Amino acids substitution to improve PhyFAUIA1 phytase activity for animal feed production
Phytate is largely unavailable to monogastric animal such as swine, poultry and fish, as they lack of sufficient endogenous enzymatic activity to hydrolyze phytate. The result is the elimination of precious nutrients that would be beneficial to their growth; furthermore they will excrete most o...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Research Journal of Chemistry and Environment
2010
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/2265/ http://irep.iium.edu.my/2265/ http://irep.iium.edu.my/2265/1/Pytase_Activity_328-333.pdf |
| Summary: | Phytate is largely unavailable to monogastric
animal such as swine, poultry and fish, as they lack of
sufficient endogenous enzymatic activity to hydrolyze
phytate. The result is the elimination of precious
nutrients that would be beneficial to their growth;
furthermore they will excrete most of the indigestive
phytate which can contribute to phosphorus being
over applied to the land. Phosphorus has a beneficial
impact on vegetative growth on land as well as
marine vegetation, causing an increased growth of
weeds.
This enhanced vegetation consumes large
amounts of oxygen, resulting in the loss of aquatic life
and ultimately contributes to water pollution and
eutrophication of ground water and aquatic
environment. Phytase, a type of histidine acid
phosphatase hydrolyzes phytin phosphorus and when
present in an animal's digestive tract, benefits the
animal while reducing total phosphorus levels in
manure. Computer modelling is used to identify and
examine the active site of phytase. The factors
influencing the ligand binding strength in the active
site is analyzed and computational site directed
mutagenesis experiments were carried out to evaluate
the effects of mutations on the binding strength before
and after mutation. From the directive results of computational studies, point mutation was introduced by site directed mutagenesis using polymerase chain reaction (PCR). Mutagenesis is achieved by two step PCR procedure.
Four primers were generated which two of them was
design for carried the mutation at the point of interest
which is complement to each other and the other two
was design for unique restriction site. Several
numbers of single, double as well as triple mutations
has been introduced which will be further
characterized to determine the activity of the enzyme
as compared to the computational results.
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