Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres

Background Cyclodextrin glucanotransferase (CGTase) from Amphibacillus sp. NPST-10 was covalently immobilized onto amino-functionalized magnetic double mesoporous core-shell silica nanospheres (mag@d-SiO2@m-SiO2-NH2), and the properties of the immobilized enzyme were investigated. The synthesis proc...

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Autores principales:	Ibrahim,Abdelnasser S.S., Al-Salamah,Ali A, El-Toni,Ahmed Mohamed, El-Tayeb,Mohamed A, Elbadawi,Yahya B
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2014
Materias:	Amphibacillus sp Cyclodextrin glucanotransferase Cyclodextrins Double mesoporous core-shell silica nanospheres Immobilization
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000200001
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spelling	oai:scielo:S0717-345820140002000012014-09-02Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheresIbrahim,Abdelnasser S.S.Al-Salamah,Ali AEl-Toni,Ahmed MohamedEl-Tayeb,Mohamed AElbadawi,Yahya B Amphibacillus sp Cyclodextrin glucanotransferase Cyclodextrins Double mesoporous core-shell silica nanospheres Immobilization Background Cyclodextrin glucanotransferase (CGTase) from Amphibacillus sp. NPST-10 was covalently immobilized onto amino-functionalized magnetic double mesoporous core-shell silica nanospheres (mag@d-SiO2@m-SiO2-NH2), and the properties of the immobilized enzyme were investigated. The synthesis process of the nanospheres included preparing core magnetic magnetite (Fe3O4) nanoparticles, coating the Fe3O4 with a dense silica layer, followed by further coating with functionalized or non-functionalized mesoporous silica shell. The structure of the synthesized nanospheres was characterized using TEM, XRD, and FT-IR analyses. CGTase was immobilized onto the functionalized and non-functionalized nanospheres by covalent attachment and physical adsorption. Results The results indicated that the enzyme immobilization by covalent attachment onto the activated mag@d-SiO2@m-SiO2-NH2, prepared using anionic surfactant, showed highest immobilization yield (98.1%), loading efficiency (96.2%), and loading capacity 58 µg protein [CGTase]/mg [nanoparticles]) which were among the highest yields reported so far for CGTase. Compared with the free enzyme, the immobilized CGTase demonstrated a shift in the optimal temperature from 50°C to 50-55°C, and showed a significant enhancement in the enzyme thermal stability. The optimum pH values for the activity of the free and immobilized CGTase were pH 8 and pH 8.5, respectively, and there was a significant improvement in pH stability of the immobilized enzyme. Moreover, the immobilized CGTase exhibited good operational stability, retaining 56% of the initial activity after reutilizations of ten successive cycles. Conclusion The enhancement of CGTase properties upon immobilization suggested that the applied nano-structured carriers and immobilization protocol are promising approach for industrial bioprocess for production of cyclodextrins using immobilized CGTase.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.17 n.2 20142014-03-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000200001en10.1016/j.ejbt.2014.01.001
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Amphibacillus sp Cyclodextrin glucanotransferase Cyclodextrins Double mesoporous core-shell silica nanospheres Immobilization
spellingShingle	Amphibacillus sp Cyclodextrin glucanotransferase Cyclodextrins Double mesoporous core-shell silica nanospheres Immobilization Ibrahim,Abdelnasser S.S. Al-Salamah,Ali A El-Toni,Ahmed Mohamed El-Tayeb,Mohamed A Elbadawi,Yahya B Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
description	Background Cyclodextrin glucanotransferase (CGTase) from Amphibacillus sp. NPST-10 was covalently immobilized onto amino-functionalized magnetic double mesoporous core-shell silica nanospheres (mag@d-SiO2@m-SiO2-NH2), and the properties of the immobilized enzyme were investigated. The synthesis process of the nanospheres included preparing core magnetic magnetite (Fe3O4) nanoparticles, coating the Fe3O4 with a dense silica layer, followed by further coating with functionalized or non-functionalized mesoporous silica shell. The structure of the synthesized nanospheres was characterized using TEM, XRD, and FT-IR analyses. CGTase was immobilized onto the functionalized and non-functionalized nanospheres by covalent attachment and physical adsorption. Results The results indicated that the enzyme immobilization by covalent attachment onto the activated mag@d-SiO2@m-SiO2-NH2, prepared using anionic surfactant, showed highest immobilization yield (98.1%), loading efficiency (96.2%), and loading capacity 58 µg protein [CGTase]/mg [nanoparticles]) which were among the highest yields reported so far for CGTase. Compared with the free enzyme, the immobilized CGTase demonstrated a shift in the optimal temperature from 50°C to 50-55°C, and showed a significant enhancement in the enzyme thermal stability. The optimum pH values for the activity of the free and immobilized CGTase were pH 8 and pH 8.5, respectively, and there was a significant improvement in pH stability of the immobilized enzyme. Moreover, the immobilized CGTase exhibited good operational stability, retaining 56% of the initial activity after reutilizations of ten successive cycles. Conclusion The enhancement of CGTase properties upon immobilization suggested that the applied nano-structured carriers and immobilization protocol are promising approach for industrial bioprocess for production of cyclodextrins using immobilized CGTase.
author	Ibrahim,Abdelnasser S.S. Al-Salamah,Ali A El-Toni,Ahmed Mohamed El-Tayeb,Mohamed A Elbadawi,Yahya B
author_facet	Ibrahim,Abdelnasser S.S. Al-Salamah,Ali A El-Toni,Ahmed Mohamed El-Tayeb,Mohamed A Elbadawi,Yahya B
author_sort	Ibrahim,Abdelnasser S.S.
title	Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
title_short	Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
title_full	Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
title_fullStr	Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
title_full_unstemmed	Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
title_sort	cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000200001
work_keys_str_mv	AT ibrahimabdelnasserss cyclodextringlucanotransferaseimmobilizationontofunctionalizedmagneticdoublemesoporouscoreshellsilicananospheres AT alsalamahalia cyclodextringlucanotransferaseimmobilizationontofunctionalizedmagneticdoublemesoporouscoreshellsilicananospheres AT eltoniahmedmohamed cyclodextringlucanotransferaseimmobilizationontofunctionalizedmagneticdoublemesoporouscoreshellsilicananospheres AT eltayebmohameda cyclodextringlucanotransferaseimmobilizationontofunctionalizedmagneticdoublemesoporouscoreshellsilicananospheres AT elbadawiyahyab cyclodextringlucanotransferaseimmobilizationontofunctionalizedmagneticdoublemesoporouscoreshellsilicananospheres
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Cyclodextrin glucanotransferase immobilization onto functionalized magnetic double mesoporous core-shell silica nanospheres

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