Engineering of dual-functional hybrid glucanases

Wei Chun Liu, Yu Shiun Lin, Wen Yih Jeng, Je Hsin Chen, Andrew H.J. Wang, Lie Fen Shyur

研究成果: 雜誌貢獻文章同行評審

7 引文 斯高帕斯(Scopus)

摘要

1,3-1,4-β-d-Glucanase (lichenase) and 1,3-β-d-glucanase (laminarinase) are fibrolytic enzymes which play an important role in the hydrolysis of polysaccharide components. Both of these glucanases have been employed in a number of industrial applications. This study aims to improve or combine the novel properties of both glucanases in an attempt to create desirable hybrid enzymes with economic benefits for industrial applications. A truncated and mutated 1,3-1,4-β-d-glucanase gene (TFsW203F) from Fibrobacter succinogenes, and a 1,3-β-d-glucanase gene (TmLam) from hyperthermophilic Thermotoga maritima were used as target enzymes. The substrate-binding domains (TmB1 and TmB2) and the catalytic domain (TmLamCD) of TmLam were ligated to the N- or C-terminus of TFsW203F to create four hybrid enzymes, TmB 1-TFsW203F, TFsW203F-TmB2, TmB 1-TFsW203F-TmB2 and TFsW203F- TmLamCD. The results obtained from kinetic studies show that increased specific activities and turnover rate for lichenan and laminarin were observed in TmB1-TFsW203F-TmB2 and TFs W203F-TmLamCD, respectively. Furthermore, fluorescence and circular dichroism spectrometric analyses indicated that the hybrid TFs W203F-TmLamCD was structurally more stable than the parental TFsW203F, which was attributed to an improved thermal tolerance of the hybrid enzyme. This study has been successful in creating bifunctional hybrid glucanases with dual substrate catalytic functions which warrant further evaluation of their possible use in industrial applications.

原文英語
頁(從 - 到)771-780
頁數10
期刊Protein Engineering, Design and Selection
25
發行號11
DOIs
出版狀態已發佈 - 11月 2012
對外發佈

ASJC Scopus subject areas

  • 生物技術
  • 生物工程
  • 生物化學
  • 分子生物學

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