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Growth of Pseudomonas weihenstephanensis, Pseudomonas proteolytica and Pseudomonas sp. Impact of residual heat-stable enzyme activity on stability of UHT milk during shelf-life.

One of the reasons for spoilage of UHT milk during shelf-life is the presence of residual proteolytic activity produced from Pseudomonas spp.

The aim of this study was to describe the product defects occurring in indirectly heated UHT milk during shelf-life, and to establish a correlation between proteolytic activity and onset of product spoilage.

Inactivation kinetics of the peptidases were determined. In UHT milk, product defects occurred in the order: A linear correlation was found between proteolytic activity and onset of product defects, apart from onset of gelation.

Thermostability of peptidases secreted by microorganisms associated with raw milk. Peptidases of psychrotolerant microorganisms are known to be thermostable and withstand the thermal processing of milk products.

The protective effect of milk protein and influence of autoproteolysis were shown in extensive studies. The peptidases withstood ultrahigh temperature treatment with residual activities of The valuable lactose derivatives lactulose and epilactose can be derived from lactose either by the Lobry de Bruyn-Alberda van Ekenstein transformation during heat treatments or by enzymatic conversion using cellobiose 2- epimerases EC 5.

The chromatographic determination of lactose, lactulose, and epilactose inmilk is challenging, due to the variable ratio of the three saccharides and their similar retention properties.

In this work, a dual highperformance liquid chromatography HPLC analysis for the quantification of lactose, lactulose, and epilactose in milk samples was developed and validated.

The samples originated from an enzymatic lactose conversion using the cellobiose 2- epimerase from Caldicellulosiruptor saccharolyticus.

Application of this enzyme led to the formation of high lactulose concentrations The dual HPLC analysis utilized a combination of two chromatographic separation techniques, configured in two parallel systems.

After precolumn derivatization, the samples were analyzed as follows: Method 1 determined the concentration of lactose and epilactose using a C18 column with an ion-pair reagent as eluent, coupled with a UV detector.

Both methods were validated in terms of linearity, precision and recovery. The revealing detection limits in the milk samples were 3.

The dual HPLC analysis presented allows accurate lactose, lactulose, and epilactose separation in complex food matrices such as milk.

Analysis of raw cow's milk and semi-finished milk products microbiota yielded seven isolates assigned to the genus Pseudomonas which formed two individual groups in a phylogenetic analysis based on partial rpoD and 16S rRNA gene sequences.

The two groups could be differentiated from each other and also from their closest relatives as well as from the type species Pseudomonas aeruginosa by phenotypic and chemotaxonomic characterisation and ANIb values calculated from draft genome assemblies.

ANIb values within the groups are higher than Funktionelle Proteinhydrolysate - Potenzial von Peptidasen für die Proteinmodifikation in Lebensmitteln.

Enzyme sind aus unserem täglichen Leben nicht mehr wegzudenken. Sie werden in nahezu allen Bereichen, der landwirtschaftlichen-, der pharmazeutischen-, der chemischenoder der lebensmitteltechnologischen Industrie eingesetzt.

Die Vorteile von Enzymen sind vielfältig. Im Gegensatz zu chemischen Prozessen, bei denen gesundheitsgefährdende oder geschmackbeeinflussende Nebenprodukte im Lebensmittel entstehen können, laufen enzymatische Prozesse selektiv bei moderaten Temperaturen und pH-Werten ab.

Ein weiterer Vorteil enzymatischer Prozesse ist, dass die Zusammensetzung des Produktes hier: Hydrolysat je nach Selektivität bzw.

Spezifität des eingesetzten Enzyms hier: Peptidase , gezielt beeinflusst werden kann. Wheat gluten hydrolysis using isolated Flavourzyme peptidases: Product inhibition and determination of synergistic effects using response surface methodology.

However, a biochemical characterization of the Flavourzyme peptidases is difficult, because obtaining purified proteins is essential when functional and structural characterization studies are targeted.

Key enzyme activities three endopeptidases, two aminopeptidases, two dipeptidyl peptidases have recently been identified and isolated from this commercially available enzyme preparation.

The impact and the synergism of theses peptidases on the complex wheat gluten hydrolysis are yet unclear. However, the knowledge about the latter is crucial for an efficient protein hydrolysis.

In the present study, we determined the product inhibition for the seven isolated peptidases and analyzed the impact of each peptidase on the wheat gluten hydrolysis using response surface methodology.

In general, both aminopeptidases and the three endopeptidases were of major importance. One of the endopeptidases alkaline protease 1 was least affected by product inhibition and showed the highest effect on the wheat gluten hydrolysis.

