Article
TitleCloning and expression of the polyhydroxyalkanote depolymerase gene from Pseudomonas putida, and characterization of the gene product.
AbstractA polyhydroxyalkanote (PHA) depolymerase gene ( pha Z) was cloned by PCR from Pseudomonas putida and over-expressed in Escherichia coli as inclusion bodies. Nucleotide sequence analysis predicted an 852 bp open reading frame encoding a protein of 283 amino acids with a predicted molecular weight of 31283 Da. The deduced amino acid sequence had at least 80% homology to the PHA depolymerase from other Pseudomonas strains and consisted a conserved lipase box-like sequence (G-X-S(102)-X-G). The inclusion bodies were refolded and biochemically characterized. The depolymerase activity was optimal at 40 degrees C and pH 8.
PubMed ID15604801
AuthorJiang Yaqin, Ye Jiang, Wu Haizhen, Zhang Huizhan
JournalBiotechnology letters. 26(20) 1585-8
Date2004 Oct

Protein
Protein namePolyhydroxyalkanote depolymerase
Amino acid sequenceMPQPYVFRTVELDDQSIRTAVRPGKPHLTPLLIFNGIGANLELVFPFIEALDPDLEVIAF
DVPGVGGSSTPRQPYRFPGLAKLTARMLDYLDYGQVNVIGVSWGGALAQQFAHDYPERCK
KLVLAATAAGAVMVPGKPKVLWMMASPRRYVQPSHVMRIAPLIYGGAFRRDPPLAARHAA
KVRSGGKLGYYWQLFAGLGWTSIHWAAQDQPADSGLAGDDDPLIPLINMRLLAWRIPNAQ
LHIIDDGHLFLIIRAEAVAPIIMKFLEQERQRARMHPRPASGT
CommentQ6PLI2
UniProt IDQ6PLI2
Function
Domain

Experiment
Refolding method ☑ dilution ☐ column:filtration ☐ high pressure
☐ dialysis ☐ column:binding ☐ other method
pH 8
Temperature℃ 25
Validation ☑ activity ☐ solubility ☐ non aggregability
☐ circular dichroism ☐ fluorescence tryptophan ☐ nuclear magnetic resonance
☐ crystallization ☐ structure determination

Link to Monash University REFOLD database
Link /refolddatabase/refoldingrecord/1171/

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