Cazin J, Suter M, Butler J E
Department of Microbiology, University of Iowa, Iowa City 52242.
J Immunol Methods. 1988 Oct 4;113(1):75-81. doi: 10.1016/0022-1759(88)90383-3.
A simple, inexpensive procedure for producing streptavidin has been described. The biotin-binding protein was produced by growing Streptomyces avidinii in a synthetic liquid culture medium containing L-asparagine as the sole nitrogen source. With this procedure, extraneous proteinaceous substances inherently present in culture media prepared with yeast extract or with peptones were not present to interfere with isolation and purification of streptavidin. When harvested after 7-8 days of incubation, the culture fluid was relatively free of contaminating cell breakdown products. Maximal production of streptavidin (100-120 mg/l) was obtained in 8-10 day cultures. For some applications, the culture fluid can be used directly as a source of streptavidin. Under the same conditions used to grow S. avidinii, 11 other actinomycete strains and 134 eumycetes were found to lack the capacity to produce detectable amounts of an extracellular biotin-binding protein.
已描述了一种生产链霉亲和素的简单、低成本方法。通过在以L-天冬酰胺作为唯一氮源的合成液体培养基中培养阿维链霉菌来生产生物素结合蛋白。采用此方法,用酵母提取物或蛋白胨制备的培养基中固有存在的外来蛋白质物质不会干扰链霉亲和素的分离和纯化。培养7-8天后收获时,培养液相对不含污染的细胞裂解产物。在8-10天的培养物中获得链霉亲和素的最大产量(100-120毫克/升)。对于某些应用,培养液可直接用作链霉亲和素的来源。在用于培养阿维链霉菌的相同条件下,发现其他11种放线菌菌株和134种真菌缺乏产生可检测量细胞外生物素结合蛋白的能力。