Fungal Genomics, Synthetic Biology and Bioprocess Development
The ABF houses the filamentous fungal and non-model yeast synthetic biology capability unique within the government laboratories. This effort also comprises integrated fungal genetic tool and medium scale (15-120L) bioprocess development capability.
Samples for Testing and Learning by a systems biology approach are generated from large shake flask experimental capabilities or through one of our stirred tank reactors that are highly controlled and have advanced monitoring and sampling capabilities, such as:
- Transformation systems and parts (promoters, selection markers, etc.) for eight fungal species.
- Standard modern molecular biology laboratory equipped with all of the thermal cyclers, incubators, electoporators and analytical equipment to generate constructs and verify their insertion sites and DNA sequence.
- Bioreactors with working volumes from 15-120 L for testing genetically engineered strains, for collecting samples from highly controlled and monitored environments and for testing bioprocesses at an intermediate scale.
The limit of filamentous fungal systems generally is the throughput with which genetic changes can be made and tested. Filamentous fungi are less amenable to 96 well culture (1 mL) testing that is translatable back up in scale. Maximum stirred tank reactor volume is 120L. This capability is primarily applicable to one of our three main hosts, Aspergillus pseudoterreus but also is beneficial in working with SNL on the basidiomycete yeast, Rhodosporidium toruloides, and other hosts within current CRADA projects and future ABF users.
References and Additional Information: References/Webpage Links
Yaegashi, J., et al., Rhodosporidium toruloides: a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts. Biotechnol Biofuels, 2017. 10: p. 241.
Dai, Z., et al., Agrobacterium tumefaciens-mediated transformation of oleaginous yeast Lipomyces species. Appl Microbiol Biotechnol, 2017. 101(15): p. 6099-6110.
Dai, Z., et al., Impact of alg3 gene deletion on growth, development, pigment production, protein secretion, and functions of recombinant Trichoderma reesei cellobiohydrolases in Aspergillus niger. Fungal Genet Biol, 2013. 61: p. 120-32.
Pacific Northwest National Laboratory
Sandia National Laboratories