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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Garima Srivastava, Arvind M Kayastha
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cd0a57132dd8406c84ac0c8f070e8b6e
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:cd0a57132dd8406c84ac0c8f070e8b6e
record_format	dspace
spelling	oai:doaj.org-article:cd0a57132dd8406c84ac0c8f070e8b6e2021-11-18T08:32:27ZΒ-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds.1932-620310.1371/journal.pone.0088697https://doaj.org/article/cd0a57132dd8406c84ac0c8f070e8b6e2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24551136/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Fenugreek (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.Garima SrivastavaArvind M KayasthaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 2, p e88697 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Garima Srivastava Arvind M Kayastha Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds.
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.
format	article
author	Garima Srivastava Arvind M Kayastha
author_facet	Garima Srivastava Arvind M Kayastha
author_sort	Garima Srivastava
title	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_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_sort	β-amylase from starchless seeds of trigonella foenum-graecum and its localization in germinating seeds.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/cd0a57132dd8406c84ac0c8f070e8b6e
work_keys_str_mv	AT garimasrivastava bamylasefromstarchlessseedsoftrigonellafoenumgraecumanditslocalizationingerminatingseeds AT arvindmkayastha bamylasefromstarchlessseedsoftrigonellafoenumgraecumanditslocalizationingerminatingseeds
_version_	1718421709176963072

Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds.

Ejemplares similares