U.S. flagAn official website of the United States government
                    Here's how you know **Official websites use .gov**
                            
                            A **.gov** website belongs to an official government organization in the United States.
 **Secure .gov websites use HTTPS**
                            
                            A **lock** (                            
                              A locked padlock                                                      
                            
                            ) or **https://** means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.


Menu Agricultural Research Service
						
                    U.S. DEPARTMENT OF AGRICULTURE
 close



Research
                Research Home
                            National Programs
                            Research Projects
                            Scientific Manuscripts
                            International Engagement
                            Scientific Software/Models
                            Databases and Datasets
                            Office of Scientific Quality Review
            Media
                News & Features
                            Multimedia
                            Briefing Room
                            Down on the Farm
                            Press Room
                            Science in Your Shopping Cart
                            Scientific Discoveries
                            Tellus
                            Under the Microscope
            About ARS
                About ARS Home
                            Staff Directory
                            Labs and Research Centers (Map)
                            Headquarter Offices
                            Organizational Chart
                            Employee Services (REE Employees Only)
                            Office of Outreach, Diversity, and Equal Opportunity
            Work With Us
                Work With Us Home
                            ARS Vacancies at USAJOBS
                            Careers at ARS Information
                            Scientific Collaborations
                            Extramural Agreements
                            1890 Faculty Research Sabbatical ProgramSearch small
                    Search
            
            ARS Home
                                About ARS
                                Contact Us




ARS Home Research Publications at this Location





- ARS National Programs
- Search for a scientific manuscript
- Search for a research project
- ARS Software
- Programs and Projects
- Collaborations
- Subjects of Investigation
- Publications

Title:   Author  item
                                            Hughes, Stephenitem
                                            STERNER, DAVID - PROGENRA, INC.item
                                            Bischoff, Kennethitem
                                            Hector, Ronald

                                                         - *Ron*item
                                            Dowd, Patrickitem
                                            Qureshi, Nasibitem
                                            BANG, SOOKIE - SDSM&Titem
                                            GRYNAVYSKI, NICOLE - ARRYX, INC.item
                                            CHAKRABARTY, TANIA - ARRYX, INC.item
                                            Johnson, Ericitem
                                            Dien, Bruceitem
                                            Mertens, Jeffreyitem
                                            CAUGHEY, ROBERT - UNIV OF IL COLL OF MEDitem
                                            Liu, Siqingitem
                                            BUTT, TAUSEEF - LIFESENSORS, INC.item
                                            LABAER, JOSHUA - HARVARD INSTITUTEitem
                                            Cotta, Michaelitem
                                            Rich, Joseph
  Submitted to: Plasmid Journal
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 9/5/2008
                                    Publication  Date: 10/30/2008
                                    Citation: Hughes, S.R., Sterner, D.E., Bischoff, K.M., Hector, R.E., Dowd, P.F., Qureshi, N., Bang, S.S., Grynavyski, N., Chakrabarty, T., Johnson, E.T., Dien, B.S., Mertens, J.A., Caughey, R.J., Liu, S., Butt, T.R., Labaer, J., Cotta, M.A., Rich, J.O. 2009. Engineered Saccharomyces cerevisiae strain for improved xylose utilization with a three-plasmid SUMO yeast expression system. Plasmid Journal. 61(1):22-38.Interpretive Summary: Fuel ethanol production from biomass at the industrial level using Saccharomyces cerevisiae shows great promise for satisfying future energy demands, but the limited range of materials that can be fermented remains an obstacle to cost-effective bioethanol production.  Although several genetically engineered strains of S. cerevisiae have been developed that will ferment xylose to ethanol, further optimization is needed.  This 3 vector set of plasmids described used on the automated platform offers the possibility of expressing pentose-utilization enzymes and commercially important peptides and proteins in industrial yeast strains currently used in the production of fuel ethanol from lingnocellulose hydrolysates.
                                Technical Abstract: A three-plasmid yeast expression system utilizing the portable small ubiquitin-like modifier (SUMO) vector set combined with the efficient endogenous yeast protease Ulp1 was developed for production of large amounts of soluble functional protein in Saccharomyces cerevisiae.  Each vector has a different selectable marker (URA, TRP, or LEU), and the system provides high expression levels of three different proteins simultaneously.  This system was adapted for use on a fully automated plasmid-based robotic platform to screen for improved strains of S. cerevisiae engineered to express a peptide of potential commercial value in addition to xylose utilization and metabolic correction enzymes.  First, a novel PCR assembly strategy was used to clone a xylose isomerase (XI) gene into the URA-selectable SUMO vector and the plasmid was placed into the S. cerevisiae strain.  Second, amino acid scanning mutagenesis was used to generate a library of mutagenized genes encoding the bioinsecticidal peptide lycotoxin-1 (Lyt-1) and the library was cloned into the TRP-selectable SUMO vector and placed into the XI-yeast. Third, the Yersinia pestis xylulokinase gene was cloned into the LEU-selectable SUMO vector and placed into the Lyt-1-XI-yeast construct.  Yeast strains expressing XI and xylulokinase with or without Lyt-1 showed improved growth on xylose compared to XI-yeast.


Return to top


Connect with ARS Sign up


ARS Home
                            USDA.gov
                            Plain Writing
                            Policies & Links
                            Civil Rights Statements
                            FOIA
                            Accessibility Statement
                            Privacy Policy
                            Non-Discrimination Statement
                            Quality of Information
                            USA.gov
                            WhiteHouse.gov