Restoration of a defective Lactococcus lactis xylose isomerase

Applied and Environmental Microbiology
Joo-Heon Park, C A Batt

Abstract

The genes (xylA) encoding xylose isomerase (XI) from two Lactococcus lactis subsp. lactis strains, 210 (Xyl(-)) and IO-1 (Xyl(+)), were cloned, and the activities of their expressed proteins in recombinant strains of Escherichia coli were investigated. The nucleotide and amino acid sequence homologies between the xylA genes were 98.4 and 98.6%, respectively, and only six amino acid residues differed between the two XIs. The purified IO-1 XI was soluble with K(m) and k(cat) being 2.25 mM and 184/s, respectively, while the 210 XI was insoluble and inactive. Site-directed mutagenesis on 210 xylA showed that a triple mutant possessing R202M/Y218D/V275A mutations regained XI activity and was soluble. The K(m) and k(cat) of this mutant were 4.15 mM and 141/s, respectively. One of the IO-1 XI mutants, S388T, was insoluble and showed negligible activity similar to that of 210 XI. The introduction of a K407E mutation to the IO-1 S388T XI mutant restored its activity and solubility. The dissolution of XI activity in L. lactis subsp. lactis involves a series of mutations that collectively eliminate enzyme activity by reducing the solubility of the enzyme.

References

Aug 1, 1992·The Biochemical Journal·M RangarajanB S Hartley
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·C A BattM A Vandeyar
Jun 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·H L CarrellJ F Biellmann
Jan 1, 1984·Applied and Environmental Microbiology·V B LawlisD J Henner
Apr 15, 1993·The Biochemical Journal·L VarsaniD M Blow
Mar 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·H L CarrellJ P Glusker
Mar 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·H RasmussenM Schülein

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Citations

Apr 22, 2010·BMC Genomics·Yang GuWeihong Jiang
Sep 11, 2013·Bioscience, Biotechnology, and Biochemistry·Mariko Shimizu-KadotaHirofumi Yoshikawa
Feb 2, 2008·Journal of Pharmaceutical Sciences·Saul R TrevinoC Nick Pace
Jun 15, 2010·Journal of Bioscience and Bioengineering·Hyun-Jung KwonDeok-Kun Oh
Nov 12, 2014·Journal of Bacteriology·Benjamin L Golomb, Maria L Marco
Oct 14, 2021·Critical Reviews in Biotechnology·Renan Yuji MiyamotoLeticia Maria Zanphorlin

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