The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase

A novel mutant enzyme namely H43T CGTase can produce up to 39% γ-cyclodextrin (γ-CD) compared to the native enzyme which produces only 10% γ-CD. The effect of the reaction conditions on γ-CD production was studied using this mutant CGTase. The effects of substrate-buffer combination, starch pret...

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Main Authors: Goh, Kian Mau, Mahadi, Nor Muhammad, Hassan, Osman, Abdul Rahman, Raja Noor Zaliha Raja, Illias, Rosli Md
Format: Article
Published: Elsevier B.V. 2007
Subjects:
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author Goh, Kian Mau
Mahadi, Nor Muhammad
Hassan, Osman
Abdul Rahman, Raja Noor Zaliha Raja
Illias, Rosli Md
author_facet Goh, Kian Mau
Mahadi, Nor Muhammad
Hassan, Osman
Abdul Rahman, Raja Noor Zaliha Raja
Illias, Rosli Md
author_sort Goh, Kian Mau
collection ePrints
description A novel mutant enzyme namely H43T CGTase can produce up to 39% γ-cyclodextrin (γ-CD) compared to the native enzyme which produces only 10% γ-CD. The effect of the reaction conditions on γ-CD production was studied using this mutant CGTase. The effects of substrate-buffer combination, starch pretreatment and concentration, pH, additives and finally the use of a debranching enzyme improved the γ-CD ratio further. The tapioca-acetate pair gave the highest conversion (16% conversion) among four types of starch and four buffer system combinations. Gelatinized starch was preferred compared to raw tapioca starch in producing a high percentage of γ-CD and conversion rate. Higher pH especially pH 8-9 led to a higher proportion of γ-CD, and was relatively more apparent when the concentration of starch was increased. Forty-six percent γ-CD was produced using 2.5% gelatinized tapioca starch at pH 8. Pullulanase enzyme was found to be useful in reducing the viscosity of tapioca starch paste thus increasing the efficiency of utilization of starch by CGTase by at least 20- to 30-fold. Up to 48% γ-CD can be produced when 4% pullulanase-pretreated tapioca starch was reacted with the CGTase mutant. It was also found that the supplementation of the reaction mixture with glucose, toluene, or cyclododecanone improved the γ-CD yield by 42.2, 46.4, 43.4, and 43.4%, respectively. All the parameters involved have been shown to affect the product specificity of the mutant H43T CGTase transglycosylation mechanism.
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spelling utm.eprints-72462009-01-01T04:39:39Z http://eprints.utm.my/7246/ The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase Goh, Kian Mau Mahadi, Nor Muhammad Hassan, Osman Abdul Rahman, Raja Noor Zaliha Raja Illias, Rosli Md TP Chemical technology A novel mutant enzyme namely H43T CGTase can produce up to 39% γ-cyclodextrin (γ-CD) compared to the native enzyme which produces only 10% γ-CD. The effect of the reaction conditions on γ-CD production was studied using this mutant CGTase. The effects of substrate-buffer combination, starch pretreatment and concentration, pH, additives and finally the use of a debranching enzyme improved the γ-CD ratio further. The tapioca-acetate pair gave the highest conversion (16% conversion) among four types of starch and four buffer system combinations. Gelatinized starch was preferred compared to raw tapioca starch in producing a high percentage of γ-CD and conversion rate. Higher pH especially pH 8-9 led to a higher proportion of γ-CD, and was relatively more apparent when the concentration of starch was increased. Forty-six percent γ-CD was produced using 2.5% gelatinized tapioca starch at pH 8. Pullulanase enzyme was found to be useful in reducing the viscosity of tapioca starch paste thus increasing the efficiency of utilization of starch by CGTase by at least 20- to 30-fold. Up to 48% γ-CD can be produced when 4% pullulanase-pretreated tapioca starch was reacted with the CGTase mutant. It was also found that the supplementation of the reaction mixture with glucose, toluene, or cyclododecanone improved the γ-CD yield by 42.2, 46.4, 43.4, and 43.4%, respectively. All the parameters involved have been shown to affect the product specificity of the mutant H43T CGTase transglycosylation mechanism. Elsevier B.V. 2007-11-16 Article PeerReviewed Goh, Kian Mau and Mahadi, Nor Muhammad and Hassan, Osman and Abdul Rahman, Raja Noor Zaliha Raja and Illias, Rosli Md (2007) The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase. Journal of Molecular Catalysis B: Enzymatic, 49 (1-4). p. 118. ISSN 1381-1177 http://dx.doi.org/10.1016/j.molcatb.2007.09.011 10.1016/j.molcatb.2007.09.011
spellingShingle TP Chemical technology
Goh, Kian Mau
Mahadi, Nor Muhammad
Hassan, Osman
Abdul Rahman, Raja Noor Zaliha Raja
Illias, Rosli Md
The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase
title The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase
title_full The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase
title_fullStr The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase
title_full_unstemmed The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase
title_short The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase
title_sort effects of reaction conditions on the production of i³ cyclodextrin from tapioca starch by using a novel recombinant engineered cgtase
topic TP Chemical technology
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