Characterization of a chimeric enzyme comprising feruloyl esterase and family 42 carbohydrate-binding module |
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Authors: | Takuya Koseki Keiji Mochizuki Hiroe Kisara Akimasa Miyanaga Shinya Fushinobu Tetsuya Murayama Yoshihito Shiono |
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Institution: | (1) Department of Bioresource Engineering, Faculty of Agriculture, Yamagata University, 1-23 Wakaba-machi, Tsuruoka 9978555, Japan;(2) Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 1138657, Japan |
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Abstract: | We engineered a chimeric enzyme (AwFaeA-CBM42) comprising of type-A feruloyl esterase from Aspergillus awamori (AwFaeA) and family 42 carbohydrate-binding module (AkCBM42) from glycoside hydrolase family 54 α-l-arabinofuranosidase of Aspergillus kawachii. The chimeric enzyme was successfully produced in Pichia pastoris and accumulated in the culture broth. The purified chimeric enzyme had an apparent relative molecular mass (M
r) of 53,000. The chimeric enzyme binds to arabinoxylan; this indicates that the AkCBM42 in AwFaeA-CBM42 binds to arabinofuranose
side chain moiety of arabinoxylan. The thermostability of the chimeric enzyme was greater than that of AwFaeA. No significant
difference of the specific activity toward methyl ferulate was observed between the AwFaeA and chimeric enzyme, but the release
of ferulic acid from insoluble arabinoxylan by the chimeric enzyme was approximately 4-fold higher than that achieved by AwFaeA
alone. In addition, the chimeric enzyme and xylanase acted synergistically for the degradation of arabinoxylan. In conclusion,
the findings of our study demonstrated that the components of the AwFaeA-CBM42 chimeric enzyme act synergistically to bring
about the degradation of complex substrates and that the family 42 carbohydrate-binding module has potential for application
in the degradation of polysaccharides. |
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