A genomic search approach to identify esterases in Propionibacterium freudenreichii involved in the formation of flavour in Emmental cheese

Dherbécourt, Julien; Falentin, Hélène; Canaan, Stéphane; Thierry, Anne

doi:10.1186/1475-2859-7-16

Cited by 29 publications

(17 citation statements)

References 43 publications

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“…Here, an iterative strategy was used by integrating heterogeneous localization predictors of lipoprotein-, Tat-, or Sec-dependent signal peptides and transmembrane domains. We showed that one esterase, PF#279, was predicted to be secreted in the medium, and one putative esterase, PF#774, was predicted to be anchored in the plasma membrane, in agreement with previous suggestions made using SignalP as a unique tool (7). In contrast, unclear results were previously found for another esterase, PF#1509, which was clearly predicted as intracellular in the present study.…”

Section: Discussioncontrasting

confidence: 54%

“…However, since the absence of cell lysis was not strictly demonstrated in the latter studies, the esterase activity observed in culture supernatant may have resulted from the release of intracellular enzyme(s) from lysed cells, as previously stressed by Collins et al (2). Twelve putative esterases have been recently identified from the genome sequence of P. freudenreichii CIRM1 T (7). In the present study, we aimed to identify the most probable esterase(s) involved in lipolysis in Emmental cheese, i.e., the esterases that fulfill the following conditions: (i) to be expressed in P. freudenreichii, (ii) to be surface exposed or secreted, and (iii) to be active on milk fat.…”

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confidence: 89%

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Identification of a Secreted Lipolytic Esterase in Propionibacterium freudenreichii , a Ripening Process Bacterium Involved in Emmental Cheese Lipolysis

Dherbécourt

Falentin

Jardin

et al. 2010

Appl Environ Microbiol

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Lipolysis plays an important role in the formation of cheese flavor. In Emmental cheese, the main part of lipolysis has been associated with the presence of Propionibacterium freudenreichii, a species used as a ripening culture. Our aim was to identify the most probable lipolytic esterase(s) involved in cheese lipolysis by P. freudenreichii. Since cheese lipolysis mainly occurs during P. freudenreichii growth, we hypothesized that P. freudenreichii possesses secreted lipolytic esterase(s). For 12 putative esterase genes previously identified from the genome of P. freudenreichii CIRM1, the level of expression was quantified by real-time reverse transcriptase (RT)-PCR, and the subcellular localization of esterases was predicted in silico. The esterase activity in extracellular and intracellular extracts of P. freudenreichii was characterized by zymography, and the extracellular esterases were identified by mass spectrometry. Finally, the best candidate was overexpressed in the same strain. All of the 12 genes encoding putative esterases were expressed. Esterase PF#279 was predicted to be secreted in the medium, PF#774 to be surface exposed, and the 10 remaining putative esterases to be intracellular. Zymography revealed that esterase activities in culture supernatant differed from the ones detected in intracellular extracts. PF#279 was identified as the sole esterase present in culture supernatant. Transformed P. freudenreichii CIRM1 clones overexpressing PF#279 showed 5 to 8 times more lipolytic activity on milk fat than the wild-type strain. Combining in silico, biochemical, and genetic approaches, we showed that PF#279 is the sole secreted esterase in P. freudenreichii and is active on milk fat. Therefore, it is likely a key component in cheese lipolysis by P. freudenreichii.

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Section: Discussioncontrasting

confidence: 54%

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confidence: 89%

Identification of a Secreted Lipolytic Esterase in Propionibacterium freudenreichii , a Ripening Process Bacterium Involved in Emmental Cheese Lipolysis

Dherbécourt

Falentin

Jardin

et al. 2010

Appl Environ Microbiol

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“…Two esterases, one extracellular and other surface-exposed seem to be involved in lipolysis of milk glycerides [63,64]. Furthermore, ten intracellular esterases were found in the P. freudenreichii genome that could be involved in the synthesis of the volatile esters associated with the fruity flavor of cheese [65].…”

Section: Dairy Starters For Swiss-type Cheeses and Other Productsmentioning

confidence: 99%

Dairy Propionibacteria: Less Conventional Probiotics to Improve the Human and Animal Health

Zárate¹

2012

Probiotic in Animals

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“…Much knowledge has been accumulated about diverging genes and converging evolution (Pichersky et al 2006), flavour genes and enzymes (Dherbecourt et al 2008). The toolbox of molecular biology and the volume of sequence databases is ever-increasing (Matsuta et al 2009).…”

Section: Resultsmentioning

confidence: 99%

Biotechnology of flavours—the next generation

2009

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Volatile organic chemicals (flavours, aromas) are the sensory principles of many consumer products and govern their acceptance and market success. Flavours from microorganisms compete with the traditional agricultural sources. Screening for overproducers, elucidation of metabolic pathways and precursors and application of conventional bioengineering has resulted in a set of more than 100 commercial aroma chemicals derived via biotechnology. Various routes may lead to volatile metabolites: De novo synthesis from elementary biochemical units, degradation of larger substrates such as lipids, and functionalization of immediate flavour precursor molecules. More recently, the field was stimulated by the increasing preference of alienated consumers for products bearing the label "natural", and by the vivid discussion on healthy and "functional" food ingredients. The unmistakable call for sustainable sources and environmentally friendly production is forcing the industry to move towards a greener chemistry. Progress is expected from the toolbox of genetic engineering which is expected to help in identifying metabolic bottlenecks and in creating novel high-yielding strains. Bioengineering, in a complementary way, provides promising technical options, such as improved substrate dosage, gas-phase or two-phase reactions and in situ product recovery.

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A genomic search approach to identify esterases in Propionibacterium freudenreichii involved in the formation of flavour in Emmental cheese

Cited by 29 publications

References 43 publications

Identification of a Secreted Lipolytic Esterase in Propionibacterium freudenreichii , a Ripening Process Bacterium Involved in Emmental Cheese Lipolysis

Identification of a Secreted Lipolytic Esterase in Propionibacterium freudenreichii , a Ripening Process Bacterium Involved in Emmental Cheese Lipolysis

Dairy Propionibacteria: Less Conventional Probiotics to Improve the Human and Animal Health

Biotechnology of flavours—the next generation

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