Abstract
Objectives
To clone and characterize a novel bi-functional α-amylase/subtilisin inhibitor (LASI) from the rhizome of Ligusticum chuanxiong, a traditional Chinese medicine.
Results
The LASI showed strong homology with members of the Kunitz trypsin inhibitor family. Its putative amino acid sequence has a 40 % identity with that of the α-amylase/subtilisin inhibitor from rice. LASI gene without signal peptide was expressed in E. coli Rosetta. After purification, the recombinant LASI protein was inhibitory against not only α-amylase from porcine pancreas, Helicoverpa armigera, Spodoptera litura and Plutella xylostella, but also subtilisin A, but not against trypsin or chymotrypsin. In addition, the expression level of LASI in rhizome was higher than that in leaf and LASI expression was enhanced by salt, chilling and drought treatment.
Conclusions
This is the first member of the Kunitz-protease inhibitor family identified in traditional Chinese medicine and it might be involved in the plant defense responses against lepidopterous pests, microorganisms and abiotic stresses.
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Acknowledgments
This work was partially funded by National Natural Science Foundation of China (No. 31371232 and No. 31500276), Science and Technology Program for Public Wellbeing from Science and Technology Bureau of Chengdu City (No. 2015-HM01-00051-SF and No. 2015-HM01-00047-SF), the Fundamental Research Funds for the Central Universities of China (No. 2682016YXZT09 and No. 2682016CX099). National Science and Technology Major Project (No. 2014ZX09304307001-019).
Supporting information
Supplementary Fig. 1—Phylogenetic tree of LASI and other α-amylase/subtilisin inhibitors constructed by neighbor-joining algorithm. The numbers at the nodes indicated the bootstrap values. The NCBI protein database accession numbers were shown in brackets.
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Yu, Jh., Li, Yy., Xiang, M. et al. Molecular cloning and characterization of α-amylase/subtilisin inhibitor from rhizome of Ligusticum chuanxiong . Biotechnol Lett 39, 141–148 (2017). https://doi.org/10.1007/s10529-016-2227-8
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DOI: https://doi.org/10.1007/s10529-016-2227-8