Literature summary for 4.1.2.10 extracted from

Motojima, F.; Nuylert, A.; Asano, Y.
The crystal structure and catalytic mechanism of hydroxynitrile lyase from passion fruit, Passiflora edulis (2018), FEBS J., 285, 313-324 .

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystals of the His6-tagged hydroxynitrile lyase are prepared using vapor diffusion sitting drop method at 20°C. The crystal structures of Passiflora edulis hydroxynitrile lyase and its C-terminal peptide depleted derivative are determined by molecular replacement method using the template structure of a heat stable protein, SP1, from Populus tremula at 2.8 and 1.8 A resolution, respectively. Passiflora edulis hydroxynitrile lyase belongs to dimeric alpha+beta barrel superfamily consisting of a central beta-barrel in the middle of a dimer. The structure of Passiflora edulis hydroxynitrile lyase complexed with (R)-mandelonitrile is also determined. The hydroxyl group of (R)-mandelonitrile forms hydrogen bonds with His8 and Tyr30 in the active site, whereas the nitrile group is oriented toward the carboxyl group of Glu54, unlike other hydroxynitrile lyases, where it interacts with basic residues typically	Passiflora edulis

Crystallization (Comment)

Organism

crystals of the His6-tagged hydroxynitrile lyase are prepared using vapor diffusion sitting drop method at 20°C. The crystal structures of Passiflora edulis hydroxynitrile lyase and its C-terminal peptide depleted derivative are determined by molecular replacement method using the template structure of a heat stable protein, SP1, from Populus tremula at 2.8 and 1.8 A resolution, respectively. Passiflora edulis hydroxynitrile lyase belongs to dimeric alpha+beta barrel superfamily consisting of a central beta-barrel in the middle of a dimer. The structure of Passiflora edulis hydroxynitrile lyase complexed with (R)-mandelonitrile is also determined. The hydroxyl group of (R)-mandelonitrile forms hydrogen bonds with His8 and Tyr30 in the active site, whereas the nitrile group is oriented toward the carboxyl group of Glu54, unlike other hydroxynitrile lyases, where it interacts with basic residues typically

Passiflora edulis

Protein Variants

Protein Variants	Comment	Organism
E54A	completely inactive mutant enzyme	Passiflora edulis
E54Q	mutant enzyme exhibits about 8 of wild-type activity	Passiflora edulis
H8A	completely inactive mutant enzyme	Passiflora edulis
N101A	completely inactive mutant enzyme	Passiflora edulis
N101D	completely inactive mutant enzyme	Passiflora edulis
R56A	mutant enzyme exhibits about 5% of wild-type activity	Passiflora edulis
S66A	mutant enzyme exhibits about 10p% of wild-type activity	Passiflora edulis
Y30F	mutant enzyme exhibits about 5% of wild-type activity	Passiflora edulis

Organism

Organism	UniProt	Comment	Textmining
Passiflora edulis	A0A1L7NZN4	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(R)-mandelonitrile	-	Passiflora edulis	cyanide + benzaldehyde	-	r
cyanide + benzaldehyde	-	Passiflora edulis	(R)-mandelonitrile	-	r

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Passiflora edulis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4.5	-	assay at	Passiflora edulis