Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris
| Autor: | Chieh-Yang Cheng, Lu-Sheng Hsieh, Hung-Chi Pan, Chuan-Shan Yeh, Ping-Du Lee, Chien-Chih Yang |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Ammonia-Lyases
Bambusa Phenylalanine ammonia-lyase Biology Molecular cloning Bambusa oldhamii medicine.disease_cause Pichia law.invention Pichia pastoris Open Reading Frames law Escherichia coli medicine Gene Library Phenylalanine Ammonia-Lyase Phenylpropanoid biology.organism_classification Molecular biology Recombinant Proteins humanities Biochemistry Recombinant DNA Biotechnology Homotetramer |
| Zdroj: | Protein Expression and Purification. 71:224-230 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2010.01.009 |
| Popis: | Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is the first committed enzyme of phenylpropanoid pathway. A PAL gene, designated as BoPAL2, was cloned from a Bambusa oldhamii cDNA library. The open reading frame of BoPAL2 was 2142bp in size encoding a 713-amino acid polypeptide. BoPAL2 was heterologous expressed in Escherichia coli and Pichia pastoris. The recombinant proteins were exhibited PAL and tyrosine ammonia-lyase activities. The recombinant BoPAL2 had a subunit mass of 80kDa and existed as a homotetramer. The optimum temperature and pH of BoPAL2 were 50-60 degrees C and 8.5-9.0, respectively. The K(m) and k(cat) values of BoPAL2 expressed in E. coli were 250microM and 10.12s(-1). The K(m) and k(cat) values of BoPAL2 expressed in P. pastoris were 331microM and 16.04s(-1). The recombinant proteins had similar biochemical properties and kinetic parameters with PALs reported in other plants. |
| Databáze: | OpenAIRE |
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