Аффилиация
a ГОУ ВПО «Кемеровский технологический институт пищевой промышленности»
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Аннотация
Gene , coding L-phenylalanine ammonia-lyase (GenBank: X12702.1) which se-quence was optimized, was cloned into the structure of expressing vector pET28a. Optimization of the expression car-ried out by three parameters (inductor type, induction time and induction temperature) resulted in obtaining a strain-producer of recombinant protein phenylalanine ammonia-lyase with the maximum efficiency of 35±1 % from the total cellular protein when used as an inductor of 0,2 % of lactose (according to Shtudieru), induction time of 18 hours and cultivation temperature of 37 °С. As a result of determination of L-phenylalanine ammonia-lyase solubility it has been shown that recombinant protein is in insoluble fraction by 99 %. The use of 1 mM of IPTG as an inductor instead of 0,2 % of lactose, hasn't changed protein solubility. Protein solubility hasn't been changed as well when cultivating bac-teria at various temperatures: 25 and 37 °С.
Ключевые слова
L-фенилаланин-аммоний-лиаза,
клонирование,
экспрессия,
рекомбинантный белок,
индукция,
L-фенилаланин,
фенилкетонурия,
L-phenylalanine ammonia-lyase,
cloning,
expression,
recombinant protein,
induction,
L-phenylalanine,
phenylketonuria
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