Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Modulation of Hydrolysis and Transglycosylation Activity of Thermus Maltogenic Amylase by Combinatorial Saturation Mutagenesis

  • Oh, Su-Won (Department of Food Science and Technology, Chungbuk National University) ;
  • Jang, Myoung-Uoon (Department of Food Science and Technology, Chungbuk National University) ;
  • Jeong, Chang-Ku (Department of Food Science and Technology, Chungbuk National University) ;
  • Kang, Hye-Jeong (Department of Food Science and Technology, Chungbuk National University) ;
  • Park, Jung-Mi (Department of Food Science and Technology, Chungbuk National University) ;
  • Kim, Tae-Jip (Department of Food Science and Technology, Chungbuk National University)
  • Published : 2008.08.31
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Abstract

The roles of conserved amino acid residues (Va1329-Ala330-Asn331-Glu332), constituting an extra sugar-binding space (ESBS) of Thermus maltogenic amylase (ThMA), were investigated by combinatorial saturation mutagenesis. Various ThMA mutants were firstly screened on the basis of starch hydrolyzing activity and their enzymatic properties were characterized in detail. Most of the ThMA variants showed remarkable decreases in their hydrolyzing activity, but their specificity against various substrates could be altered by mutagenesis. Unexpectedly, mutant H-16 (Gly-Leu-Val-Tyr) showed almost identical hydrolyzing and transglycosylation activities to wild type, whereas K-33 (Ser-Gly-Asp-Glu) showed an extremely low transglycosylation activity. Interestingly, K-33 produced glucose, maltose, and acarviosine from acarbose, whereas ThMA hydrolyzed acarbose to only glucose and acarviosine-glucose. These results propose that the substrate specificity, hydrolysis pattern, and transglycosylation activity of ThMA can be modulated by combinatorial mutations near the ESBS.

Keywords

References

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