4BIF

Biochemical and structural characterisation of a novel manganese- dependent hydroxynitrile lyase from bacteria

4BIF の概要

• エントリーDOI: 10.2210/pdb4bif/pdb
• 分子名称: CUPIN 2 CONSERVED BARREL DOMAIN PROTEIN, MANGANESE (II) ION (3 entities in total)
• 機能のキーワード: lyase, metal-dependent, mandelonitrile, site-directed mutagenesis
• 由来する生物種: GRANULICELLA TUNDRICOLA
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 139401.58

構造登録者

Hajnal, I.,Lyskowski, A.,Hanefeld, U.,Gruber, K.,Schwab, H.,Steiner, K. (登録日: 2013-04-10, 公開日: 2013-09-11, 最終更新日: 2023-12-20)

主引用文献

Hajnal, I.,Lyskowski, A.,Hanefeld, U.,Gruber, K.,Schwab, H.,Steiner, K.
Biochemical and Structural Characterisation of a Novel Bacterial Manganese-Dependent Hydroxynitrile Lyase.
FEBS J., 280:5815-, 2013

PubMed Abstract: Hydroxynitrile lyases (HNLs), which catalyse the decomposition of cyanohydrins, are found mainly in plants. In vitro, they are able to catalyse the synthesis of enantiopure cyanohydrins, which are versatile building blocks in the chemical industry. Recently, HNLs have also been discovered in bacteria. Here, we report on the detailed biochemical and structural characterization of a hydroxynitrile lyase from Granulicella tundricola (GtHNL), which was successfully heterologously expressed in Escherichia coli. The crystal structure was solved at a crystallographic resolution of 2.5 Å and exhibits a cupin fold. As GtHNL does not show any sequence or structural similarity to any other HNL and does not contain conserved motifs typical of HNLs, cupins represent a new class of HNLs. GtHNL is metal-dependent, as confirmed by inductively coupled plasma/optical emission spectroscopy, and in the crystal structure, manganese is bound to three histidine and one glutamine residue. GtHNL displayed a specific activity of 1.74 U·mg(-1) at pH 6 with (R)-mandelonitrile, and synthesized (R)-mandelonitrile with 90% enantiomeric excess at 80% conversion using 0.5 m benzaldehyde in a biphasic reaction system with methyl tertiary butyl ether.

PubMed: 23981508
DOI: 10.1111/FEBS.12501

実験手法

X-RAY DIFFRACTION (2.46 Å)