Description
The study aimed to obtain conditions optimal for the activity of Candida albicans cystathionine-γ-synthase (CaStr2p) determination. The selection of appropriate reaction conditions included the identification of the reaction buffer and its pH, substrate, and enzyme concentrations. The activity of Str2p was measured by the detection of decrease of the substrate L-cysteine (L-Cys) via LC/MS system with pre-column derivatization. Mass spectra were recorded using an Agilent 6470A triple quadrupole (6470 Triple Quad LC/MS, Agilent Technologies, Waldbronn, Germany) LC/MS system (1260 Infinity II, Agilent Technologies,Waldbronn, Germany) with electrospray ionization source (ESI). L-Cys reacts with 5,5’-dithiobis(2-nitrobenzoic acid) (DTNB) to produce a L-Cys-TNB complex, that can be also detected by the diode array detector (HPLC-DAD). Optimal conditions for L-Cys-TNB complex detection included identification of mobile phase, elution profile, injection volume, and most suitable wavelengths for the detection have been identified.
This dataset contains the description of the methodology, and the set of selected chromatographs obtained for control as well as enzymatic reaction samples.
The studies were financially supported by the project OPUS 20 financed by the National Science Centre (No. UMO-2020/39/B/NZ7/01519).
Dataset file
hexmd5(md5(part1)+md5(part2)+...)-{parts_count}
where a single part of the file is 512 MB in size.Example script for calculation:
https://github.com/antespi/s3md5
File details
- License:
-
open in new tabCC BYAttribution
- Raw data:
- Data contained in dataset was not processed.
Details
- Year of publication:
- 2024
- Verification date:
- 2024-06-14
- Dataset language:
- English
- Fields of science:
-
- chemical sciences (Natural sciences)
- DOI:
- DOI ID 10.34808/st1j-4j69 open in new tab
- Funding:
- Verified by:
- Gdańsk University of Technology
Keywords
References
- dataset Activity assay of O-Acetyl-L-homoserine sulfhydrylase (CaMet15p).
- dataset Identification and cloning of C. albicans SC5314 genes encoding L-methionine biosynthetic pathway enzymes.
- dataset Overproduction of CaMet15p native and His-tag versions.
- dataset Overproduction of homoserine O-acetyltransferase (CaMet2p) native and His-tag versions.
- dataset L-Homoserine O-acetyltransferase (CaMet2p) activity and inhibition
- dataset Overproduction of CaStr2p native and His-tag versions.
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