Academic Journal
Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae.
| Τίτλος: | Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae. |
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| Συγγραφείς: | Milne N; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., Luttik MAH; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., Cueto Rojas HF; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., Wahl A; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., van Maris AJA; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., Pronk JT; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., Daran JM; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands. Electronic address: j.g.daran@tudelft.nl. |
| Πηγή: | Metabolic engineering [Metab Eng] 2015 Jul; Vol. 30, pp. 130-140. Date of Electronic Publication: 2015 May 30. |
| Τύπος έκδοσης: | Journal Article; Research Support, Non-U.S. Gov't |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Academic Press Country of Publication: Belgium NLM ID: 9815657 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-7184 (Electronic) Linking ISSN: 10967176 NLM ISO Abbreviation: Metab Eng Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Brugge, Belgium ; Orlando, FL : Academic Press, c1999- |
| Ιατρικοί όροι (MeSH): | Saccharomyces cerevisiae*/genetics , Saccharomyces cerevisiae*/metabolism , Gene Expression*, Cation Transport Proteins/*biosynthesis , Nickel/*metabolism , Schizosaccharomyces pombe Proteins/*biosynthesis , Urease/*biosynthesis, Cation Transport Proteins/genetics ; Schizosaccharomyces/genetics ; Schizosaccharomyces/metabolism ; Schizosaccharomyces pombe Proteins/genetics ; Urease/genetics |
| Περίληψη: | In microbial processes for production of proteins, biomass and nitrogen-containing commodity chemicals, ATP requirements for nitrogen assimilation affect product yields on the energy producing substrate. In Saccharomyces cerevisiae, a current host for heterologous protein production and potential platform for production of nitrogen-containing chemicals, uptake and assimilation of ammonium requires 1 ATP per incorporated NH3. Urea assimilation by this yeast is more energy efficient but still requires 0.5 ATP per NH3 produced. To decrease ATP costs for nitrogen assimilation, the S. cerevisiae gene encoding ATP-dependent urease (DUR1,2) was replaced by a Schizosaccharomyces pombe gene encoding ATP-independent urease (ure2), along with its accessory genes ureD, ureF and ureG. Since S. pombe ure2 is a Ni(2+)-dependent enzyme and Saccharomyces cerevisiae does not express native Ni(2+)-dependent enzymes, the S. pombe high-affinity nickel-transporter gene (nic1) was also expressed. Expression of the S. pombe genes into dur1,2Δ S. cerevisiae yielded an in vitro ATP-independent urease activity of 0.44±0.01 µmol min(-1) mg protein(-1) and restored growth on urea as sole nitrogen source. Functional expression of the Nic1 transporter was essential for growth on urea at low Ni(2+) concentrations. The maximum specific growth rates of the engineered strain on urea and ammonium were lower than those of a DUR1,2 reference strain. In glucose-limited chemostat cultures with urea as nitrogen source, the engineered strain exhibited an increased release of ammonia and reduced nitrogen content of the biomass. Our results indicate a new strategy for improving yeast-based production of nitrogen-containing chemicals and demonstrate that Ni(2+)-dependent enzymes can be functionally expressed in S. cerevisiae. (Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.) |
| Contributed Indexing: | Keywords: ATP conservation; ATP-independent urease; Ni-dependent enzyme; Nitrogen metabolism; Physiology; Saccharomyces cerevisiae |
| Substance Nomenclature: | 0 (Cation Transport Proteins) 0 (Nic1 protein, S pombe) 0 (Schizosaccharomyces pombe Proteins) 7OV03QG267 (Nickel) EC 3.5.1.5 (Urease) |
| Entry Date(s): | Date Created: 20150604 Date Completed: 20160422 Latest Revision: 20200930 |
| Update Code: | 20260130 |
| DOI: | 10.1016/j.ymben.2015.05.003 |
| PMID: | 26037463 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1096-7184 |
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| DOI: | 10.1016/j.ymben.2015.05.003 |