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

Kian Mau Goh, Nor Muhammad Mahadi, Osman Hassan, Raja Noor Zaliha Raja Abdul Rahman, Rosli Md Illias

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)118-126
Number of pages9
JournalJournal of Molecular Catalysis B: Enzymatic
Volume49
Issue number1-4
DOIs
Publication statusPublished - 16 Nov 2007

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Manihot
Cyclodextrins
Starch
Enzymes
Buffers
Toluene
Ointments
Viscosity
Glucose
Acetates

Keywords

  • Bacillus sp. G1
  • Cyclodextrin glucanotransferase
  • Gamma-cyclodextrin
  • Pullulanase
  • Site-directed mutagenesis
  • Tapioca starch

ASJC Scopus subject areas

  • Biochemistry
  • Catalysis
  • Process Chemistry and Technology

Cite this

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.

In: Journal of Molecular Catalysis B: Enzymatic, Vol. 49, No. 1-4, 16.11.2007, p. 118-126.

Research output: Contribution to journalArticle

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. In: Journal of Molecular Catalysis B: Enzymatic. 2007 ; Vol. 49, No. 1-4. pp. 118-126.
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AU - Mahadi, Nor Muhammad

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N2 - 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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KW - Pullulanase

KW - Site-directed mutagenesis

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