Buffered Non-Fermenter System for Lab-Scale Production of Secreted Recombinant His-Tagged Proteins in Saccharomyces cerevisiae | Zendy

Chatri Ngamkitidechakul | Zendy; Sally S. Twining | Zendy

Open Access

Buffered Non-Fermenter System for Lab-Scale Production of Secreted Recombinant His-Tagged Proteins in Saccharomyces cerevisiae

Author(s) -

Chatri Ngamkitidechakul,

Sally S. Twining

Publication year - 2002

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/02336pt02

Subject(s) - industrial fermentation , saccharomyces cerevisiae , recombinant dna , biology , biochemistry , microbiology and biotechnology , chemistry , yeast , fermentation , gene

Expression of recombinant proteins using a secretion system can minimize co-purification of contaminating host proteins. Production of His-tagged recombinant proteins in the yeast alpha-factor secretion system has previously required a fermenter system to control the growth conditions such as pH of the yeast culture. We describe an inexpensive non-fermenter system for the production of secreted recombinant His-tagged proteins in Saccharomyces cerevisiae that uses a buffered low peptone YP glycerol medium, which does not interfere with immobilized metal affinity chromatography. Maspin, a tumor suppressor serpin, was expressed as a secreted N-terminal His/FLAG-tagged protein. Purification of the soluble active recombinant protein only requires centrifugation, concentration by ultrafiltration, and Ni2+ affinity chromatography. Purified protein yields of this system are 3-5 mg/L culture medium.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research