Open AccessExpression, purification, crystallization and preliminary X‐ray analysis of perakine reductase, a new member of the aldo‐keto reductase enzyme superfamily from higher plants
Author(s) -
Rosenthal Cindy,
Mueller Uwe,
Panjikar Santosh,
Sun Lianli,
Ruppert Martin,
Zhao Yu,
Stöckigt Joachim
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910605041x
Subject(s) - aldo keto reductase , aldehyde reductase , reductase , enzyme , escherichia coli , biochemistry , biosynthesis , heterologous expression , chemistry , stereochemistry , aldose reductase , crystallization , biology , gene , organic chemistry , recombinant dna
Perakine reductase (PR) is a novel member of the aldo‐keto reductase enzyme superfamily from higher plants. PR from the plant Rauvolfia serpentina is involved in the biosynthesis of monoterpenoid indole alkaloids by performing NADPH‐dependent reduction of perakine, yielding raucaffrinoline. However, PR can also reduce cinnamic aldehyde and some of its derivatives. After heterologous expression of a triple mutant of PR in Escherichia coli , crystals of the purified and methylated enzyme were obtained by the hanging‐drop vapour‐diffusion technique at 293 K with 100 m M sodium citrate pH 5.6 and 27% PEG 4000 as precipitant. Crystals belong to space group C 222 1 and diffract to 2.0 Å, with unit‐cell parameters a = 58.9, b = 93.0, c = 143.4 Å.