Functionality improvement of fungal lignin peroxidase by DNA shuffling for 2,4-dichlorophenol degradability and H2O2 stability

Title
Functionality improvement of fungal lignin peroxidase by DNA shuffling for 2,4-dichlorophenol degradability and H2O2 stability
Author
이은규
Keywords
Liginin peroxidase; Directed evolution; DNA shuffling; 2,4-DCP degradability; H2O2 stability; Yeast surface display
Issue Date
2008-01
Publisher
ELSEVIER SCIENCE BV
Citation
JOURNAL OF BIOTECHNOLOGY, v. 133, No. 1, Page. 110-115
Abstract
One of the major problems of wild-type lignin peroxidase (LiP) is its inactivity at the presence of excess H2O2 and high concentration of aromatic compounds. Little is known about the substrate-binding site of LiP, and functionality improvement of LiP was not actively tried by genetic engineering and directed evolution. In order to improve LiPs functionality, we performed directed evolution with a colorimetric screening method. Finally, three types of LiP mutants were screened. The catalytic efficiency of the variants toward 2,4-dichlorophenol (DCP) degradation activity and the stability against H2O2 was increased over the wild type. The Km value of the variants toward H2O2 was increased, but Km value toward 2,4-DCP degradation was reduced. Overall, The Kcat/Km values of the mutants toward 2,4-DCP was increased ca. 4-fold, and that toward H2O2 was increased ca. 89-fold. Amino acid sequence analysis indicated that the most of the mutations were located on the enzyme surface. We expect that these results coupled with recombining mutation can be successfully applied to the molecular evolution cycles for screening of LiPs and other oxidative enzymes with improved functionality and stability.
URI
https://www.sciencedirect.com/science/article/pii/S0168165607015726http://repository.hanyang.ac.kr/handle/20.500.11754/76703
ISSN
0168-1656
DOI
10.1016/j.jbiotec.2007.09.008
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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