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- Title
Molecular cloning and the biochemical characterization of two novel phytases from B. subtilis 168 and B. licheniformis.
- Authors
Tye, A J; Siu, F K Y; Leung, T Y C; Lim, B L
- Abstract
A novel phytase gene ( phyL) was cloned from Bacillus licheniformis by multiple steps of degenerate and inverse PCR. The coding region of the phyL gene was 1,146 bp in size and a promoter region of approximately 300 bp was identified at the upstream sequence. This gene, together with a phytase gene ( 168phyA) identified in the B. subtilis strain 168 genome by a homology search, was cloned and over-expressed in B. subtilis using a phi105MU331 prophage vector system. Up to 35 units of phytase/ml were secreted into the culture media; and mature enzymes of around 44-47 kDa were purified for characterization. Both phytases exhibited broad temperature and pH optima and showed high thermostability. Of the two, the phytase encoded by phyL exhibited higher thermostability, even at a lower calcium concentration, as it was able to recover 80% of its original activity after denaturation at 95 degrees C for 10 min. With their neutral pH optima and good temperature stabilities, these Bacillus phytases are good candidates for animal feed applications and transgenic studies.
- Publication
Applied microbiology and biotechnology, 2002, Vol 59, Issue 2-3, p190
- ISSN
0175-7598
- Publication type
Journal Article
- DOI
10.1007/s00253-002-1033-5