Cloning, Expression, and Chromosomal Stabilization of the Propionibacterium shermanii Proline Iminopeptidase Gene (pip) for Food-Grade Application in Lactococcus lactis
| Autor: | Johan Boot, Marjolein Toonen, Gerard Venema, Inge Deutz, Albert Bolhuis, Aat M. Ledeboer, Kees Leenhouts, Jan Kok |
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| Přispěvatelé: | Groningen Biomolecular Sciences and Biotechnology |
| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
Operon
Lactococcus VECTOR Genetics and Molecular Biology PHAGE medicine.disease_cause Applied Microbiology and Biotechnology Gene dosage Aminopeptidases SUBSP CREMORIS Microbiology ACID BACTERIA Plasmid medicine Escherichia coli AMINOPEPTIDASE GENE Cloning Molecular Promoter Regions Genetic Gene MOLECULAR-CLONING Ecology biology Propionibacterium freudenreichii Lactococcus lactis Propionibacterium SEQUENCE-ANALYSIS Chromosome Mapping DNA Chromosomes Bacterial biology.organism_classification BACILLUS-COAGULANS Biochemistry Food Microbiology SYSTEM Food Science Biotechnology Plasmids |
| Zdroj: | Applied Environmental Microbiology, 64(12), 4736-4742. AMER SOC MICROBIOLOGY |
| ISSN: | 0099-2240 |
| Popis: | Proline iminopeptidase produced by Propionibacterium shermanii plays an essential role in the flavor development of Swiss-type cheeses. The enzyme (Pip) was purified and characterized, and the gene ( pip ) was cloned and expressed in Escherichia coli and Lactococcus lactis , the latter species being an extensively studied, primary cheese starter culture that is less fastidious in its growth condition requirements than P. shermanii . The levels of expression of the pip gene could be enhanced with a factor 3 to 5 by using a strong constitutive promoter in L. lactis or the inducible tac promoter in E. coli . Stable replication of the rolling-circle replicating (rcr) plasmid, used to express pip in L. lactis , could only be obtained by providing the repA gene in trans . Upon the integration of pip , clear gene dosage effects were observed and stable multicopy integrants could be maintained upon growth under the selective pressure of sucrose. The multicopy integrants demonstrated a high degree of stability in the presence of glucose. This study examines the possibilities to overexpress genes that play an important role in food fermentation processes and shows a variety of options to obtain stable food-grade expression of such genes in L. lactis. |
| Databáze: | OpenAIRE |
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