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The arabinose transporter MtLat-1 is involved in hemicellulase induction in Myceliophthora

ABSTRACT: In this study, we investigated the role of L-arabinose transporter MtLat-1 in the induction of hemicellulase genes of M. thermophila. The deletion of MtLat-1 dramatically improved protein secretion and hemicellulase activities on arabinan. Comparative transcriptomic analysis indicated MtLat-1 repressed expression of hemcellulase genes. Furthermore, based on analyses of non-transporting MtLat-1 mutant, we found MtLat-1 in M. thermophila was involved in arabinose transportation, sensing, and tdownstream signalling cascades. Additionally, we also demonstrated Mtlat-1 expression is directly regulated by transcription factor MtAra-1 and MtAra-1 is involved in L-arabinose transporter and metabolism and arabinan hydrolysis.

ORGANISM(S): Thermothelomyces thermophilus

PROVIDER: GSE221464 | GEO | 2023/03/29

REPOSITORIES: GEO

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Project description:L-Arabinose occurs at economically relevant levels in lignocellulosic hydrolysates. Especially at low concentrations of L-arabinose, its uptake via the Gal2 galactose transporter is an important rate-controlling step in the complete conversion of these feedstocks by engineered, pentose-metabolizing Saccharomyces cerevisiae strains. Chemostat-based transcriptome analysis yielded 16 putative sugar transporter genes in the filamentous fungus Penicillium chrysogenum whose transcript levels were at least three-fold higher in L-arabinose-limited cultures than in glucose-limited and ethanol-limited cultures. Of five genes that showed an over 30-fold higher transcript level in arabinose-grown cultures, only one (Pc20g01790) restored growth on L-arabinose upon expression in an engineered L-arabinose-fermenting S. cerevisiae strain in which GAL2 had been deleted. Sugar-transport assays indicated that Pc20g01790this transporter, designated as PcAraT, encodes functions as a high-affinity (Km = 0.13 mM) L-arabinose-proton symporter that does not transport xylose or glucose. An L-arabinose-metabolizing S. cerevisiae strain co-expressing Pc20g01790PcAraT and GAL2 showed lower residual substrate concentrations in L-arabinose-limited chemostat cultures (4 mg L-1) than a congenic strain in which L-arabinose import exclusively depended on Gal2 (1.8 g L-1). Inhibition of L-arabinose transport by these sugars was less pronounced than observed with Gal2. A hexose-phosphorylation-deficient, L-arabinose-metabolizing S. cerevisiae strain expressing PcAraT Pc20g0190 grew on 20 g L-1 L-arabinose in the presence of 20 g L-1 glucose, which completely inhibited growth on L-arabinose of a congenic strain dependent on L-arabinose transport via Gal2. Its high affinity and specificity for L-L-arabinose, combined with limited sensitivity to inhibition by glucose and D-D-xylose make PcAraT/ Pc20g01790 a valuable transporter gene for application in metabolic engineering strategies aimed at engineering S. cerevisiae strains for efficient conversion of lignocellulosic hydrolysates.

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The arabinose transporter MtLat-1 is involved in hemicellulase induction in Myceliophthora

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