Characterization of a Lactococcus lactis promoter for heterologous protein production
Constitutively active promoter elements for heterologous protein production in Lactococcus lactis are scarce. Here, the promoter of the PTS-IIC gene cluster from L. lactis NZ3900 is described. This promoter was cloned upstream of an enhanced green fluorescent protein, GFPmut3a, and transformed into...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-03-01
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| Series: | Biotechnology Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X17300061 |
| _version_ | 1819293469142679552 |
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| author | Christian E. Ogaugwu Qiuying Cheng Annabeth Fieck Ivy Hurwitz Ravi Durvasula |
| author_facet | Christian E. Ogaugwu Qiuying Cheng Annabeth Fieck Ivy Hurwitz Ravi Durvasula |
| author_sort | Christian E. Ogaugwu |
| collection | DOAJ |
| description | Constitutively active promoter elements for heterologous protein production in Lactococcus lactis are scarce. Here, the promoter of the PTS-IIC gene cluster from L. lactis NZ3900 is described. This promoter was cloned upstream of an enhanced green fluorescent protein, GFPmut3a, and transformed into L. lactis. Transformants produced up to 13.5 μg of GFPmut3a per milliliter of log phase cells. Addition of cellobiose further increased the production of GFPmut3a by up to two-fold when compared to glucose. Analysis of mutations at two specific positions in the PTS-IIC promoter showed that a ‘T’ to ‘G’ mutation within the −35 element resulted in constitutive expression in glucose, while a ‘C’ at nucleotide 7 in the putative cre site enhanced promoter activity in cellobiose. Finally, this PTS-IIC promoter is capable of mediating protein expression in Bacillus subtilis and Escherichia coli Nissle 1917, suggesting the potential for future biotechnological applications of this element and its derivatives. |
| first_indexed | 2024-12-24T04:10:55Z |
| format | Article |
| id | doaj.art-34feb3f283f24cd0927f66c294ba6288 |
| institution | Directory Open Access Journal |
| issn | 2215-017X |
| language | English |
| last_indexed | 2024-12-24T04:10:55Z |
| publishDate | 2018-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj.art-34feb3f283f24cd0927f66c294ba62882022-12-21T17:16:03ZengElsevierBiotechnology Reports2215-017X2018-03-0117C869210.1016/j.btre.2017.11.010Characterization of a Lactococcus lactis promoter for heterologous protein productionChristian E. Ogaugwu0Qiuying Cheng1Annabeth Fieck2Ivy Hurwitz3Ravi Durvasula4New Mexico VA Health Care System, Albuquerque, NM, United StatesNew Mexico VA Health Care System, Albuquerque, NM, United StatesNew Mexico VA Health Care System, Albuquerque, NM, United StatesNew Mexico VA Health Care System, Albuquerque, NM, United StatesNew Mexico VA Health Care System, Albuquerque, NM, United StatesConstitutively active promoter elements for heterologous protein production in Lactococcus lactis are scarce. Here, the promoter of the PTS-IIC gene cluster from L. lactis NZ3900 is described. This promoter was cloned upstream of an enhanced green fluorescent protein, GFPmut3a, and transformed into L. lactis. Transformants produced up to 13.5 μg of GFPmut3a per milliliter of log phase cells. Addition of cellobiose further increased the production of GFPmut3a by up to two-fold when compared to glucose. Analysis of mutations at two specific positions in the PTS-IIC promoter showed that a ‘T’ to ‘G’ mutation within the −35 element resulted in constitutive expression in glucose, while a ‘C’ at nucleotide 7 in the putative cre site enhanced promoter activity in cellobiose. Finally, this PTS-IIC promoter is capable of mediating protein expression in Bacillus subtilis and Escherichia coli Nissle 1917, suggesting the potential for future biotechnological applications of this element and its derivatives.http://www.sciencedirect.com/science/article/pii/S2215017X17300061Lactococcus lactisHeterologous protein expressionPromoterProbiotics |
| spellingShingle | Christian E. Ogaugwu Qiuying Cheng Annabeth Fieck Ivy Hurwitz Ravi Durvasula Characterization of a Lactococcus lactis promoter for heterologous protein production Biotechnology Reports Lactococcus lactis Heterologous protein expression Promoter Probiotics |
| title | Characterization of a Lactococcus lactis promoter for heterologous protein production |
| title_full | Characterization of a Lactococcus lactis promoter for heterologous protein production |
| title_fullStr | Characterization of a Lactococcus lactis promoter for heterologous protein production |
| title_full_unstemmed | Characterization of a Lactococcus lactis promoter for heterologous protein production |
| title_short | Characterization of a Lactococcus lactis promoter for heterologous protein production |
| title_sort | characterization of a lactococcus lactis promoter for heterologous protein production |
| topic | Lactococcus lactis Heterologous protein expression Promoter Probiotics |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X17300061 |
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