Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it>
<p>Abstract</p> <p>Background</p> <p>High cell density cultures of <it>Pichia pastoris </it>grown on methanol tend to develop yellow colored supernatants, attributed to the release of free flavins. The potential of <it>P. pastoris </it>for flavin...
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| Format: | Article |
| Language: | English |
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BMC
2008-07-01
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| Series: | Microbial Cell Factories |
| Online Access: | http://www.microbialcellfactories.com/content/7/1/23 |
| _version_ | 1831598610627690496 |
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| author | Mattanovich Diethard Marx Hans Sauer Michael |
| author_facet | Mattanovich Diethard Marx Hans Sauer Michael |
| author_sort | Mattanovich Diethard |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>High cell density cultures of <it>Pichia pastoris </it>grown on methanol tend to develop yellow colored supernatants, attributed to the release of free flavins. The potential of <it>P. pastoris </it>for flavin overproduction is therefore given, but not pronounced when the yeast is grown on glucose. The aim of this study is to characterize the relative regulatory impact of each riboflavin synthesis gene. Deeper insight into pathway control and the potential of deregulation is established by overexpression of the single genes as well as a combined deregulation of up to all six riboflavin synthesis genes.</p> <p>Results</p> <p>Overexpression of the first gene of the riboflavin biosynthetic pathway (<it>RIB1</it>) is already sufficient to obtain yellow colonies and the accumulation of riboflavin in the supernatant of shake flask cultures growing on glucose. Sequential deregulation of all the genes, by exchange of their native promoter with the strong and constitutive glyceraldehyde-3-phosphate dehydrogenase promoter (P<sub><it>GAP</it></sub>) increases the riboflavin accumulation significantly.</p> <p>Conclusion</p> <p>The regulation of the pathway is distributed over more than one gene. High cell density cultivations of a <it>P. pastoris </it>strain overexpressing all six <it>RIB </it>genes allow the accumulation of 175 mg/L riboflavin in the supernatant. The basis for rational engineering of riboflavin production in <it>P. pastoris </it>has thus been established.</p> |
| first_indexed | 2024-12-18T14:08:39Z |
| format | Article |
| id | doaj.art-0c4e196ab5d247199a9a2023224c2ae7 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-18T14:08:39Z |
| publishDate | 2008-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-0c4e196ab5d247199a9a2023224c2ae72022-12-21T21:05:11ZengBMCMicrobial Cell Factories1475-28592008-07-01712310.1186/1475-2859-7-23Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it>Mattanovich DiethardMarx HansSauer Michael<p>Abstract</p> <p>Background</p> <p>High cell density cultures of <it>Pichia pastoris </it>grown on methanol tend to develop yellow colored supernatants, attributed to the release of free flavins. The potential of <it>P. pastoris </it>for flavin overproduction is therefore given, but not pronounced when the yeast is grown on glucose. The aim of this study is to characterize the relative regulatory impact of each riboflavin synthesis gene. Deeper insight into pathway control and the potential of deregulation is established by overexpression of the single genes as well as a combined deregulation of up to all six riboflavin synthesis genes.</p> <p>Results</p> <p>Overexpression of the first gene of the riboflavin biosynthetic pathway (<it>RIB1</it>) is already sufficient to obtain yellow colonies and the accumulation of riboflavin in the supernatant of shake flask cultures growing on glucose. Sequential deregulation of all the genes, by exchange of their native promoter with the strong and constitutive glyceraldehyde-3-phosphate dehydrogenase promoter (P<sub><it>GAP</it></sub>) increases the riboflavin accumulation significantly.</p> <p>Conclusion</p> <p>The regulation of the pathway is distributed over more than one gene. High cell density cultivations of a <it>P. pastoris </it>strain overexpressing all six <it>RIB </it>genes allow the accumulation of 175 mg/L riboflavin in the supernatant. The basis for rational engineering of riboflavin production in <it>P. pastoris </it>has thus been established.</p>http://www.microbialcellfactories.com/content/7/1/23 |
| spellingShingle | Mattanovich Diethard Marx Hans Sauer Michael Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it> Microbial Cell Factories |
| title | Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it> |
| title_full | Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it> |
| title_fullStr | Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it> |
| title_full_unstemmed | Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it> |
| title_short | Overexpression of the riboflavin biosynthetic pathway in <it>Pichia pastoris</it> |
| title_sort | overexpression of the riboflavin biosynthetic pathway in it pichia pastoris it |
| url | http://www.microbialcellfactories.com/content/7/1/23 |
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