Heterologous expression of D-xylulokinase from Pichia stipitis enables high levels of xylitol production by engineered Escherichia coli growing on xylose

Metabolic Engineering
Olubolaji Akinterinwa, Patrick C Cirino

Abstract

Deletion of the Escherichia coli xylulokinase gene (xylB) is essential for achieving high xylitol titers from xylitol-producing E. coli strains growing on glucose in the presence of xylose. Our study suggests that this is due to XylB-catalyzed toxic synthesis of xylitol-phosphate. This activity prohibits the use of xylose as the sole carbon source during xylitol production by E. coli. To overcome this limitation we turned to the yeast Pichia stipitis, which naturally produces xylitol, as a source of xylulokinase (Xyl3). We examined the effects of plasmid-based expression of Xyl3 versus XylB on growth and xylitol production by engineered E. coli strains. Xylulokinase activity assays show similar levels of functional expression of both enzymes (determined as activity on xylulose), and reveal significantly more activity on xylitol by XylB compared to Xyl3. (31)P NMR confirms the production of xylitol-phosphate from in vitro reactions with XylB. Lastly, the replacement of xylB with XYL3 results in drastically enhanced xylitol titers from E. coli strains co-expressing xylose reductase during growth on xylose.

References

Mar 15, 1979·Journal of Molecular Evolution·G A Scangos, A M Reiner
Jul 1, 1988·Plasmid·P E Stevis, N W Ho
Feb 28, 1998·British Dental Journal·W M Edgar
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Mar 2, 2002·Applied and Environmental Microbiology·Yong-Su JinThomas W Jeffries
Jan 31, 2003·Proceedings of the National Academy of Sciences of the United States of America·T B CauseyL O Ingram
Dec 22, 2004·Journal of Applied Microbiology·S BuR Z Fu
Jul 14, 2006·Biotechnology and Bioengineering·Patrick C CirinoLonnie O Ingram
Nov 25, 2006·Journal of Molecular Biology·Eric Di LuccioDavid K Wilson
Jan 12, 2007·Applied Microbiology and Biotechnology·Tom Birger GranströmMatti Leisola

❮ Previous
Next ❯

Citations

May 20, 2014·Applied Microbiology and Biotechnology·Kori L Dunn, Christopher V Rao
Nov 26, 2010·Applied and Environmental Microbiology·Olubolaji Akinterinwa, Patrick C Cirino
Jul 2, 2011·Biotechnology Letters·Manoj Agrawal, Rachel Ruizhen Chen
Dec 17, 2009·Applied Microbiology and Biotechnology·Miho SasakiHideaki Yukawa
May 8, 2010·Metabolic Engineering·Nikhil U Nair, Huimin Zhao
Mar 16, 2011·Biotechnology and Bioengineering·Manoj AgrawalRachel Ruizhen Chen
Jun 20, 2012·Metabolic Engineering·Mervi ToivariMarilyn G Wiebe
Apr 24, 2016·Applied and Environmental Microbiology·Ramanan SekarThomas J DiChristina
Jan 15, 2014·Applied Microbiology and Biotechnology·Hairong ChengZixin Deng
Oct 29, 2013·International Journal of Food Sciences and Nutrition·Hansa Jain, Sanjyot Mulay
May 19, 2019·Applied Microbiology and Biotechnology·Yirong XuHairong Cheng
Sep 22, 2018·Biotechnology for Biofuels·Alexandra Merkx-JacquesDavid L Woodhall
Feb 11, 2021·Critical Reviews in Biotechnology·Angelo B BañaresWook-Jin Chung
Dec 14, 2011·ACS Chemical Biology·Shan-He ChenChi-Huey Wong

❮ Previous
Next ❯

Related Concepts

Related Feeds

Aminoglycosides (ASM)

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Aminoglycosides

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.