Köhler P, Belitz H D, Wieser H
Deutsche Forschungsanstalt für Lebensmittelchemie, München, Federal Republic of Germany.
Z Lebensm Unters Forsch. 1991 Mar;192(3):234-9. doi: 10.1007/BF01202885.
Gluten from the wheat variety Rektor was extracted with 70% aqueous ethanol. The residual protein (glutenin) was hydrolysed with trypsin. The partial hydrolysate was separated into seven fractions by gel permeation chromatography on Sephadex G 25. Four cystine-containing peptides were isolated from fraction 5 by reversed-phase high-performance liquid chromatography on ODS-Hypersil. The cystine peptides were detected by differential chromatography of the non-reduced and the reduced samples. The primary structure of the peptides was solved by the Edman degradation reaction and by partial hydrolysis with thermolysin. Three peptides derive from the alpha 2- and beta-purothionins. The structure of the fourth peptide was determined as (Formula; see text) This sequence corresponds to positions 44-48 of known sequences of the high molecular weight (HMW) subunits 9, 10, and 12. Since Rektor contains the HMW subunits 9 and 10, it could be concluded that two HMW subunits 9 or 10, or one subunit 9 and one subunit 10 were linked parallel via two disulphide bridges.
从小麦品种雷克托（Rektor）中提取的麸质用70%的乙醇水溶液进行萃取。残留蛋白质（谷蛋白）用胰蛋白酶进行水解。部分水解产物通过在葡聚糖凝胶G 25上进行凝胶渗透色谱法分离成七个组分。通过在ODS-高效硅胶上进行反相高效液相色谱法从组分5中分离出四种含胱氨酸的肽段。通过对未还原和还原样品进行差异色谱法检测胱氨酸肽段。通过埃德曼降解反应和用嗜热菌蛋白酶进行部分水解确定肽段的一级结构。三种肽段来源于α2-和β-硫堇蛋白。确定第四种肽段的结构为（分子式；见原文）。该序列对应于高分子量（HMW）亚基9、10和12已知序列的第44 - 48位。由于雷克托含有HMW亚基9和10，可以得出结论，两个HMW亚基9或10，或者一个亚基9和一个亚基10通过两个二硫键平行连接。
