title: Construction and characterization of three protein-targeting expression system in Lactobacillus casei.
abstract: We previously reported that the β-1,4-Mannanase (manB) gene from Bacillus pumilus functions as a good reporter gene in Lactobacillus casei. Two vectors were constructed. One carries the signal peptide of secretion protein Usp45 (SPUsp45) from Lactococcus lactis (pELSH), and the other carries the full-length S-layer protein, SlpA, from L. acidophilus (pELWH). In this work, another vector, pELSPH, was constructed to include the signal peptide of protein SlpA (SPSlpA), and the capacity of all three vectors to drive expression of the manB gene in L. casei was evaluated. The results showed that SPUsp45 is functionally recognized and processed by the L. casei secretion machinery. The SPUsp45-mediated secretion efficiency was ∼87%, and SPSlpA drove the export of secreted ManB with ∼80% efficiency. SPSlpA secretion was highly efficient, and expressed SlpA was anchored to the cell wall by an unknown secretion mechanism. Full-length SlpA drove the cell wall-anchored expression of an SlpA-ManB fusion protein but at a much lower level than that of protein SlpA.
authors.affiliation: State Key Laboratory of Dairy Biotechnology, Technology Center of Bright Dairy and Food Co., Ltd., 1518 Jiangchang Road (W), Shanghai 200436, China.
authors.forename: Jinzhong
authors.initials: J
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Yexia
authors.initials: Y
authors.lastname: Zou
authors.affiliation: State Key Laboratory of Dairy Biotechnology, Technology Center of Bright Dairy and Food Co., Ltd., 1518 Jiangchang Road (W), Shanghai 200436, China.
authors.forename: Chengjie
authors.initials: C
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Ge
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China Zhengjun.chen@gmail.com.
authors.forename: Zhengjun
authors.initials: Z
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China Department of Biology, University of Copenhagen, Biocenter, Ole Maaloes Vej 5, DK-2200 Copenhagen N, Denmark.
authors.forename: Qunxin
authors.initials: Q
authors.lastname: She
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D017930:Genes, Reporter; D005822:Genetic Vectors; D007780:Lacticaseibacillus casei / Q000235:genetics* / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics; D009097:Multienzyme Complexes / Q000235:genetics*; D009713:Nucleotidyltransferases / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008562:Membrane Glycoproteins; D009097:Multienzyme Complexes; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C055764:S-layer proteins; C070683:surface layer protein A, Bacteria; C093300:ManB protein, bacteria; D009713:Nucleotidyltransferases; D008359:Mannose-6-Phosphate Isomerase
keywords: Lactobacillus casei; S-layer; anchor expression; secretion efficiency; secretion expression; β-1,4-mannanase