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Thermostable alpha-galactosidase from Thermotoga neapolitana: cloning, sequencing and expression
M R King1, D A Yernool, D E Eveleigh
1Food Science Department, University of Illinois, Urbana-Champaign 61801, USA.
Abstract:
A gene encoding a thermostable alpha-galactosidase from the hyperthermophile Thermotoga neapolitana was cloned and sequenced. Sequence analysis showed that the 552-amino acid protein is similar to Escherichia coli Raf type alpha-galactosidase and belongs to Family 36 of the glycosyl hydrolases. Recombinant alpha-galactosidase expressed in E. coli has a molecular mass of ca. 61 kDa, and an optimum activity at 93 degrees C at pH 7.0. The enzyme is highly thermostable and retains 75% of activity after heating to 80 degrees C for 4 h. The potential application of the enzyme to high temperature processing of soy molasses has been demonstrated.
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