Change in maltose- and soluble starch-hydrolyzing activities of chimeric α-glucosidases of Mucor javanicus and Aspergillus oryzae

Manabu Sugimoto, Takeshi Ohta, Fusako Kawai

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The chimeric α-glucosidases of Mucor javanicus and Aspergillus oryzae, which has high activity toward not only maltooligosaccharides but also soluble starch and has high activity toward maltooligosaccharides but faint activity toward soluble starch, respectively, were constructed by shuffling the C-terminal regions where low homology is observed between the two enzymes. The chimera genes were expressed in Pichia pastoris to produce and secrete the enzymes that have predicted molecular masses in the culture medium. The two chimeric M. javanicus α-glucosidases, of which the N- and C-terminal regions are substituted for those of A. oryzae, respectively, decreased in soluble starch-hydrolyzing activity, however, increased in maltose-hydrolyzing activity by 2.1 and 4.9 times higher than that of the native form of M. javanicus α-glucosidase, respectively. The chimeric enzymes changed on the Vmax values for maltose significantly, whereas the Km values were similar to that of the native enzyme.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1645
Issue number1
DOIs
Publication statusPublished - Jan 31 2003

Fingerprint

Glucosidases
Aspergillus oryzae
Mucor
Maltose
Aspergillus
Starch
Enzymes
Pichia
Molecular mass
Culture Media
Genes
maltooligosaccharides

Keywords

  • α-Glucosidase
  • Aspergillus oryzae
  • Chimeric enzyme
  • Glycoside hydrolase family 31
  • Mucor javanicus
  • Subsite

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics

Cite this

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abstract = "The chimeric α-glucosidases of Mucor javanicus and Aspergillus oryzae, which has high activity toward not only maltooligosaccharides but also soluble starch and has high activity toward maltooligosaccharides but faint activity toward soluble starch, respectively, were constructed by shuffling the C-terminal regions where low homology is observed between the two enzymes. The chimera genes were expressed in Pichia pastoris to produce and secrete the enzymes that have predicted molecular masses in the culture medium. The two chimeric M. javanicus α-glucosidases, of which the N- and C-terminal regions are substituted for those of A. oryzae, respectively, decreased in soluble starch-hydrolyzing activity, however, increased in maltose-hydrolyzing activity by 2.1 and 4.9 times higher than that of the native form of M. javanicus α-glucosidase, respectively. The chimeric enzymes changed on the Vmax values for maltose significantly, whereas the Km values were similar to that of the native enzyme.",
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AU - Ohta, Takeshi

AU - Kawai, Fusako

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N2 - The chimeric α-glucosidases of Mucor javanicus and Aspergillus oryzae, which has high activity toward not only maltooligosaccharides but also soluble starch and has high activity toward maltooligosaccharides but faint activity toward soluble starch, respectively, were constructed by shuffling the C-terminal regions where low homology is observed between the two enzymes. The chimera genes were expressed in Pichia pastoris to produce and secrete the enzymes that have predicted molecular masses in the culture medium. The two chimeric M. javanicus α-glucosidases, of which the N- and C-terminal regions are substituted for those of A. oryzae, respectively, decreased in soluble starch-hydrolyzing activity, however, increased in maltose-hydrolyzing activity by 2.1 and 4.9 times higher than that of the native form of M. javanicus α-glucosidase, respectively. The chimeric enzymes changed on the Vmax values for maltose significantly, whereas the Km values were similar to that of the native enzyme.

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