PMID: 9654055Jul 8, 1998Paper

Engineering of factors determining alpha-amylase and cyclodextrin glycosyltransferase specificity in the cyclodextrin glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1

European Journal of Biochemistry
R D WindL Dijkhuizen

Abstract

The starch-degrading enzymes alpha-amylase and cyclodextrin glycosyltransferase (CGTase) are functionally and structurally closely related, with CGTases containing two additional domains (called D and E) compared to the three domains of alpha-amylases (A, B and C). Amino acid residue 196 (Thermoanaerobacterium thermosulfurigenes EM1 CGTase numbering) occupies a dominant position in the active-site cleft. All alpha-amylases studied have a small residue at this position (Gly, Leu, Ser, Thr or Val), in contrast to CGTases which have a more bulky aromatic residue (Tyr or Phe) at this position, which is highly conserved. Characterization of the F196G mutant CGTase of T. thermosulfurigenes EM1 revealed that, for unknown reasons, apart from the F196G mutation, domain E as well as a part of domain D had become deleted [mutant F196G(delta'DE)]. This, nevertheless, did not prevent the purification of a stable and active mutant CGTase protein (62 kDa). The mutant protein was more similar to an alpha-amylase protein in terms of the identity of residue 196, and in the domain structure containing, however, some additional C-terminal structure. The mutant showed a strongly reduced temperature optimum. Due to a frameshift mutation in mutant F1...Continue Reading

Citations

Jan 5, 2005·Applied Microbiology and Biotechnology·Qingsheng Qi, Wolfgang Zimmermann
Jun 17, 2008·Applied Biochemistry and Biotechnology·Ivan PishtiyskiBoriana Zhekova
Jan 11, 2001·Biochimica Et Biophysica Acta· van der Veen BAL Dijkhuizen
Mar 21, 2001·Biochimica Et Biophysica Acta·E A MacGregorB Svensson
Jun 17, 2008·Applied and Environmental Microbiology·Juanita Yazmin Damián-AlmazoGloria Saab-Rincón
Jun 29, 2000·Annual Review of Biochemistry·H D Ly, S G Withers
Jul 25, 2000·Protein Engineering·L BeierJ R Cherry
Dec 29, 2006·Chemistry & Biodiversity·Lajos SzenteFriedrich Mandel
Jun 14, 2012·Chemical Biology & Drug Design·Ignatius TintuChittalakkottu Sadasivan
May 20, 2000·Biotechnology & Genetic Engineering Reviews·T Takaha, S M Smith
Sep 27, 2019·Extremophiles : Life Under Extreme Conditions·Carolina Rubiano-LabradorSandra Baena

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