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Title of article EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI
Authors

Babich O.O. , Kemerovo Institute of Food Science and Technology , bul`v. Stroitelei 47, Kemerovo, 650056 Russia , olich.43@mail.ru

Dyshlyuk L. , Kemerovo Institute of Food Science and Technology , bul`v. Stroitelei 47, Kemerovo, 650056 Russia

Milent`eva I.S. , Kemerovo Institute of Food Science and Technology , bul`v. Stroitelei 47, Kemerovo, 650056 Russia

Section: BIOTECHNOLOGY
Year 2013 Issue 1 DOI 577.112.387.2
Annotation The pal gene coding for L-phenylalanine ammonia-lyase of Rhodosporidium toruloides (GenBank entry no. X12702.1) with optimized sequence was cloned into an expressing vector pET28a. Three parameters of expression (inductor type, duration, and temperature of induction) were optimized, which resulted in a strain producing recombinant L-phenylalanine ammonia-lyase with the maximal productivity, that is, 35 В± 1% to total cell protein, upon utilization of 0.2% lactose (according to Studier) induction during 18 h at 37В°C. The recombinant L-phenylalanine ammonia-lyase was found to be insoluble by 99%. Solubility of the protein did not improve upon utilization of 1 mM IPTG as an inductor instead of 0.2% lactose, or upon bacterium cultivation at various temperatures, that is 25В°C and 37В°C.
Keywords L-phenylalanine ammonia-lyase, cloning, expression, recombinant protein, induction, L-phenylalanine, phenylketonuria
Artice information Received November 30, -0001
Accepted November 30, -0001
Available online November 30, -0001
Imprint article Babich O.O. EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI / Babich O.O., Dyshlyuk L., Milent`eva I.S. // Food and Raw Materials. - 2013. - №1. - С. 48-53
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