Overproduction of an acetylxylan esterase from the extreme thermophile "Caldocellum saccharolyticum" in Escherichia coli

Appl Microbiol Biotechnol. 1990 Nov;34(2):214-9. doi: 10.1007/BF00166783.

Abstract

The xynC gene coding for an acetylxylan esterase from the extreme thermophile "Caldocellum saccharolyticum" was overexpressed in Escherichia coli strain RR28 by cloning the gene downstream from the lacZ promoter region of pUC18 (pNZ1447) or downstream from the temperature-inducible lambda pRpL promoters of pJLA602 (pNZ1600). The protein formed high molecular weight aggregates in induced cells of RR28/pNZ1600 but not in RR28/pNZ1447. The enzyme constituted up to 10% of the total cell protein and was located in the cytoplasmic fraction of RR28/pNZ1447. The acetyl esterase was most active at pH 6.0 and 70-75 degrees C with a half-life of 64 h at 70 degrees C and 30 h at 80 degrees C, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylesterase / biosynthesis*
  • Acetylesterase / genetics
  • Bacteria, Anaerobic / enzymology*
  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Gene Expression Regulation, Enzymologic / genetics
  • Hydrogen-Ion Concentration
  • Plasmids / genetics
  • Temperature

Substances

  • Bacterial Proteins
  • Acetylesterase