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Expression System with Chaperones

products-servicesARTES Biotechnology GmbH
June 13th 2012

ARTES Biotechnology offers innovative technologies in the area of yeast expression system with chaperones.

An expression system is a system specifically designed for the production of a specific gene product. It has its individual features and benefits; an ideal expression system depends on the characteristics of the individual gene and protein.

We offer the patented chaperone platform used to increase secretion levels in yeast, preferably in our established high-yield Hansenula (syn. Pichia angusta).

Hansenula Scientific Features:
Genetics:

• High stability of strains up to 800 generations tested
• Co-expression up to 3 different genes
• Multi-copy strains with up to 120 copies per cell
• Expression vectors without antibiotic resistance markers available
• Stable, high yields

Fermentation:
Hansenula cultivated in simple inexpensive fermentation environment with a range of cultivation conditions at 20-45 centigrade,pH 2.5 – 6.0 .The media used is methanol free and inexpensive. Fermentation time span is short, about 70 – 120hrs.

Product:
The Hansenula expression system is cost effective and easy processing. The host protein impurities during secretion are low and the product yield is high, around 13.5g/l. The protein obtained is highly stable in the fermentation.

Reliability:
An established technology and patent protected. It has a high batch to batch reproducibility. The combined effect of gene dosage and promoter efficiency makes the Hansenula expression yields superior to other microbial system.

Hansenula polymorpha platform’s commercial track record:
• Pharmaceutical and feed/food products
• Industrial enzymes approved as food ingredients, a.o. US-FDA
• Applied in contract R&D for 20 years
• Hepatitis B vaccine approved by WHO
• Insulin, interferon alpa2a and Hirudin as therapeutic products
• Genome sequence
• Phytase with EFSA approval for feed enzyme

Hansenula User Reference:
The service has been provided to renowned pharmaceuticals and food industries, such as:
• DSM Nutritional products
• PROTEO Biotech AG
• DuPont Danisco
• Novozyme
• Crucell
• Cadila
• Zydus.

Chaperones – Tool to Improve Secretion
ARTES has developed a genetically manipulated process to optimize a given yeast production strain in order to enhance its productivity.

In order for commercial production of the target protein to be cost effective it is important that the heterologous expression of the protein has the ability to achieve high yields. The technology encompasses the introduction of additional gene-copies of yeast Calnexin into the receiver’s genome. This specific research setup was evaluated with more than ten totally diverse recombinant proteins of commercial interest in shake flasks as well as in pilot fermentation scale. ARTES achieved two- to three fold boost of productivity on proteins already secreted at g/L level before.

The main function of Calnexin is assisting protein folding and quality control, ensuring that only properly folded and assembled proteins proceed further along the secretory pathway. Calnexin retains unfolded or glycoproteins in the ER. Differentiating the gene doses of Calnexin in yeasts may represent a core adjusting screw to improve the capability of host cells to produce foreign proteins.

The amazing result of this particular manipulation strategy is the opportunity to control the yeast product strain which already yields high productivity as well as being able to apply this to totally different recombinant proteins being expressed from an approved yeast expression platform. ARTES has invented a tool and made it possible to take it from small lab scale to large industrial scale fermentation.

Hansenula is an ideal host for combinatory optimization approaches. The research on secretion stimulation effects in Hansenula in particular and other general yeasts is being carried out in ARTES’ labs with work on other chaperones.

ARTES also offers SEB another exclusive chaperone which is involved in increasing secretion in yeast.

For more information or to discuss expression systems with chaperones please contact us directly.

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