Analysis of promoter sequences from Lactobacillus helveticus CNRZ32 and their activity in other lactic acid bacteria

被引:11
作者
Chen, YS
Steele, JL
机构
[1] Mississippi State Univ, Dept Food Sci & Technol, Mississippi State, MS 39762 USA
[2] Univ Wisconsin, Dept Food Sci, Madison, WI 53706 USA
关键词
Escherichia coli; expression; food-grade; Lactobacillus; Lactococcus; promoter;
D O I
10.1111/j.1365-2672.2004.02433.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Aims: To clone and analyse seven putative promoter fragments (pepC, pepN, pepX, pepO, pepE, pepO2, hsp17) from Lactobacillus helveticus CNRZ32 for their expression in Lact. helveticus CNRZ32, Lact. casei ATCC334 and Lactococcus lactis MG1363. Methods and Results: Promoter fragments were fused to the promoter-less beta-glucuronidase (gusA) gene on pNZ272(RBS-ATG-). The resulting constructs were evaluated for their ability to drive the expression of active GusA with 0.5 mmol l(-1) 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide. All promoters except P-pepN::gusA were active in the examined strains. Northern hybridization was performed to examine the promoter strength. Sequence analysis of these promoters identified well conserved putative ribosomal binding and putative -10 hexamers sites. Conclusions: Seven promoter fragments from Lact. helveticus CNRZ32 were recognized in the lactic acid bacteria, Lact. casei ATCC334 and L. lactis MG1363, as well as in Escherichia coli. P-pepN::gusA could not be maintained in the strains examined because of toxicity associated with heterologous protein over-expression driven by P-pepN. Significance and Impact of the Study: This study revealed that desirable levels of heterologous food-grade protein production in GRAS organisms can be obtained with the application of natural promoter fragments from closely related organisms.
引用
收藏
页码:64 / 72
页数:9
相关论文
共 39 条
[1]   TRANSFORMATION OF LACTOBACILLUS-REUTERI WITH ELECTROPORATION - STUDIES ON THE ERYTHROMYCIN RESISTANCE PLASMID PLUL631 [J].
AHRNE, S ;
MOLIN, G ;
AXELSSON, L .
CURRENT MICROBIOLOGY, 1992, 24 (04) :199-205
[2]  
BARTELS HJ, 1987, MILCHWISSENSCHAFT, V42, P83
[3]  
BARTELS HJ, 1987, MILCHWISSENSCHAFT, V42, P139
[4]   Inducible promoter-repressor system from the Lactobacillus casei phage φFSW [J].
Binishofer, B ;
Moll, I ;
Henrich, B ;
Bläsi, U .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (08) :4132-4135
[5]  
Chen YS, 1998, APPL ENVIRON MICROB, V64, P3411
[6]   Identification and characterization of Lactobacillus helveticus PepO2, an endopeptidase with post-proline specificity [J].
Chen, YS ;
Christensen, JE ;
Broadbent, JR ;
Steele, JL .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2003, 69 (02) :1276-1282
[7]   Peptidases and amino acid catabolism in lactic acid bacteria [J].
Christensen, JE ;
Dudley, EG ;
Pederson, JA ;
Steele, JL .
ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY, 1999, 76 (1-4) :217-246
[8]   SEQUENCE-ANALYSIS, DISTRIBUTION AND EXPRESSION OF AN AMINOPEPTIDASE N-ENCODING GENE FROM LACTOBACILLUS-HELVETICUS CNRZ32 [J].
CHRISTENSEN, JE ;
LIN, DL ;
PALVA, A ;
STEELE, JL .
GENE, 1995, 155 (01) :89-93
[9]   CLONING OF PROMOTER-LIKE SEQUENCES FROM LACTOBACILLUS-PARACASEI SUBSP PARACASEI CG11 AND THEIR EXPRESSION IN ESCHERICHIA-COLI, LACTOCOCCUS-LACTIS, AND LACTOBACILLUS-REUTERI [J].
DJORDJEVIC, G ;
BOJOVIC, B ;
BANINA, A ;
TOPISIROVIC, L .
CANADIAN JOURNAL OF MICROBIOLOGY, 1994, 40 (12) :1043-1050
[10]  
ELSODA MA, 1993, FEMS MICROBIOL REV, V12, P239, DOI [10.1111/j.1574-6976.1993.tb00021.x, 10.1016/0168-6445(93)90066-I]