Pichia pastoris expression system

Macauley-Patrick, S., Fazenda, M.L., McNeil, B. and Harvey, L.M. (2005) Heterologous protein production using the Pichia pastoris expression system. Yeast, 22, 249-270. 

Various expression hosts were evaluated for the heterologous production of GO. We, and others (72), found that in spite of the post-translational modifications that are necessary to obtain active GO, efficient expression of GO was possible using the Pichia pastoris expression system that is commercially available from Invitrogen, Carlsbad, CA. 

With heat-dried cells of T. intermedius as PDH source and heat-dried C. boidinii as FDH source, reaction yields of 84% and ee >98% were achieved. As a major drawback of this system the production of T. intermedius could not be scaled up. To solve this problem heat-dried E. coli containing T. intermedius PDH and heat-dried C. boidinii as source of FDH were used. In this procedure, recovery was not a problem and 197 kg allysine ethylene acetal were synthesized in three batches with an average yield of 91% and ee >8%. In a third approach, recombinant Thermo-actinomyces PDH as well as heat-dried methanol-grown Pichia pastoris expressing endogenous FDH were used to produce 15 kg allysine ethylene acetal with a yield of 97% and ee >98%. This process allowed both enzymes to be produced during a... 

Although proteins can be expressed in many heterologous production systems, including bacteria such as Proteus mirabilis [1], fungi such as Pichia pastoris [2, 3] and Aspergillus awamori [4] and insect cells [5, 6], the pharmaceutical industry has narrowed down process development to a small number of platform technologies ... 

H. Lehrach, and D. Cahill, A system for dual protein expression in Pichia pastoris, Prot. Expr. Purif. 2000, 20, 372-378. 

A system for dual protein expression in Pichia pastoris and Escherichia coli. 

The system was used in three different configurations (i) the system with heat-dried cells from Th. intermedius (PheDH) and C. boidinii (FDH) yielded on average only 84 m% and could not be scaled up owing to lysis of the Th. intermedius cells (ii) a similar system with recombinant PheDH from E. coli improved the yield to 91 m% (iii) heat-dried Pichia pastoris containing endogeneous FDH and expressing recombinant PheDH from Th. intermedius yielded 98 m% with an optical purity of >98%. 

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