Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.

Cyclodextrin glycosyltransferase (CGTase) is an enzyme that produces cyclodextrins (CDs) from starch and related carbohydrates, producing a mixture of α-, β-, and γ-CDs in different amounts. CGTase production, mainly by Bacillus sp., depends on fermentation conditions such as pH, temperature, concen...

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Main Authors: María Belem Arce-Vázquez, Edith Ponce-Alquicira, Ezequiel Delgado-Fornué, Ruth Pedroza-Islas, Gerardo Díaz-Godínez, JORGE SORIANO-SANTOS
Format: Article
Language:English
Published: Frontiers Media S.A. 2016-09-01
Series:Frontiers in Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01513/full
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author María Belem Arce-Vázquez
Edith Ponce-Alquicira
Ezequiel Delgado-Fornué
Ruth Pedroza-Islas
Gerardo Díaz-Godínez
JORGE SORIANO-SANTOS
author_facet María Belem Arce-Vázquez
Edith Ponce-Alquicira
Ezequiel Delgado-Fornué
Ruth Pedroza-Islas
Gerardo Díaz-Godínez
JORGE SORIANO-SANTOS
author_sort María Belem Arce-Vázquez
collection DOAJ
description Cyclodextrin glycosyltransferase (CGTase) is an enzyme that produces cyclodextrins (CDs) from starch and related carbohydrates, producing a mixture of α-, β-, and γ-CDs in different amounts. CGTase production, mainly by Bacillus sp., depends on fermentation conditions such as pH, temperature, concentration of nutrients, carbon and nitrogen sources, among others. Bacillus megaterium CGTase produces those three types of CDs, however, β-CD should prevail. Although waxy corn starch (CS) is used industrially to obtain CGTase and CDs because of its high amylopectin content, alternative sources such as amaranth starch (AS) could be used to accomplish those purposes. AS has high susceptibility to the amylolytic activity of CGTase because of its 80% amylopectin content. Therefore, the aim of this work was evaluate the AS as carbon source for CGTase production by B. megaterium in a submerged fermentation. Afterwards, the CGTase was purified partially and its activity to synthesize α-, β- and γ-CDs was evaluated using 1% AS as substrate. B. megaterium produced a 66 kDa CGTase (Topt=50°C; pHopt=8.0), from the early exponential growth phase which lasted 36 h. The maximum CGTase specific activity (106.62±8.33 U/mg protein) was obtained after 36 h of culture. CGTase obtained with a Km=0.152 mM and a Vmax=13.4 µM/min yielded 40.47% total CDs using AS which was roughly twice as much as that of corn starch (CS; 24.48%). High costs to produce CDs in the pharmaceutical and food industries might be reduced by using AS because of its higher α-, β- and γ-CDs production (12.81%, 17.94% and 9.92%, respectively) in a shorter time than that needed for CS.
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spelling doaj.art-2b42671f6088476688855d70e108a7562022-12-21T18:19:32ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2016-09-01710.3389/fmicb.2016.01513220495Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.María Belem Arce-Vázquez0Edith Ponce-Alquicira1Ezequiel Delgado-Fornué2Ruth Pedroza-Islas3Gerardo Díaz-Godínez4JORGE SORIANO-SANTOS5Metropolitan Autonomus University, Campus IztapalapaMetropolitan Autonomus University, Campus IztapalapaUniversity of GuadalajaraIberoamericana UniversityAutonomous University of TlaxcalaMetropolitan Autonomus University, Campus IztapalapaCyclodextrin glycosyltransferase (CGTase) is an enzyme that produces cyclodextrins (CDs) from starch and related carbohydrates, producing a mixture of α-, β-, and γ-CDs in different amounts. CGTase production, mainly by Bacillus sp., depends on fermentation conditions such as pH, temperature, concentration of nutrients, carbon and nitrogen sources, among others. Bacillus megaterium CGTase produces those three types of CDs, however, β-CD should prevail. Although waxy corn starch (CS) is used industrially to obtain CGTase and CDs because of its high amylopectin content, alternative sources such as amaranth starch (AS) could be used to accomplish those purposes. AS has high susceptibility to the amylolytic activity of CGTase because of its 80% amylopectin content. Therefore, the aim of this work was evaluate the AS as carbon source for CGTase production by B. megaterium in a submerged fermentation. Afterwards, the CGTase was purified partially and its activity to synthesize α-, β- and γ-CDs was evaluated using 1% AS as substrate. B. megaterium produced a 66 kDa CGTase (Topt=50°C; pHopt=8.0), from the early exponential growth phase which lasted 36 h. The maximum CGTase specific activity (106.62±8.33 U/mg protein) was obtained after 36 h of culture. CGTase obtained with a Km=0.152 mM and a Vmax=13.4 µM/min yielded 40.47% total CDs using AS which was roughly twice as much as that of corn starch (CS; 24.48%). High costs to produce CDs in the pharmaceutical and food industries might be reduced by using AS because of its higher α-, β- and γ-CDs production (12.81%, 17.94% and 9.92%, respectively) in a shorter time than that needed for CS.http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01513/fullBacillus megateriumSubmerged fermentationCyclodextrinAmaranth starchCGTase
spellingShingle María Belem Arce-Vázquez
Edith Ponce-Alquicira
Ezequiel Delgado-Fornué
Ruth Pedroza-Islas
Gerardo Díaz-Godínez
JORGE SORIANO-SANTOS
Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.
Frontiers in Microbiology
Bacillus megaterium
Submerged fermentation
Cyclodextrin
Amaranth starch
CGTase
title Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.
title_full Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.
title_fullStr Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.
title_full_unstemmed Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.
title_short Integral use of amaranth starch to obtain cyclodextrin glycosyltransferase, by Bacillus megaterium, to produce β-cyclodextrin.
title_sort integral use of amaranth starch to obtain cyclodextrin glycosyltransferase by bacillus megaterium to produce β cyclodextrin
topic Bacillus megaterium
Submerged fermentation
Cyclodextrin
Amaranth starch
CGTase
url http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01513/full
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