Enzymatic degradation of rice bran phytates into myo-inositol phosphates using phytase /
Myo-inositol hexakisphosphate (IP6) or phytate is a naturally occurring polyphosphorylated carbohydrate that is present in substantial amounts in almost all plant and mammalian cells. Partially phosphorylated myo-inositol phosphates have potential applications in many fields including pharmaceutical...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
Kuala Lumpur :
Kulliyyah of Engineering, International Islamic University Malaysia,
2012
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| Subjects: | |
| Online Access: | Click here to view 1st 24 pages of the thesis. Members can view fulltext at the specified PCs in the library. |
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| Summary: | Myo-inositol hexakisphosphate (IP6) or phytate is a naturally occurring polyphosphorylated carbohydrate that is present in substantial amounts in almost all plant and mammalian cells. Partially phosphorylated myo-inositol phosphates have potential applications in many fields including pharmaceutical research and development. Rice bran, a by-product of rice milling was chosen as the source of phytate with up to 4% (w/w) content. Rice bran can be found abundantly in Malaysia at low price. Currently, rice bran is only used to feed livestocks as well as used as fertilizer. The limited exploitation of rice bran prompted this study to try and convert rice bran into myo-inositol phosphates as value added products. Phytases from wildtype (ASUIA 279, ASUIA 271, ASUIA 30) and recombinant strains (M smegmatis, ASUIA 279, PhyFAUIAl) were utilized to degrade rice bran phytate into inorganic phosphates and myo-inositol phosphates. This study investigated the most suitable source of enzyme for rice bran phytate conversion to myo-inositol phosphates as well as optimized the proces~ conditions to achieve high yield of myo-inositol phosphates prior to purification and separation by anion-exchange chromatography. Recombinant M smegmatis phytase gave the highest phosphate (400 micromole phosphates by 2.30U of phytase) and was chosen for the subsequent study. The optimization of process conditions (temperature, agitation, enzyme unit, reaction time) was studied through the application of FCCD from Design Expert software. The model was significant (P-value = 0.0005) with A2 (temperature) having P-value <0.05 and D (enzyme unit), B2 (reaction time), C2 (agitation), D2 (enzyme unit) having p-value<0.01. In addition, good relationship of temperature and agitation could ·be observed through the optimum plot of response surface. Agitation between 190 - 200 rpm and temperature between 50 - 54 °C, gave the optimal phosphate of approximately 140 micromol. The partially purified samples were further purified by HPLC and/or IC and resulted in several compounds such as compunds that having higher myo-inositol phosphates (IP4, IP5, IP6). However, HPIC was not able to separate the compounds under the present experimental conditions. In conclusion, recombinant M. smegmatis phytase obtained from local source has the high potential to produce myo-inositol phosphate as an added-value product from rice bran. |
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| Item Description: | Abstracts in English and Arabic. "A dissertation submitted in fulfilment of the requirement for the degree of Master of Science (Biotechnology Engineering)." --On t.p. |
| Physical Description: | xix, 129 leaves : ill. ; 30cm. |
| Bibliography: | Includes bibliographical references (leaves 89-103). |