Highly efficient secretion of heterologous proteins from Saccharomyces cerevisiae using inulinase signal peptides

✍ Scribed by Bong Hyun Chung; Soo Wan Nam; Byung Moon Kim; Young Hoon Park

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 653 KB
• Volume: 49
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/(sici)1097-0290(19960220)49:4<473::aid-bit15>3.0.co;2-b

✦ Synopsis

The INU genes o f Kluyveromyces marxianus encode inulinases which are readily secreted from Saccharomyces cerevisiaeinto the culture medium. To evaluate the utility of the INU signal peptides for the secretion of heterologous proteins from S. cerevisiae, a variety of expression and secretion vectors were constructed with GAL70 promoter and GAL7 terminator. The coding sequence for human lipocortin-I (LCI) was inserted in-frame with the INU signal sequences, and then the secretion efficiency and localization of LC1 were investigated in more detail and compared with those when being expressed by the vector with the MFa 1 leader peptide. The vector systems with INU signal peptides secreted ca. 95% of the total LC1 expressed into the extracellular medium, while the MFa 7 leader peptide-containing vector resulted in very low secretion efficiency below 10%. In addition, recombinant human interleukin-2 (IL-2) was expressed and secreted with the vector systems with INU signal peptide, and a majority fraction of the human IL-2 expressed was found to be secreted into the extracellular medium as observed in LC1 expression. 0 1995 John Wiley &Sons, Inc.