Modulation of transglycosylation and improved malto-oligosaccharide synthesis by protein engineering of maltogenic amylase from Bacillus lehensis G1

Nor Hasmaliana Abdul Manas, Mohd Anuar Jonet, Abdul Munir Abd. Murad, Nor Muhammad Mahadi, Rosli Md Illias

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Malto-oligosaccharide synthesis using maltogenic amylase often struggles with product re-hydrolyzation. The malto-oligosaccharide synthesis using a maltogenic amylase (MAG1) from Bacillus lehensis G1 was enhanced using a structure-guided protein engineering approach. Mutations decreased the hydrolysis activity of the enzyme and caused various modulations in its transglycosylation properties. W359F, Y377F and M375I mutations caused a reduction in steric interference, an alteration of subsite occupation and an increase in internal flexibility to accommodate longer donor/acceptor molecules for transglycosylation, resulting in an increase in the transglycosylation to hydrolysis ratio of up to 4.0-fold. The increase in active site hydrophobicity that was caused from the W359F and M375I mutations reduced the concentration of maltotriose required for use as a donor/acceptor for transglycosylation to 100 mM and 50 mM, respectively, compared to the 200 mM needed for wild-type. An improvement of the transglycosylation to hydrolysis ratio by 4.2-fold was also demonstrated in each of the mutants. Interestingly, a reduction of steric interference and hydrolysis suppression was caused by the Y377F mutation and introduced a synergistic effect to produce malto-oligosaccharides with a higher degree of polymerization than wild-type. These findings showed that modification of the active site structure imposed various effects on MAG1 activities during malto-oligosaccharide synthesis.

Original languageEnglish
Pages (from-to)1572-1580
Number of pages9
JournalProcess Biochemistry
Volume50
Issue number10
DOIs
Publication statusPublished - 3 Oct 2015

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glucan 1,4-alpha-maltohydrolase
Protein Engineering
Oligosaccharides
Amylases
Bacilli
Bacillus
Hydrolysis
Modulation
Proteins
Mutation
Catalytic Domain
Hydrophobicity
Hydrophobic and Hydrophilic Interactions
Occupations
Polymerization
Enzymes
Molecules

Keywords

  • Malto-oligosaccharide
  • Maltogenic amylase
  • Protein engineering
  • Site-directed mutagenesis
  • Transglycosylation

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Modulation of transglycosylation and improved malto-oligosaccharide synthesis by protein engineering of maltogenic amylase from Bacillus lehensis G1. / Abdul Manas, Nor Hasmaliana; Jonet, Mohd Anuar; Abd. Murad, Abdul Munir; Mahadi, Nor Muhammad; Illias, Rosli Md.

In: Process Biochemistry, Vol. 50, No. 10, 03.10.2015, p. 1572-1580.

Research output: Contribution to journalArticle

Abdul Manas, Nor Hasmaliana ; Jonet, Mohd Anuar ; Abd. Murad, Abdul Munir ; Mahadi, Nor Muhammad ; Illias, Rosli Md. / Modulation of transglycosylation and improved malto-oligosaccharide synthesis by protein engineering of maltogenic amylase from Bacillus lehensis G1. In: Process Biochemistry. 2015 ; Vol. 50, No. 10. pp. 1572-1580.
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