Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions
In previous studies of Lactococcus lactis, the levels of proteins secreted using heterologous signal peptides were observed to be lower than those obtained using the signal peptide from Usp45, the major secreted lactococcal protein. In this study, G1 (the native signal peptide of CGTase) and the sig...
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| Format: | Article |
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Elsevier B.V.
2019
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| _version_ | 1796864787161808896 |
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| author | Mahmud, Hafizah Ismail, Abbas Abdul Rahim, Raha Low, Kheng Oon Md. Illias, Rosli |
| author_facet | Mahmud, Hafizah Ismail, Abbas Abdul Rahim, Raha Low, Kheng Oon Md. Illias, Rosli |
| author_sort | Mahmud, Hafizah |
| collection | ePrints |
| description | In previous studies of Lactococcus lactis, the levels of proteins secreted using heterologous signal peptides were observed to be lower than those obtained using the signal peptide from Usp45, the major secreted lactococcal protein. In this study, G1 (the native signal peptide of CGTase) and the signal peptide M5 (mutant of the G1 signal peptide) were introduced into L. lactis to investigate the effect of signal peptides on lactococcal protein secretion to improve secretion efficiency. The effectiveness of these signal peptides were compared to the Usp45 signal peptide. The highest secretion levels were obtained using the G1 signal peptide. Sequence analysis of signal peptide amino acids revealed that a basic N-terminal signal peptide is not absolutely required for efficient protein export in L. lactis. Moreover, the introduction of a helix-breaking residue in the H-region of the M5 signal peptide caused a reduction in the signal peptide hydrophobicity and decreased protein secretion. In addition, the optimization of cultivation conditions for recombinant G1-CGTase production via response surface methodology (RSM) showed that CGTase activity increased approximately 2.92-fold from 5.01 to 16.89 U/ml compared to the unoptimized conditions. |
| first_indexed | 2024-03-05T20:47:04Z |
| format | Article |
| id | utm.eprints-89226 |
| institution | Universiti Teknologi Malaysia - ePrints |
| last_indexed | 2024-03-05T20:47:04Z |
| publishDate | 2019 |
| publisher | Elsevier B.V. |
| record_format | dspace |
| spelling | utm.eprints-892262021-02-22T06:07:48Z http://eprints.utm.my/89226/ Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions Mahmud, Hafizah Ismail, Abbas Abdul Rahim, Raha Low, Kheng Oon Md. Illias, Rosli TP Chemical technology In previous studies of Lactococcus lactis, the levels of proteins secreted using heterologous signal peptides were observed to be lower than those obtained using the signal peptide from Usp45, the major secreted lactococcal protein. In this study, G1 (the native signal peptide of CGTase) and the signal peptide M5 (mutant of the G1 signal peptide) were introduced into L. lactis to investigate the effect of signal peptides on lactococcal protein secretion to improve secretion efficiency. The effectiveness of these signal peptides were compared to the Usp45 signal peptide. The highest secretion levels were obtained using the G1 signal peptide. Sequence analysis of signal peptide amino acids revealed that a basic N-terminal signal peptide is not absolutely required for efficient protein export in L. lactis. Moreover, the introduction of a helix-breaking residue in the H-region of the M5 signal peptide caused a reduction in the signal peptide hydrophobicity and decreased protein secretion. In addition, the optimization of cultivation conditions for recombinant G1-CGTase production via response surface methodology (RSM) showed that CGTase activity increased approximately 2.92-fold from 5.01 to 16.89 U/ml compared to the unoptimized conditions. Elsevier B.V. 2019-04-20 Article PeerReviewed Mahmud, Hafizah and Ismail, Abbas and Abdul Rahim, Raha and Low, Kheng Oon and Md. Illias, Rosli (2019) Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions. Journal of Biotechnology, 296 . pp. 22-31. ISSN 0168-1656 http://dx.doi.org/10.1016/j.jbiotec.2019.02.013 DOI:10.1016/j.jbiotec.2019.02.013 |
| spellingShingle | TP Chemical technology Mahmud, Hafizah Ismail, Abbas Abdul Rahim, Raha Low, Kheng Oon Md. Illias, Rosli Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| title | Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| title_full | Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| title_fullStr | Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| title_full_unstemmed | Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| title_short | Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| title_sort | enhanced secretion of cyclodextrin glucanotransferase cgtase by lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions |
| topic | TP Chemical technology |
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