EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI
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 i...
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| Format: | Article |
| Language: | English |
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Kemerovo State University
2013-12-01
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| Series: | Foods and Raw Materials |
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| Online Access: | http://jfrm.ru/?page=archive&jrn=1&article=7 |
| _version_ | 1818548478046896128 |
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| author | Babich O.O. Dyshlyuk L. Milent`eva I.S. |
| author_facet | Babich O.O. Dyshlyuk L. Milent`eva I.S. |
| author_sort | Babich O.O. |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-12T08:20:28Z |
| format | Article |
| id | doaj.art-e984b72520214f17aefa9f63a999d766 |
| institution | Directory Open Access Journal |
| issn | 2308-4057 2310-9599 |
| language | English |
| last_indexed | 2024-12-12T08:20:28Z |
| publishDate | 2013-12-01 |
| publisher | Kemerovo State University |
| record_format | Article |
| series | Foods and Raw Materials |
| spelling | doaj.art-e984b72520214f17aefa9f63a999d7662022-12-22T00:31:25ZengKemerovo State UniversityFoods and Raw Materials2308-40572310-95992013-12-0111485310.12737/1542EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLIBabich O.O.0Dyshlyuk L.1Milent`eva I.S. 2Kemerovo Institute of Food Science and TechnologyKemerovo Institute of Food Science and TechnologyKemerovo Institute of Food Science and TechnologyThe 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.http://jfrm.ru/?page=archive&jrn=1&article=7L-phenylalanine ammonia-lyasecloningexpressionrecombinant proteininductionL-phenylalaninephenylketonuria |
| spellingShingle | Babich O.O. Dyshlyuk L. Milent`eva I.S. EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI Foods and Raw Materials L-phenylalanine ammonia-lyase cloning expression recombinant protein induction L-phenylalanine phenylketonuria |
| title | EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI |
| title_full | EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI |
| title_fullStr | EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI |
| title_full_unstemmed | EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI |
| title_short | EXPRESSION OF RECOMBINANT L-PHENYLALANINE AMMONIA-LYASE IN ESCHERICHIA COLI |
| title_sort | expression of recombinant l phenylalanine ammonia lyase in escherichia coli |
| topic | L-phenylalanine ammonia-lyase cloning expression recombinant protein induction L-phenylalanine phenylketonuria |
| url | http://jfrm.ru/?page=archive&jrn=1&article=7 |
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