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Enhancing the Enzymatic Activity of the Multifunctional β-Glycosyl Hydrolase (Cel44C-Man26AP558) from Paenibacillus polymyxa GS01 Using DNA Shuffling
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  • Journal title : The Korean Journal of Microbiology
  • Volume 48, Issue 2,  2012, pp.73-78
  • Publisher : The Microbiological Society of Korea
  • DOI : 10.7845/kjm.2012.48.2.073
 Title & Authors
Enhancing the Enzymatic Activity of the Multifunctional β-Glycosyl Hydrolase (Cel44C-Man26AP558) from Paenibacillus polymyxa GS01 Using DNA Shuffling
Kang, Young-Min; Kang, Tae-Ho; Yun, Han-Dae; Cho, Kye-Man;
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 Abstract
We previously reported that the truncated Cel44C- -glycosyl hydrolase protein exhibits multifunctional activities, including cellulase, xylanase, and lichenase. DNA shuffling of the truncated Cel44C- enzyme was performed to enhance the enzymatic activity of the multifunctional -glycosyl hydrolase. Two mutant enzymes, M2Cel44C- that carries one mutation (P438A) and M21Cel44C- that carries two mutations (A273T and P438A) were obtained. The enzymatic activity of the M21Cel44C- double mutant was lower than enzymatic activity of the single mutant (M2Cel44C-). However, both mutants displayed the enhancements in their enzyme activities that were - to 2.2-fold higher than the original enzymatic activity in Cel44C-. In particular, the mutant M2Cel44C- exhibited an approximate 1.5- to 2.2-fold increase in the cellulase, xylanase, and lichenase activities in comparison with the control (Cel44C-). The optimum cellulase, linchenase, and xylanase activities of -glycosyl hydrolase were observed at pH 7.0, pH 7.0 and pH 6.0, respectively. These results, therefore, suggest that the amino acid residue Ala438 plays important roles in the enhancement of the activity of multifunctional -glycosyl hydrolase.
 Keywords
Paenibacillus polymyxa GS01;Cel44C-;DNA shuffling;multifunctional -glycosyl hydrolase;
 Language
English
 Cited by
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1.
Construction and expression of a novel Paenibacillus polymyxa GS01 bifunctional xyn43A-lin16A gene through overlap extension PCR, Journal of the Korean Society for Applied Biological Chemistry, 2013, 56, 4, 383  crossref(new windwow)
2.
Metagenomics of un-culturable bacteria in cow rumen: Construction of cel9E–xyn10A fusion gene by site-directed mutagenesis, Journal of Molecular Catalysis B: Enzymatic, 2015, 113, 29  crossref(new windwow)
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