Controlled production of stable heterologous proteins in Lactococcus lactis.
The use of Lactococcus lactis (the most extensively characterized lactic acid bacterium) as a delivery organism for heterologous proteins is, in some cases, limited by low production levels and poor-quality products due to surface proteolysis. In this study, we combined in one L. lactis strain use of the nisin-inducible promoter P(nisA) and inactivation of the extracellular housekeeping protease HtrA. The ability of the mutant strain, designated htrA-NZ9000, to produce high levels of stable proteins was confirmed by using the staphylococcal nuclease (Nuc) and the following four heterologous proteins fused or not fused to Nuc that were initially unstable in wild-type L. lactis strains: (i) Staphylococcus hyicus lipase, (ii) the bovine rotavirus antigen nonstructural protein 4, (iii) human papillomavirus antigen E7, and (iv) Brucella abortus antigen L7/L12. In all cases, protein degradation was significantly lower in strain htrA-NZ9000, demonstrating the usefulness of this strain for stable heterologous protein production.
Miyoshi
A
A
Unité de Recherches Laitières et de Génétique Appliquée, INRA, Domaine de Vilvert, 78352 Jouy en Josas cedex, France.
Poquet
I
I
Azevedo
V
V
Commissaire
J
J
Bermudez-Humaran
L
L
Domakova
E
E
Le Loir
Y
Y
Oliveira
S C
SC
Gruss
A
A
Langella
P
P
D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D002003:Brucella abortus / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D008836:Micrococcal Nuclease; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D027383:Papillomaviridae / Q000235:genetics; D033903:Periplasmic Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D012401:Rotavirus / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C410917:E7 protein, Human papillomavirus type 33; D006360:Heat-Shock Proteins; D009856:Oncogene Proteins, Viral; D033903:Periplasmic Proteins; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C074002:nuc protein, staphylococcus; D012321:DNA-Directed RNA Polymerases; D004720:Endonucleases; D008836:Micrococcal Nuclease; C061682:DegP protease; D012697:Serine Endopeptidases