Dataset Information

Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter.

ABSTRACT:

Background

The recombinant yeast strains displaying the heterologous cellulolytic enzymes on the cell surface using the glycosylphosphatidylinositol (GPI) anchoring system are considered promising biocatalysts for direct conversion of lignocellulosic materials to ethanol. However, the cellulolytic activities of the conventional cellulase-displaying yeast strains are insufficient for the hydrolysis of cellulose. In this study, we constructed novel gene cassettes for the efficient cellulose utilization by cellulase-displaying yeast strains.

Results

The novel gene cassettes for the cell-surface display of Aspergillus aculeatus ?-glucosidase (BGL1) and Trichoderma reeseii endoglucanase II (EGII) were constructed using the promoter and the GPI anchoring region derived from Saccharomyces cerevisiae SED1. The gene cassettes were integrated into the S. cerevisiae genome, then the ?-glucosidase activity of these recombinant strains was evaluated. We revealed that simultaneous utilization of the SED1 promoter and Sed1 anchoring domain in a gene cassette enabled highly-efficient enzyme integration into the cell wall. The ?-glucosidase activity of recombinant yeast cells transduced with the novel gene cassette was 8.4-fold higher than that of a conventional strain. The novel EGII-displaying strain also achieved 106-fold higher hydrolysis activity against the water-insoluble cellulose than a conventional strain. Furthermore, direct ethanol production from hydrothermally processed rice straw was improved by the display of T. reeseii EGII using the novel gene cassette.

Conclusions

We have developed novel gene cassettes for the efficient cell-surface display of exo- and endo-type cellulolytic enzymes. The results suggest that this gene cassette has the wide applicability for cell-surface display and that cellulase-displaying yeasts have significant potential for cost-effective bioethanol production from lignocellulosic biomass.

SUBMITTER: Inokuma K

PROVIDER: S-EPMC3900695 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter.

Inokuma Kentaro K Hasunuma Tomohisa T Kondo Akihiko A

Biotechnology for biofuels 20140114 1

<h4>Background</h4>The recombinant yeast strains displaying the heterologous cellulolytic enzymes on the cell surface using the glycosylphosphatidylinositol (GPI) anchoring system are considered promising biocatalysts for direct conversion of lignocellulosic materials to ethanol. However, the cellulolytic activities of the conventional cellulase-displaying yeast strains are insufficient for the hydrolysis of cellulose. In this study, we constructed novel gene cassettes for the efficient cellulos ...[more]

PMID: 24423072

Similar Datasets

Project description:BACKGROUND:Cellulase is the one of the largest contributors to the high production costs of the lignocellulose-based biorefineries. As the most widely used cellulase producer, Trichoderma reesei has two weaknesses, deficiencies in ?-glucosidase and cellobiohydrolase II. This work aimed at solving this problem by simultaneous enhancement of the beta-exo synergism and exo-exo synergism in T. reesei cellulase to increase the cellulose degrading capability, i.e. enhanced co-expression of the ?-glucosidase gene the cellobiohydrolase II gene of T. reesei. RESULTS:Enhanced co-expression of the ?-glucosidase gene and the cellobiohydrolase II gene in T. reesei using the strong promoter Pcbh1 was found successful in overcoming the two weaknesses. Filter paper activities of T. reesei cellulase were greatly elevated, which were 7.21?±?0.45 (E7, Aabgl1 and Trcbh2) and 7.69?±?0.42 (F6, Anbgl1 and Trcbh2) FPIU/mL. They were much higher than that of the parental strain Rut-C30, 2.45?±?0.36 FPIU/mL. Enzymatic hydrolysis yields were also improved, from 67.22?±?1.61% by Rut-C30 cellulase to 87.98?±?0.65% by E7 cellulase and 86.50?±?1.01% by F6 cellulase. The substrate loading for 1 g glucose release from SECS were decreased, from 2.9637 g SECS using Rut-C30 cellulase to 2.0291 g SECS using E7 cellulase and 2.0573 g SECS using F6 cellulase. As a result, the efficiency of the process from SECS to glucose was substantially improved. CONCLUSIONS:Enhanced co-expression of the ?-glucosidase gene and the cellobiohydrolase II gene in T. reesei using the strong promoter Pcbh1 in T. reesei was proven triumphal in the simultaneous enhancement of the beta-exo synergism and exo-exo synergism in T. reesei cellulase. This strategy also improved the cellulase production, enzymatic hydrolysis yield and the efficiency of the process from SECS to glucose in the context of on-site cellulase production. This work is a commendable attempt in the cellulase composition optimization at the transcriptional level.

Dataset Information

Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter.

Background

Results

Conclusions

Publications

Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets