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Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions

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

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Official URL: http://dx.doi.org/10.1016/j.jbiotec.2019.02.013

Abstract

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.

Item Type:Article
Uncontrolled Keywords:lactococcus lactis, secretion, signal peptide
Subjects:T Technology > TP Chemical technology
Divisions:Chemical and Energy Engineering
ID Code:89226
Deposited By: Yanti Mohd Shah
Deposited On:22 Feb 2021 14:07
Last Modified:22 Feb 2021 14:07

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