Fahnestock S R, Yao Z, Bedzyk L A
DuPont Experimental Station, DuPont Central Research and Development, P.O. Box 80328, Wilmington, DE 19880-0328, USA.
J Biotechnol. 2000 Aug;74(2):105-19. doi: 10.1016/s1389-0352(00)00008-8.
The remarkable properties of spider dragline silk and related protein polymers will find many applications if the materials can be produced economically. We have demonstrated the production of high molecular weight spider dragline silk analog proteins encoded by synthetic genes in several microbial systems, including Escherichia coli and Pichia pastoris. In E. coli, proteins of up to 1000 amino acids in length could be produced efficiently, but the yield and homogeneity of higher molecular weight silk proteins were found to be limited by truncated synthesis, probably as a result of ribosome termination errors. No such phenomenon was observed in the yeast P. pastoris, where higher molecular weight silk proteins could be produced without heterogeneity due to truncated synthesis. Spider dragline silk analog proteins could be secreted by P. pastoris when fused to both the signal sequence and N-terminal pro-sequence of the Saccharomyces cerevisiae alpha-mating factor gene.
如果能够经济地生产出蜘蛛拖牵丝及相关蛋白质聚合物材料，那么其卓越的性能将会有众多应用。我们已经证明，通过合成基因编码的高分子量蜘蛛拖牵丝类似蛋白能在包括大肠杆菌和巴斯德毕赤酵母在内的多种微生物系统中生产。在大肠杆菌中，可以高效生产长度达1000个氨基酸的蛋白质，但发现高分子量丝蛋白的产量和均一性受到截短合成的限制，这可能是核糖体终止错误的结果。在酵母巴斯德毕赤酵母中未观察到这种现象，在该酵母中可以生产高分子量丝蛋白且不会因截短合成而出现异质性。当与酿酒酵母α-交配因子基因的信号序列和N端前序列融合时，巴斯德毕赤酵母能够分泌蜘蛛拖牵丝类似蛋白。
