABs). In our previous study, luteolin-7-O-glucuronide was used as a promising algistatic agent to control Phaeocystis globosa (P. globose) blooms; however, its antialgal mechanism on P. globosa have not yet been elaborated in detail. In this study, a liquid chromatography linked to tandem mass spectrometry (LC-MS/MS)-based untargeted metabolomic approach was used to investigate changes in intracellular and extracellular metabolites of P. globosa after exposure to luteolin-7-O-glucuronide. Significant differences in intracellular metabolites profiles were observed between treated and untreated groups; nevertheless, metabolic statuses for extracellular metabolites were similar among these two groups. For intracellular metabolites, 20 identified metabolites showed significant difference. The contents of luteolin, gallic acid, betaine and three fatty acids were increased, while the contents of α-Ketoglutarate and acetyl-CoA involved in tricarboxylic acid cycle, glutamate, and 11 organic acids were decreased. Changes in those metabolites may be induced by the antialgal compound in response to stress. The results revealed that luteolin played a vital role in the antialgal mechanism of luteolin-7-O-glucuronide on P. globosa, because luteolin increased the most in the treatment groups and had strong antialgal activity on P. globosa. α-Ketoglutarate and acetyl-CoA were the most inhibited metabolites, indicating that the antialgal compound inhibited the growth through disturbed the tricarboxylic acid (TCA) cycle of algal cells. To summarize, our data provides insights into the antialgal mechanism of luteolin-7-O-glucuronide on P. globosa, which can be used to further control P. globosa blooms.


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