Expression, gene cloning, and characterization of five novel phytases fromfour basidiomycete fungi: Peniophora lycii, Agrocybe pediades, a Ceriporiasp., and Trametes pubescens
Sf. Lassen et al., Expression, gene cloning, and characterization of five novel phytases fromfour basidiomycete fungi: Peniophora lycii, Agrocybe pediades, a Ceriporiasp., and Trametes pubescens, APPL ENVIR, 67(10), 2001, pp. 4701-4707
Phytases catalyze the hydrolysis of phosphomonoester bonds of phytate (myo-
inositol hexakisphosphate), thereby creating lower forms of myo-inositol ph
osphates and inorganic phosphate. In this study, cDNA expression libraries
were constructed from four basidiomycete fungi (Peniophora lycii, Agrocybe
pediades, a Ceriporia sp., and Trametes pubescens) and screened for phytase
activity in yeast. One full-length phytase-encoding cDNA was isolated from
each library, except for the Ceriporia sp. library where two different phy
tase-encoding cDNAs were found. All five phytases were expressed in Aspergi
llus oryzae, purified, and characterized. The phytases revealed temperature
optima between 40 and 60 degreesC and pH optima at 5.0 to 6.0, except for
the P. lycii phytase, which has a pH optimum at 4.0 to 5.0. They exhibited
specific activities in the range of 400 to 1,200 U . mg, of protein(-1) and
were capable of hydrolyzing phytate down to myo-inositol monophosphate. Su
rprisingly, H-1 nuclear magnetic resonance analysis of the hydrolysis of ph
ytate by all five basidiomycete phytases showed a preference for initial at
tack at the 6-phosphate group of phytic acid, a characteristic that was bel
ieved so far not to be seen with fungal phytases. Accordingly, the basidiom
ycete phytases described here should be grouped as 6-phytases (EC 3.1.3.26)
.