Lactococcus lactis: Recent Developments in Functional Expression of Membrane Proteins

Bakari, Sana; André, François; Seigneurin-Berny, Daphné; Delaforge, Marcel; Rolland, Norbert; Frelet-Barrand, Annie

doi:10.1007/978-1-4939-0662-8_5

Cited by 5 publications

(12 citation statements)

References 101 publications

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“…Lactococcus lactis , a Gram-positive bacterium, emerged at the beginning of the twenty-first century as a good alternative to the functional expression of prokaryotic and eukaryotic MPs [ 7 , 11 , 12 ]. This bacterium grows at 30 °C, with a doubling time of 35–60 min, and grows with a fermentative or respiration type of metabolism [ 13 ].…”

Section: Lactococcus Lactismentioning

confidence: 99%

“…Well-characterized and highly versatile, the NICE system has been widely used for the over-expression and the subsequent functional and structural studies of homologous and heterologous MPs [ 12 ]. Moreover, it has been used for other purposes such as in pharmaceutical, medical, biotechnology, and food-technology applications [ 15 , 16 , 33 ].…”

Section: Lactococcus Lactismentioning

confidence: 99%

“…However, L. lactis plasmids (e.g., pNZ8048 or derivatives) cannot be converted into Gateway destination vectors. Therefore, a strategy for the preservation of the correct reading frame has then been established for the rapid transfer of the cDNA from Gateway entry vectors into L. lactis nisin-inducible vectors [ 12 , 50 ]. This strategy allows for the successful expression of MPs from prokaryotic and eukaryotic origins, including proteins which could not be expressed using traditional cloning [ 7 , 51 ].…”

Section: Lactococcus Lactismentioning

confidence: 99%

“…Up to now, 113 MPs from prokaryotic or eukaryotic origin, with diverse topologies and sizes, have been successfully expressed, including 79 in 2014 (12 and the present, Table 2 , Table 3 and Table 4 ). This system also allows for the expression of MPs in their native oligomeric form (homo or heterodimers) [ 11 , 12 ].…”

Section: Expression Of Membrane Proteins Using the Nice Systemmentioning

confidence: 99%

“…The expression levels obtained for the expression of MPs in L. lactis are generally lower than those obtained for the overexpression of the same MPs in E. coli [ 7 , 12 , 60 ]. In some cases, the expression in L. lactis allowed for a higher expression or the expression of proteins usually produced in inclusion bodies in E. coli .…”

Section: Expression Of Membrane Proteins Using the Nice Systemmentioning

confidence: 99%

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Lactococcus lactis, an Attractive Cell Factory for the Expression of Functional Membrane Proteins

Frelet-Barrand

2022

Biomolecules

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Membrane proteins play key roles in most crucial cellular processes, ranging from cell-to-cell communication to signaling processes. Despite recent improvements, the expression of functionally folded membrane proteins in sufficient amounts for functional and structural characterization remains a challenge. Indeed, it is still difficult to predict whether a protein can be overproduced in a functional state in some expression system(s), though studies of high-throughput screens have been published in recent years. Prokaryotic expression systems present several advantages over eukaryotic ones. Among them, Lactococcus lactis (L. lactis) has emerged in the last two decades as a good alternative expression system to E. coli. The purpose of this chapter is to describe L. lactis and its tightly inducible system, NICE, for the effective expression of membrane proteins from both prokaryotic and eukaryotic origins.

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Section: Lactococcus Lactismentioning

confidence: 99%

Section: Lactococcus Lactismentioning

confidence: 99%

Section: Lactococcus Lactismentioning

confidence: 99%

Section: Expression Of Membrane Proteins Using the Nice Systemmentioning

confidence: 99%

Section: Expression Of Membrane Proteins Using the Nice Systemmentioning

confidence: 99%

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Lactococcus lactis, an Attractive Cell Factory for the Expression of Functional Membrane Proteins

Frelet-Barrand

2022

Biomolecules

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Membrane Protein Production in Lactococcus lactis for Functional Studies

Seigneurin-Berny¹,

King²,

Sautron³

et al. 2016

Methods in Molecular Biology

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Due to their unique properties, expression and study of membrane proteins in heterologous systems remains difficult. Among the bacterial systems available, the Gram-positive lactic bacterium, Lactococcus lactis, traditionally used in food fermentations, is nowadays widely used for large-scale production and functional characterization of bacterial and eukaryotic membrane proteins. The aim of this chapter is to describe the different possibilities for the functional characterization of peripheral or intrinsic membrane proteins expressed in Lactococcus lactis.

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Lactococcus lactis is an Efficient Expression System for Mammalian Membrane Proteins Involved in Liver Detoxification, CYP3A4, and MGST1

et al. 2016

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Despite the great importance of human membrane proteins involved in detoxification mechanisms, their wide use for biochemical approaches is still hampered by several technical difficulties considering eukaryotic protein expression in order to obtain the large amounts of protein required for functional and/or structural studies. Lactococcus lactis has emerged recently as an alternative heterologous expression system to Escherichia coli for proteins that are difficult to express. The aim of this work was to check its ability to express mammalian membrane proteins involved in liver detoxification, i.e., CYP3A4 and two isoforms of MGST1 (rat and human). Genes were cloned using two different strategies, i.e., classical or Gateway-compatible cloning, and we checked the possible influence of two affinity tags (6×-His-tag and Strep-tag II). Interestingly, all proteins could be successfully expressed in L. lactis at higher yields than those previously obtained for these proteins with classical expression systems (E. coli, Saccharomyces cerevisiae) or those of other eukaryotic membrane proteins expressed in L. lactis. In addition, rMGST1 was fairly active after expression in L. lactis. This study highlights L. lactis as an attractive system for efficient expression of mammalian detoxification membrane proteins at levels compatible with further functional and structural studies.

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Lactococcus lactis: Recent Developments in Functional Expression of Membrane Proteins

Cited by 5 publications

References 101 publications

Lactococcus lactis, an Attractive Cell Factory for the Expression of Functional Membrane Proteins

Lactococcus lactis, an Attractive Cell Factory for the Expression of Functional Membrane Proteins

Membrane Protein Production in Lactococcus lactis for Functional Studies

Lactococcus lactis is an Efficient Expression System for Mammalian Membrane Proteins Involved in Liver Detoxification, CYP3A4, and MGST1

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