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A new dimension -
Pichia pastoris - 20 g/L - methanol-free


VTU´s 2nd generation PAOX1 library
In a recent case study VTU´s broad and versatile Pichia pastoris protein production platform, impressing with record productivities, hit the 20 g/L mark in methanol-free protein production.
VTU´s unique methanol-free Pichia system -
strong expression and further advantages such as

 
  • abolishing toxic and explosive methanol
  • glycerol or glucose as the sole carbon source
  • lower cost of goods through less cooling demand and reduced process time

Please follow this link for more information
VTU´s Pichia pastoris toolbox -
yield enhancing expression platform

 
  • Methanol-induced: up to 25 g/L
  • Methanol-free: up to 20 g/L
  • High diversity of expression tools
  • Sophisticated high-throughput methods
  • Streamlined procedures

Please follow this link for more information
Case Study on the expression of a recombinant bacterial Phytase

Boost yields through fine-tuning of gene expression by increasing genetic diversity

SET UP:

Expression: secreted
  • Methanol-induced expression using VTU´s 1st generation promoter library
  • Methanol-free expression using VTU´s 2nd generation promoter library

Screening size:

  • Small screening - 400 clones applying 2 PAOX1 promoter variants
  • Extended screening - 6000 clones applying 9 PAOX1 promoter variants

Cultivation:

  • The most productive clones were cultivated in 0.75 L bioreactors
  • Pichia strain cultivations were based on simple & cheap chemically defined media
  • No antibiotic markers were used during screening and production


RESULTS:

  • Exceptionally high-yielding Phytase secreting strains
                - 22 g/L - methanol-induced
                - 20 g/L - methanol-free

 
  • A 3.5-fold to 4.6-fold enhancement of protein yields through increasing genetic diversity (increasing VTU´s promoter variants and clone numbers)
Fine tuning of protein expression - get the best expression for each protein

This case study clearly demonstrates that an increase of genetic elements through the application of a higher number of promoter variants from VTU´s promoter libraries significantly boosts protein production in Pichia pastoris, while maintaining competitive development timelines. Through VTU´s pool approach and high-throughput cultivation and screening regime, VTU researchers are able to generate and screen thousands of clones in minimum time.
Timely adjusted co-expression of helper factors provides an additional feature significantly contributing to a substantial reduction of cost of goods in protein manufacturing.
VTU Technology GmbH
Parkring 18, 8074 Grambach, Austria
Phone: +43 (316) 4009-4000
office.technology@vtu.com
www.vtu-technology.com
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