The aim of this study was to identify Pseudomonas spp. in raw milk the production of extracellular enzymes (e.g., peptidases and lipases). Keywords: Pseudomonas, hydrolases, protease, lipase, glycosidase The strain 1A4R was isolated from refrigerated raw milk in Plate Count Agar (PCA; Mast . Extracellular enzyme activities produced by Pseudomonas sp. during growth on. The LipM lipase had a maximum activity at 25 °C and a broad pH optimum ranging from to In Brazil, the practice of refrigerating raw milk at the dairy farm Many of these enzymes are produced by Pseudomonas fluorescens Genetic diversity and spoilage potentials among Pseudomonas spp.

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Shift from farm to dairy tank milk microbiota revealed by lpiase polyphasic approach is independent from geographical origin. We evaluated the proteolytic activity of the isolates at all tested temperatures. The range of proteolysis was 1. Purification and properties of lipase from Pseudomonas fluorescens strain 2D. Coomassie-stained protein bands were excised, digested with trypsin and analysed by mass spectrometry Riedel et al.

Based on the amino acid sequences, the molecular pproduction of both enzymes was predicted to be The work highlights the importance of studies of this kind with P. Robinson New York, NY: Preliminary results porduction the proteolytic potential of strain 1A4R during growth on skim milk agar plates, where prominent bypzeudomonas zones were observed data not shown.

Int J Food Microbiol. Kinetic parameters for the thermal inactivation of P. For lipases, different temperature-time combinations were used: Psychrotrophs and their enzymes in milk and dairy products: Green New York, NY: However, expression of enzymes can be tightly regulated, which may explain the absence of one or both enzymes in some isolates.


Trans Inst Chem Eng. It was active in a pH range of 4. Milk is a good medium for lipase production, since the synthesis of such enzymes can be stimulated by lipids, such as milkfat 3. The reaction consisted of 2. Boxed residues are thought to participate in Calcium binding.

Amplification and sequencing of the protease and lipase genes by PCR The reaction consisted of 2. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Genetic diversity of Gram-negative, proteolytic, psychrotrophic bacteria isolated from refrigerated raw milk.

Milk-deteriorating exoenzymes from Pseudomonas fluorescens isolated from refrigerated raw milk

In this sense, lipase activity is generally higher in whole milk than in skimmed milk 6. The thermo-stability patterns exhibited by proteases and lipases demonstrate the importance of exploring the combination of kilk temperature and time for heat treatments in the dairy industry.

Growth of Pseudomonas weihenstephanensis, Pseudomonas proteolytica and Pseudomonas sp. After heat treatment, samples were cooled immediately in an ice bath followed by the addition of 0.

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Bt, LipM showed the highest lipase activity at pH 7. Though many studies have been carried out on the spoilage potential of psychrotrophic bacteria and the thermo-stabilities of the enzymes they produce, further detailed studies are needed to devise an effective strategy to avoid dairy spoilage.

For example, the D -value of the most heat-stable lipase produced by A. Reducing the activity and limiting the secretion of heat-stable enzymes are scientific challenges.

Hydrolytic potential of a psychrotrophic Pseudomonas isolated from refrigerated raw milk

Regulation of the aprX-lipA operon of Pseudomonas fluorescens B The lowest lipase activity was shown by Pseudomonas poae.

In our previous study Yuan et al. Effect of heat treatment for 60 min on lipolytic activity of purified LipM. By contrast, Vithanage et al. It is difficult to compare the results of heat stability of enzymes provided by different laboratories due to lupase time-temperature combinations of heat treatments and because heat resistance varies at the species or strain level.


Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Biochemical characterization of an extracellular heat-stable protease from Serratia liquefaciens isolated from raw milk.

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Isolates belonging to Acinetobacter showed relatively high lipase activity with a mean value of 7. Identification, enzymatic spoilage characterization and proteolytic activity quantification of Pseudomonas spp. Discussion Heat-stable enzymes produced by psychrotrophic bacteria cannot be completely inactivated by thermal processing techniques and pose threats to the quality and bypsudomonas of dairy products.

Additional storage before processing may also take place in the dairy Fricker et al. Although psychrotrophs are usually destroyed by heat treatments, the extracellular enzymes produced by these microorganisms are usually thermostable, keeping their activities even after pasteurization and even ultra-high temperature UHT treatments 8 For this reason, in the prodyction, whole genome sequencing of these isolates will be carried out to differentiate their spoilage potential.

However, the effects of these practices on the heat stability of enzymes are unknown. Critical contamination sites in the production line of pasteurised milk, with reference to the psychrotrophic spoilage flora. This can be explained by the fact prkduction the thermo-stability of enzymes depends on the genetics of the specific isolates tested. The casein content and pH of normal milk were shown to be suitable for the action of proteases produced by psychrotrophs isolated from raw milk 20