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

Abstract

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.

A review on Lactococcus lactis: from food to factory

“…Prior to this, only prokaryotic membrane protein expressions were performed in L. lactis , mostly with homologous proteins, some which were able to reach up to 30% of total membrane proteins [144]. Most recently, L. lactis was successfully used to express rat and human membrane proteins involved in liver detoxification with higher yields than conventional E. coli and Saccharomyces cerevisiae expression systems [146]. …”

Alterations in Faecal Metagenomics and Serum Metabolomics Indicate Management Strategies for Patients With Budd-Chiari Syndrome

“…Microbiota dysbiosis partially improved following invasive treatment ( Aron-Wisnewsky et al., 2019 ). Lactococcus has been used to prevent and treat various liver injuries and diseases ( Bakari et al., 2016 ; Xu et al., 2019 ). In this study, the population of predominant probiotics such as Lactococcus significantly increased following treatment.…”