In the case of the aminopeptidases, the leucine aminopeptidase 2 showed a higher impact on the hydrolysis compared to the leucine aminopeptidase A, but exhibited the highest product inhibition sensitivity.

The dipeptidyl peptidases were of only minor impact on the wheat gluten hydrolysis. Enzymatic production of lactulose and epilactose in milk.

The enzymatic production of lactulose was described recently through conversion of lactose by a thermophilic cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus CsCE.

In the current study, we examined the application of CsCE for lactulose and epilactose production in milk 1.

The conversion of milk lactose initial lactose content of This enzymatic lactose conversion resulted in The time courses of lactose conversion by CsCE suggested that first epilactose formed and afterward lactulose via epilactose.

To the best of our knowledge, this is the first time that an enzyme has produced lactulose directly in milk in situ at industrially relevant temperatures.

Published by Elsevier Inc. Test system for direct measurement of lipase activities in milk samples. European Biotechnology Congress Biodiversity of refrigerated raw milk microbiota and their enzymatic spoilage potential.

The refrigerated storage of raw milk selects for psychrotolerant microorganisms, many of which produce peptidases and lipases. Some of these enzymes are heat resistant and are not sufficiently inactivated by pasteurisation or even ultra-high temperature UHT treatment.

In the current study, 20 different raw cow's milk samples from single farms and dairy bulk tanks were analysed close to delivery to the dairies or close to processing in the dairy for their cultivable microbiota as well as the lipolytic and proteolytic potential of the isolated microorganisms.

Altogether, isolates have been identified and assigned to species and 61 genera. The potential of the isolates to produce lipases or peptidases followed in many cases a genus or group specific pattern.

All isolates identified as members of the genus Pseudomonas exhibited mainly lipolytic and proteolytic activity or solely proteolytic activity.

On the other hand, nearly all isolates of the genus Acinetobacter were lipolytic but not proteolytic. Microbial lipases may be produced during milk storage and processing.

This can lead to organoleptic changes in the corresponding dairy products. Thus, monitoring of lipase activity in milk is desirable.

Turbidity of milk prevents a direct photometric measurement of lipase activity using chromophore- or fluorophore-based assays.

Laborious pretreatments or alternative analytical methods normally have to be used. With the method for the determination of lipolytic activity MeDeLi proposed here, it is possible to measure lipase activity directly in the natural milk utilizing tailored fluorometric substrates.

Only a defatting step is carried out initially for the MeDeLi. Then, the conversion of added lipase substrate is carried out in the unmodified milk without addition of any solutions or any enzyme extraction procedure which may influence the enzyme activity.

Thereafter, the milk sample is treated with two solutions to remove the turbidity of milk by dissolution. A valid and sensitive fluorometric measurement is then possible.

The applicability of the MeDeLi was demonstrated in comparison with tests published previously: Raw milk, milk products, and spoiled milk samples were also investigated with the MeDeLi.

Isolation and characterisation of a heat-resistant peptidase from Pseudomonas panacis withstanding general UHT processes. Quantification of the proteolytic and lipolytic activity of microorganisms isolated from raw milk.

Each year we treat thousands of patients covering a wide range of symptoms and conditions including the Menopause, Andropause, Thyroid Dysfunction and PMS.

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Thermostability of peptidases secreted by microorganisms associated with raw milk. Peptidases of psychrotolerant microorganisms are known to be thermostable and withstand the thermal processing of milk products.

The protective effect of milk protein and influence of autoproteolysis were shown in extensive studies. The peptidases withstood ultrahigh temperature treatment with residual activities of The valuable lactose derivatives lactulose and epilactose can be derived from lactose either by the Lobry de Bruyn-Alberda van Ekenstein transformation during heat treatments or by enzymatic conversion using cellobiose 2- epimerases EC 5.

The chromatographic determination of lactose, lactulose, and epilactose inmilk is challenging, due to the variable ratio of the three saccharides and their similar retention properties.

In this work, a dual highperformance liquid chromatography HPLC analysis for the quantification of lactose, lactulose, and epilactose in milk samples was developed and validated.

The samples originated from an enzymatic lactose conversion using the cellobiose 2- epimerase from Caldicellulosiruptor saccharolyticus. Application of this enzyme led to the formation of high lactulose concentrations The dual HPLC analysis utilized a combination of two chromatographic separation techniques, configured in two parallel systems.

After precolumn derivatization, the samples were analyzed as follows: Method 1 determined the concentration of lactose and epilactose using a C18 column with an ion-pair reagent as eluent, coupled with a UV detector.

