TY  - JOUR
AU  - Guterl, K.
AU  - Andexer, J.N.
AU  - Sehl, T.
AU  - von Langermann, J.
AU  - Frindi-Wosch, I.
AU  - Rosenkranz, T.
AU  - Fitter, J.
AU  - Gruber, K.
AU  - Kragl, U.
AU  - Eggert, T.
AU  - Pohl, M.
TI  - Uneven Twins: Comparison of two enantiocomplementary hydroxynitrile lyases with  a/ß--hydrolase fold
JO  - Journal of biotechnology
VL  - 141
SN  - 0168-1656
CY  - Amsterdam [u.a.]
PB  - Elsevier Science
M1  - PreJuSER-4648
PY  - 2009
N1  - The authors thank Julich Chiral Solutions/Codexis for providing MeHNL-DNA and MeHNL-Cys81Ala expressing E. coli cells, Astrid Wirtz for HPLC measurements and Sabine Kruschinski for technical assistance. This work was partially supported by the BMBF in frame of project "Biokatalytische Hydrocyanierung & Hydroformylierung (BioHydroForm) FKZ 0313402C" and by the Deutsche Forschungsgemeinschaft in frame of the research training group "BioNoCo" GK 1166.
AB  - Hydroxynitrile lyases (HNLs) are applied in technical processes for the synthesis of chiral cyanohydrins. Here we describe the thorough characterization of the recently discovered R-hydroxynitrile lyase from Arabidopsis thaliana and its S-selective counterpart from Manihot esculenta (MeHNL) concerning their properties relevant for technical applications. The results are compared to available data of the structurally related S-HNL from Hevea brasiliensis (HbHNL), which is frequently applied in technical processes. Whereas substrate ranges are highly similar for all three enzymes, the stability of MeHNL with respect to higher temperature and low pH-values is superior to the other HNLs with alpha/beta-hydrolase fold. This enhanced stability is supposed to be due to the ability of MeHNL to form tetramers in solution, while HbHNL and AtHNL are dimers. The different inactivation pathways, deduced by means of circular dichroism, tryptophan fluorescence and static light scattering further support these results. Our data suggest different possibilities to stabilize MeHNL and AtHNL for technical applications: whereas the application of crude cell extracts is appropriate for MeHNL, AtHNL is stabilized by addition of polyols. In addition, the molecular reason for the inhibition of MeHNL and HbHNL by acetate could be elucidated, whereas no such inhibition was observed with AtHNL.
KW  - Acetonitriles: metabolism
KW  - Aldehyde-Lyases: chemistry
KW  - Aldehyde-Lyases: genetics
KW  - Aldehyde-Lyases: metabolism
KW  - Amino Acid Sequence
KW  - Arabidopsis: enzymology
KW  - Arabidopsis: genetics
KW  - Enzyme Stability
KW  - Escherichia coli: genetics
KW  - Hevea: enzymology
KW  - Hevea: genetics
KW  - Hydrogen-Ion Concentration
KW  - Hydrolases: genetics
KW  - Manihot: enzymology
KW  - Manihot: genetics
KW  - Molecular Sequence Data
KW  - Plant Proteins: genetics
KW  - Plant Proteins: metabolism
KW  - Recombinant Proteins: genetics
KW  - Recombinant Proteins: metabolism
KW  - Stereoisomerism
KW  - Substrate Specificity
KW  - Temperature
KW  - Time Factors
KW  - Acetonitriles (NLM Chemicals)
KW  - Plant Proteins (NLM Chemicals)
KW  - Recombinant Proteins (NLM Chemicals)
KW  - mandelonitrile (NLM Chemicals)
KW  - Hydrolases (NLM Chemicals)
KW  - Aldehyde-Lyases (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:19433222
UR  - <Go to ISI:>//WOS:000266672600012
DO  - DOI:10.1016/j.jbiotec.2009.03.010
UR  - https://juser.fz-juelich.de/record/4648
ER  -