Document Detail


Molecular characterization of phenylalanine ammonia lyase gene from Cistanche deserticola.
MedLine Citation:
PMID:  21104014     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
We cloned the gene, CdPAL1, from Cistanche deserticola callus using RACE PCR with degenerate primers that were designed based on a multiple sequence alignment of known PAL genes from other plant species. The gene shows high homology to other known PAL genes registered in GenBank. The recombinant protein exhibited Michaelis-Menten kinetics with a K (m) of 0.1013 mM, V (max) of 4.858 μmol min(-1), K (cat) of 3.36 S(-1), and K (cat)/K (m) is 33,168 M(-1 )S(-1). The enzyme had an optimal pH of 8.5 and an activation energy of 38.92 kJ mol(-1) when L: -Phenylalanine was used as a substrate; L: -tyrosine cannot be used as substrate for this protein. The optimal temperature was 55°C, and the thermal stability results showed that, after a treatment at 70°C for 20 min, the protein retained 87% activity, while a treatment at 75°C for 20 min resulted in a loss of over 85% of the enzyme activity. Treatment with heavy metal ions (Hg(2+), Pb(2+), and Zn(2+)) showed remarkable inhibitory effects. Among the intermediates from the lignin (cinnamyl alcohol, cinnamyl aldehyde, coniferyl aldehyde, coniferyl alcohol), phenylpropanoid (cinnamic acid, coumaric acid, caffeic acid, and chlorogenic acid) and phenylethanoid (tyrosol and salidroside) biosynthetic pathways, only cinnamic acid showed strong inhibitory effects against CdPAL1 activity with a K (i) of 8 μM. Competitive inhibitor AIP exhibited potent inhibition with K (i) = 0.056 μM.
Authors:
Gao Sheng Hu; Jing Ming Jia; Yeon Jae Hur; Young Soo Chung; Jai Heon Lee; Dae Jin Yun; Woo Sik Chung; Gi Hwan Yi; Tae Ho Kim; Doh Hoon Kim
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Publication Detail:
Type:  Journal Article     Date:  2010-11-20
Journal Detail:
Title:  Molecular biology reports     Volume:  38     ISSN:  1573-4978     ISO Abbreviation:  Mol. Biol. Rep.     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-06-15     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0403234     Medline TA:  Mol Biol Rep     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  3741-50     Citation Subset:  IM    
Affiliation:
College of Life Science and Natural Resources, Dong-A University, Busan, 604-714, Korea.
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