Abstract: Protein kinases participate in a myriad of cellular
processes of major biomedical interest. The in vivo substrate
specificity of these enzymes is a process determined by several
factors, and despite several years of research on the topic, is still
far from being totally understood. In the present work, we have
quantified the contributions to the kinase substrate specificity of
i) the phosphorylation sites and their surrounding residues in the
sequence and of ii) the association of kinases to adaptor or scaffold
proteins. We have used position-specific scoring matrices (PSSMs),
to represent the stretches of sequences phosphorylated by 93 families
of kinases. We have found negative correlations between the number
of sequences from which a PSSM is generated and the statistical
significance and the performance of that PSSM. Using a subset
of 22 statistically significant PSSMs, we have identified specificity
determinant residues (SDRs) for 86% of the corresponding kinase
families. Our results suggest that different SDRs can function as
positive or negative elements of substrate recognition by the different
families of kinases. Additionally, we have found that human proteins
with known function as adaptors or scaffolds (kAS) tend to interact
with a significantly large fraction of the substrates of the kinases to
which they associate. Based on this characteristic we have identified
a set of 279 potential adaptors/scaffolds (pAS) for human kinases,
which is enriched in Pfam domains and functional terms tightly
related to the proposed function. Moreover, our results show that
for 74.6% of the kinase–pAS association found, the pAS colocalize
with the substrates of the kinases they are associated to. Finally, we
have found evidence suggesting that the association of kinases to
adaptors and scaffolds, may contribute significantly to diminish the
in vivo substrate crossed-specificity of protein kinases. In general, our
results indicate the relevance of several SDRs for both the positive
and negative selection of phosphorylation sites by kinase families and
also suggest that the association of kinases to pAS proteins may be
an important factor for the localization of the enzymes with their set
of substrates.
Abstract: Phytases are acid phosphatase enzymes, which
efficiently cleave phosphate moieties from phytic acid, thereby
generating myo-inositol and inorganic phosphate. Thirty four
isolates of endophytic fungi to produce of phytases were isolated
from leaf, stem and root fragments of soybean. Screening of 34
isolates of endophytic fungi identified the phytases produced by
Rhizoctonia sp. and Fusarium verticillioides . The phytase
production were the best induced by phytic acid and rice bran
compared the others inducer in submerged fermentation medium
used. The phytase produced by both Rhizoctonia sp. and F.
verticillioides have pH optimum at 4.0 and 5.0 respectively. The
characterization of phytase from Fusarium verticillioides showed that
temperature optimum was 500C and stability until 600C, the pH
optimum 5.0 and pH stability was 2.5 – 6.0, and substrate specificity
were rice bran>soybean meal>corn> coconut cake, respectively.