Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds.
Fenugreek (Trigonella foenum-graecum) seeds do not contain starch as carbohydrate reserve. Synthesis of starch is initiated after germination. A β-amylase from ungerminated fenugreek seeds was purified to apparent electrophoretic homogeneity. The enzyme was purified 210 fold with specific activity o...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24551136/pdf/?tool=EBI |
| _version_ | 1818337599048122368 |
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| author | Garima Srivastava Arvind M Kayastha |
| author_facet | Garima Srivastava Arvind M Kayastha |
| author_sort | Garima Srivastava |
| collection | DOAJ |
| description | Fenugreek (Trigonella foenum-graecum) seeds do not contain starch as carbohydrate reserve. Synthesis of starch is initiated after germination. A β-amylase from ungerminated fenugreek seeds was purified to apparent electrophoretic homogeneity. The enzyme was purified 210 fold with specific activity of 732.59 units/mg. Mr of the denatured enzyme as determined from SDS-PAGE was 58 kD while that of native enzyme calculated from size exclusion chromatography was 56 kD. Furthermore, its identity was confirmed to be β-amylase from MALDI-TOF analysis. The optimum pH and temperature was found to be 5.0 and 50°C, respectively. Starch was hydrolyzed at highest rate and enzyme showed a Km of 1.58 mg/mL with it. Antibodies against purified Fenugreek β-amylase were generated in rabbits. These antibodies were used for localization of enzyme in the cotyledon during different stages of germination using fluorescence and confocal microscopy. Fenugreek β-amylase was found to be the major starch degrading enzyme depending on the high amount of enzyme present as compared to α-amylase and also its localization at the periphery of amyloplasts. A new finding in terms of its association with protophloem was observed. Thus, this enzyme appears to be important for germination of seeds. |
| first_indexed | 2024-12-13T14:57:46Z |
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| id | doaj.art-cd0a57132dd8406c84ac0c8f070e8b6e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T14:57:46Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-cd0a57132dd8406c84ac0c8f070e8b6e2022-12-21T23:41:12ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0192e8869710.1371/journal.pone.0088697Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds.Garima SrivastavaArvind M KayasthaFenugreek (Trigonella foenum-graecum) seeds do not contain starch as carbohydrate reserve. Synthesis of starch is initiated after germination. A β-amylase from ungerminated fenugreek seeds was purified to apparent electrophoretic homogeneity. The enzyme was purified 210 fold with specific activity of 732.59 units/mg. Mr of the denatured enzyme as determined from SDS-PAGE was 58 kD while that of native enzyme calculated from size exclusion chromatography was 56 kD. Furthermore, its identity was confirmed to be β-amylase from MALDI-TOF analysis. The optimum pH and temperature was found to be 5.0 and 50°C, respectively. Starch was hydrolyzed at highest rate and enzyme showed a Km of 1.58 mg/mL with it. Antibodies against purified Fenugreek β-amylase were generated in rabbits. These antibodies were used for localization of enzyme in the cotyledon during different stages of germination using fluorescence and confocal microscopy. Fenugreek β-amylase was found to be the major starch degrading enzyme depending on the high amount of enzyme present as compared to α-amylase and also its localization at the periphery of amyloplasts. A new finding in terms of its association with protophloem was observed. Thus, this enzyme appears to be important for germination of seeds.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24551136/pdf/?tool=EBI |
| spellingShingle | Garima Srivastava Arvind M Kayastha Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. PLoS ONE |
| title | Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. |
| title_full | Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. |
| title_fullStr | Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. |
| title_full_unstemmed | Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. |
| title_short | Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. |
| title_sort | β amylase from starchless seeds of trigonella foenum graecum and its localization in germinating seeds |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24551136/pdf/?tool=EBI |
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