Angaïts Ange, Bierla Katarzyna, Szpunar Joanna, Lobinski Ryszard
CNRS, Institute of Analytical and Physical Chemistry for the Environment and Materials (IPREM), UMR5254, Hélioparc, Universite of Pau, 64053, Pau, France.
Faculty of Chemistry, Warsaw University of Technology, ul. Noakowskiego 3, 00-664, Warsaw, Poland.
Anal Bioanal Chem. 2024 Sep;416(23):5111-5120. doi: 10.1007/s00216-024-05448-w. Epub 2024 Jul 30.
The complete characterization of selenium-enriched yeast in terms of selenium species has been the goal of extensive research for the last three decades. This contribution addresses the two outstanding questions: the mass balance of the identified and reported selenium species and the possible presence of inorganic selenium. For this purpose, four procedures have been designed combining, in diverse order, the principal steps of selenium speciation analysis in Se-rich yeast: extraction of the Se-metabolome, derivatization of cysteine and Se-cysteine (SeCys) residues, proteolysis, and definitive Se recovery using SDS extraction, followed by mineralization. The recovery of selenium in each step and its speciation were controlled by ICP MS and by reversed-phase HPLC-ICP MS, respectively. The study, carried out for the SELM-1 reference material, demonstrated the presence of about 10% of inorganic selenium and a serious risk of losses of SeCys during derivatization and proteolysis. As result of our work, we postulate the following values for SELM-1: Se-metabolome fraction (SeMF) 14.8 ± 0.7%; total selenomethionine (SeMet) 66.2 ± 2.7% (including ca. 1.5% SeMet present in the SeMF); total SeCys 12.5 ± 1.5% (including 2% of SeCys present in the Se-MF); total inorganic selenium 9.7 ± 1.7%, accounting for > 99.8% of the selenium.
在过去三十年里,从硒形态方面对富硒酵母进行全面表征一直是广泛研究的目标。本论文探讨了两个突出问题:已鉴定和报道的硒形态的质量平衡以及无机硒的可能存在情况。为此,设计了四种方法,以不同顺序组合了富硒酵母中硒形态分析的主要步骤:提取硒代谢组、半胱氨酸和硒代半胱氨酸(SeCys)残基的衍生化、蛋白质水解,以及使用SDS提取随后进行矿化的最终硒回收。每个步骤中硒的回收率及其形态分别通过电感耦合等离子体质谱(ICP MS)和反相高效液相色谱-电感耦合等离子体质谱(RP-HPLC-ICP MS)进行控制。对SELM-1参考物质进行的研究表明,存在约10%的无机硒,并且在衍生化和蛋白质水解过程中SeCys有严重损失的风险。作为我们工作的结果,我们假定SELM-1的以下值:硒代谢组分数(SeMF)14.8±0.7%;总硒代蛋氨酸(SeMet)66.2±2.7%(包括SeMF中约1.5%的SeMet);总SeCys 12.5±1.5%(包括Se-MF中2%的SeCys);总无机硒9.7±1.7%,占硒的>99.8%。