Haylock S J, Buckley P D, Blackwell L F
J Inorg Biochem. 1983 Oct;19(2):105-17. doi: 10.1016/0162-0134(83)85016-8.
After incubation with CrCl3 X 6H2O (or 51CrCl3 X 6H2O) for 25 days, a sterile growth medium, whole yeast cells harvested after growth on a similar chromium-containing medium for the same period, and the spent growth medium remaining after removal of the yeast were each subjected to the separation procedure reported previously [S. J. Haylock. P. D. Buckley and L. F. Blackwell, J. Inorg. Biochem., in press]. The results obtained showed that most of the eleven chromium-containing fractions isolated previously were artifacts formed as a result of direct reaction between the chromium and components of the medium. An anionic complex (which was the major chromium-containing fraction isolated) was identified as a chromium-glucose complex, but one possessing no biological activity. The biologically active chromium-containing fractions (P-3 and P-4) that were only present after yeast had been grown in the medium were further purified, however, during the purification steps, the biological activity was cleanly separated from the chromium material for both P-3 and P-4. Fraction P-4 was subsequently shown to consist of approximately 90% tyramine, but pure tyramine was not active in the yeast bioassay. Although the structure of the glucose tolerance factor-active component in fraction P-3 could not be determined due to the presence of high concentrations of salt that could not be separated on gel filtration columns, the results show that the glucose tolerance factor from brewer's yeast can no longer be regarded as a chromium complex.
用CrCl₃·6H₂O（或⁵¹CrCl₃·6H₂O）孵育25天后，将无菌生长培养基、在相同含铬培养基上生长相同时间后收获的全酵母细胞以及去除酵母后剩余的用过的生长培养基分别按照先前报道的分离程序进行处理[S. J. 海洛克、P. D. 巴克利和L. F. 布莱克韦尔，《无机生物化学杂志》，即将发表]。所得结果表明，先前分离出的11个含铬组分中的大多数是铬与培养基成分直接反应形成的假象。一种阴离子络合物（这是分离出的主要含铬组分）被鉴定为铬 - 葡萄糖络合物，但不具有生物活性。仅在酵母在培养基中生长后才出现的具有生物活性的含铬组分（P - 3和P - 4）进一步纯化，然而，在纯化步骤中，P - 3和P - 4的生物活性都与铬物质清晰分离。随后表明组分P - 4约由90%的酪胺组成，但纯酪胺在酵母生物测定中无活性。尽管由于在凝胶过滤柱上无法分离的高浓度盐的存在，无法确定组分P - 3中葡萄糖耐量因子活性成分的结构，但结果表明啤酒酵母中的葡萄糖耐量因子不能再被视为铬络合物。
