Functional characterization of a highly specific L-arabinose transporter from Trichoderma reesei.

Havukainen, Sami; Pujol-Giménez, Jonai; Valkonen, Mari; Hediger, Matthias; Landowski, Christopher P (2021). Functional characterization of a highly specific L-arabinose transporter from Trichoderma reesei. Microbial cell factories, 20(1), p. 177. BioMed Central 10.1186/s12934-021-01666-4

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BACKGROUND

Lignocellulose biomass has been investigated as a feedstock for second generation biofuels and other value-added products. Some of the processes for biofuel production utilize cellulases and hemicellulases to convert the lignocellulosic biomass into a range of soluble sugars before fermentation with microorganisms such as yeast Saccharomyces cerevisiae. One of these sugars is L-arabinose, which cannot be utilized naturally by yeast. The first step in L-arabinose catabolism is its transport into the cells, and yeast lacks a specific transporter, which could perform this task.

RESULTS

We identified Trire2_104072 of Trichoderma reesei as a potential L-arabinose transporter based on its expression profile. This transporter was described already in 2007 as D-xylose transporter XLT1. Electrophysiology experiments with Xenopus laevis oocytes and heterologous expression in yeast revealed that Trire2_104072 is a high-affinity L-arabinose symporter with a Km value in the range of [Formula: see text] 0.1-0.2 mM. It can also transport D-xylose but with low affinity (Km [Formula: see text] 9 mM). In yeast, L-arabinose transport was inhibited slightly by D-xylose but not by D-glucose in an assay with fivefold excess of the inhibiting sugar. Comparison with known L-arabinose transporters revealed that the expression of Trire2_104072 enabled yeast to uptake L-arabinose at the highest rate in conditions with low extracellular L-arabinose concentration. Despite the high specificity of Trire2_104072 for L-arabinose, the growth of its T. reesei deletion mutant was only affected at low L-arabinose concentrations.

CONCLUSIONS

Due to its high affinity for L-arabinose and low inhibition by D-glucose or D-xylose, Trire2_104072 could serve as a good candidate for improving the existing pentose-utilizing yeast strains. The discovery of a highly specific L-arabinose transporter also adds to our knowledge of the primary metabolism of T. reesei. The phenotype of the deletion strain suggests the involvement of other transporters in L-arabinose transport in this species.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Forschungsgruppe Nephrologie / Hypertonie
04 Faculty of Medicine > Department of Dermatology, Urology, Rheumatology, Nephrology, Osteoporosis (DURN) > Clinic of Nephrology and Hypertension
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)

UniBE Contributor:

Pujol Gimenez, Jonai, Hediger, Matthias

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1475-2859

Publisher:

BioMed Central

Language:

English

Submitter:

Verena de Serra Frazao-Bill

Date Deposited:

19 Jan 2022 11:23

Last Modified:

05 Dec 2022 15:58

Publisher DOI:

10.1186/s12934-021-01666-4

PubMed ID:

34496831

Uncontrolled Keywords:

Arabinose transporter Pentose fermentation Transmembrane transport Trichoderma reesei

BORIS DOI:

10.48350/162879

URI:

https://boris.unibe.ch/id/eprint/162879

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