Presentation Information
[1ENZ-06]Accelerating Trichoderma reesei strain development with high throughput methods
Norman Adlung1, ○Tino Koponen1, Shaila Eamen1, Ronja Haikarainen1, Gasser Shahin1, Nina Aro1 (1. VTT Technical Research Centre of Finland (Finland))
Keywords:
Biofoundry,High throughput,Laccase,Trichoderma reesei,Screening
Efficient, consistent, and thorough experimenting helps us to engineer complex systems such as non-model microbial organisms. Advanced biofoundry tools, for instance liquid handling robots and automated fermentation systems, allow screening of culture conditions and genotypes that have the potential to boost industrial biotechnology processes. However, the high-throughput methods are underdeveloped, which hinders progress, for example when engineering filamentous fungi. Trichoderma reesei is by nature an efficient protein secretor and widely used already today for production of homologous as well as heterologous proteins. Automation can significantly speed up development of T. reesei strain development and thereby advance its use as an industrial production host.
We have developed a semi-automated high throughput transformant screening workflow for T. reesei. Manual low-throughput unit operations such as colony picking, spore harvesting and microtiter well screening are now executed with liquid handling robots that increase the throughput. As a test case, we explored a recombinant protein over-expression library with laccase-secreting T. reesei strains. As a result of the small-scale cultivations, we were able to identify a top-performing overexpression variant. The same variant had a two-fold increase in laccase expression in bioreactor fermentation, which highlights the potential of our workflow for screening T. reesei strains to be used in bioproduction at scale.
We have developed a semi-automated high throughput transformant screening workflow for T. reesei. Manual low-throughput unit operations such as colony picking, spore harvesting and microtiter well screening are now executed with liquid handling robots that increase the throughput. As a test case, we explored a recombinant protein over-expression library with laccase-secreting T. reesei strains. As a result of the small-scale cultivations, we were able to identify a top-performing overexpression variant. The same variant had a two-fold increase in laccase expression in bioreactor fermentation, which highlights the potential of our workflow for screening T. reesei strains to be used in bioproduction at scale.
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