Abstract
The yeast Cyberlindnera jadinii has great potential in the biotechnology industry due to its ability to produce a variety of compounds of interest, including carboxylic acids. In this work, we identified genes encoding carboxylate transporters from this yeast species. The functional characterization of sixteen plasma membrane carboxylate transporters belonging to the AceTr, SHS, TDT, MCT, SSS, and DASS families was performed by heterologous expression in Saccharomyces cerevisiae. The newly identified C. jadinii transporters present specificity for mono-, di-, and tricarboxylates. The transporters CjAto5, CjJen6, CjSlc5, and CjSlc13-1 display the broadest substrate specificity; CjAto2 accepts mono- and dicarboxylates; and CjAto1,3,4, CjJen1-5, CjSlc16, and CjSlc13-2 are specific for monocarboxylic acids. A detailed characterization of these transporters, including phylogenetic reconstruction, 3D structure prediction, and molecular docking analysis is presented here. The properties presented by these transporters make them interesting targets to be explored as organic acid exporters in microbial cell factories.
Highlights
The yeast Cyberlindnera jadinii, an Ascomycota belonging to Phaffomycetaceae [1], is an attractive platform for industrial utilization due to its intrinsic robust fermentation features and ability to utilize a wide range of substrates, including hexoses, pentoses, amino acids, and carboxylic acids [2,3,4]
The CBS 1600/NRRL Y-1542 genomes were screened for paralogs of ScAto1 and ScJen1 transporters from S. cerevisiae, resulting in the identification of five ScAto1 (CjAto1-5) and six ScJen1 (CjJen1-6) homologs
This second strategy enabled the detection of a separate set of genes belonging to the families solute/sodium symporter (SSS) (CjSlc5), monocarboxylate transporter (MCT) (CjSlc16), DASS (CjSlc13-1 and CjSlc13-2), and tellurite-resistance/dicarboxylate transporter (TDT) (CjTDT), which are candidates for encoding carboxylate transporters
Summary
The yeast Cyberlindnera jadinii, an Ascomycota belonging to Phaffomycetaceae [1], is an attractive platform for industrial utilization due to its intrinsic robust fermentation features and ability to utilize a wide range of substrates, including hexoses, pentoses, amino acids, and carboxylic acids [2,3,4]. The crystal structures of the bacterial homolog SatP, from Escherichia coli and Citrobacter koseri [12,13], have revealed three conserved hydrophobic residues (F17–Y72–L131), located in the narrowest constriction site, that intimately contribute to the substrate selectivity of these AceTr members [14,15]. Another well-recognized carboxylate transporter family is the tellurite-resistance/dicarboxylate transporter (TDT) (TC 2.A.16), which, as suggested by its nomenclature, is mainly associated with the uptake of dicarboxylates [16]. SLC5 members from the solute/sodium symporter (SSS) family (TC 2.A.21) possess a solute-binding domain associated with the transport of small solutes, such as sugars, vitamins, amino acids, monocarboxylates. and smaller organic ions [22]
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