We develop new technologies for heterologous protein expression in various yeast hosts. Our goal is to develop solutions to various bottlenecks associated with protein expression in yeast that can be implemented at both lab and commercial scales. Our work typically involves working with proteins of commercial interest to NEB or collaborating with researchers outside NEB. We primarily use Kluyveromyces lactis and Pichia pastoris, but also have genetics experience with Schizosaccharomyces and Candida species.
A schematic depicting a typical strategy for secreting heterologous proteins from the yeast Kluyveromyces lactis. In the nucleus, DNA encoding a protein fusion of the α-MF domain (blue) and a desired recombinant protein (black) is integrated into the yeast genome. Expression is driven by the PLAC4-PBI promoter (arrow). Upon expression of the fusion protein, a signal peptide in the α-MF domain directs translocation of the fusion protein into the lumen of the endoplasmic reticulum (ER) and is removed by signal peptidase (SP). Secretory vesicles (circles) transport the fusion protein to the Golgi where the Kex endoprotease cleaves the α-MF pro-domain, releasing a mature form of the desired protein. The protein of interest is then transported via vesicles to the plasma membrane (PM) where it is secreted from the cell.
Taron, C.H., Samuelson, J.C. and Morrison, L. (2020) Microbial expression and purification; one company’s historical perspective. BioProcess Int. 18:9.
Sakhtah, H., Behler, J., Ali-Reynolds, A., Causey, T.B. Vainauskas, S. and Taron, C.H. (2019) A novel regulated hybrid promoter that permits auto-induction of heterologous protein expression in Kluyveromyces lactis. Appl. Environ. Microbiol. PMID: 31053583
Chuzel, L., Ganatra, M.B., Schermerhorn, K.M., Gardner, A.F., Anton, B.P. and Taron, C.H. (2017) Complete genome sequence of Kluyveromyces lactis strain GG799, a common yeast host for heterologous protein expression. Genome Announc. PMID: 28751387
Ganatra, M.B., Vainauskas, S., Hong, J., Taylor, T.E., Denson, J.M., Esposito, D., Read, J.D., Schmeisser, H., Zoon, K.C., Hartley, J.L. and Taron, C.H. (2011) A set of aspartyl protease-deficient strains for improved expression of heterologous proteins in Kluyveromyces lactis. FEMS Yeast Res. PMID: 21166768