Methanol regulated yeast promoters: production vehicles and toolbox for synthetic biology.

Brigitte Gasser,Diethard Mattanovich,Matthias G Steiger

doi:10.1186/s12934-015-0387-1

Abstract

Promoters are indispensable elements of a standardized parts collection for synthetic biology. Regulated promoters of a wide variety of well-defined induction ratios and expression strengths are highly interesting for many applications. Exemplarily, we discuss the application of published genome scale transcriptomics data for the primary selection of methanol inducible promoters of the yeast Pichia pastoris (Komagataella sp.). Such a promoter collection can serve as an excellent toolbox for cell and metabolic engineering, and for gene expression to produce heterologous proteins.

Highlights

A major task of synthetic biology is the provision of standardized elements for rapid assembly of predictable recombinant gene expression cassettes [1, 2]
There are a plethora of studies which characterize, e.g. constitutive promoters of different strength for Escherichia coli [6], Aspergillus niger [7] or Pichia pastoris [8]
D Induction on methanol was classified based on the transcriptional regulation patterns obtained by [16, 17] by comparing expression levels of cells grown on methanol to cells grown on glucose or glycerol upon induction by methanol, these promoters feature a wide variety of induction degrees, defined as the ratio of expression levels in the induced state vs. the non-induced state

Summary

Background

A major task of synthetic biology is the provision of standardized elements for rapid assembly of predictable recombinant gene expression cassettes [1, 2] These elements include vectors, selection markers, and most importantly collections of regulatory elements like promoters, transcription terminators, secretory leaders and other signal sequences. Komagataella sp.) have gained great interest as production hosts for recombinant proteins [10] and more recently as platform for metabolite production [2] Both applications require promoter collections of different strength for metabolic and cell engineering to enable and enhance productivity. We have redefined the methanol assimilation pathway of P. pastoris [16], a finding that was initially based on the identification of all genes that are upregulated on methanol as a substrate These include hitherto unknown genes, controlled by promoters of a wide range of expression strength on methanol (Table 1).

Methanol inductiond

Conclusions