Both methods were validated in terms of linearity, precision and recovery. The revealing detection limits in the milk samples were 3.

The dual HPLC analysis presented allows accurate lactose, lactulose, and epilactose separation in complex food matrices such as milk.

Analysis of raw cow's milk and semi-finished milk products microbiota yielded seven isolates assigned to the genus Pseudomonas which formed two individual groups in a phylogenetic analysis based on partial rpoD and 16S rRNA gene sequences.

The two groups could be differentiated from each other and also from their closest relatives as well as from the type species Pseudomonas aeruginosa by phenotypic and chemotaxonomic characterisation and ANIb values calculated from draft genome assemblies.

ANIb values within the groups are higher than Funktionelle Proteinhydrolysate - Potenzial von Peptidasen für die Proteinmodifikation in Lebensmitteln.

Enzyme sind aus unserem täglichen Leben nicht mehr wegzudenken. Sie werden in nahezu allen Bereichen, der landwirtschaftlichen-, der pharmazeutischen-, der chemischenoder der lebensmitteltechnologischen Industrie eingesetzt.

Die Vorteile von Enzymen sind vielfältig. Im Gegensatz zu chemischen Prozessen, bei denen gesundheitsgefährdende oder geschmackbeeinflussende Nebenprodukte im Lebensmittel entstehen können, laufen enzymatische Prozesse selektiv bei moderaten Temperaturen und pH-Werten ab.

Ein weiterer Vorteil enzymatischer Prozesse ist, dass die Zusammensetzung des Produktes hier: Hydrolysat je nach Selektivität bzw.

Spezifität des eingesetzten Enzyms hier: Peptidase , gezielt beeinflusst werden kann. Wheat gluten hydrolysis using isolated Flavourzyme peptidases: Product inhibition and determination of synergistic effects using response surface methodology.

However, a biochemical characterization of the Flavourzyme peptidases is difficult, because obtaining purified proteins is essential when functional and structural characterization studies are targeted.

Key enzyme activities three endopeptidases, two aminopeptidases, two dipeptidyl peptidases have recently been identified and isolated from this commercially available enzyme preparation.

The impact and the synergism of theses peptidases on the complex wheat gluten hydrolysis are yet unclear. However, the knowledge about the latter is crucial for an efficient protein hydrolysis.

In the present study, we determined the product inhibition for the seven isolated peptidases and analyzed the impact of each peptidase on the wheat gluten hydrolysis using response surface methodology.

In general, both aminopeptidases and the three endopeptidases were of major importance. One of the endopeptidases alkaline protease 1 was least affected by product inhibition and showed the highest effect on the wheat gluten hydrolysis.

In the case of the aminopeptidases, the leucine aminopeptidase 2 showed a higher impact on the hydrolysis compared to the leucine aminopeptidase A, but exhibited the highest product inhibition sensitivity.

The dipeptidyl peptidases were of only minor impact on the wheat gluten hydrolysis. Enzymatic production of lactulose and epilactose in milk.

The enzymatic production of lactulose was described recently through conversion of lactose by a thermophilic cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus CsCE.

In the current study, we examined the application of CsCE for lactulose and epilactose production in milk 1. The conversion of milk lactose initial lactose content of This enzymatic lactose conversion resulted in The time courses of lactose conversion by CsCE suggested that first epilactose formed and afterward lactulose via epilactose.

To the best of our knowledge, this is the first time that an enzyme has produced lactulose directly in milk in situ at industrially relevant temperatures.

Published by Elsevier Inc. Test system for direct measurement of lipase activities in milk samples. European Biotechnology Congress Biodiversity of refrigerated raw milk microbiota and their enzymatic spoilage potential.

The refrigerated storage of raw milk selects for psychrotolerant microorganisms, many of which produce peptidases and lipases.

Some of these enzymes are heat resistant and are not sufficiently inactivated by pasteurisation or even ultra-high temperature UHT treatment.

In the current study, 20 different raw cow's milk samples from single farms and dairy bulk tanks were analysed close to delivery to the dairies or close to processing in the dairy for their cultivable microbiota as well as the lipolytic and proteolytic potential of the isolated microorganisms.

Altogether, isolates have been identified and assigned to species and 61 genera. The potential of the isolates to produce lipases or peptidases followed in many cases a genus or group specific pattern.

All isolates identified as members of the genus Pseudomonas exhibited mainly lipolytic and proteolytic activity or solely proteolytic activity.

On the other hand, nearly all isolates of the genus Acinetobacter were lipolytic but not proteolytic. Microbial lipases may be produced during milk storage and processing.

This can lead to organoleptic changes in the corresponding dairy products. Thus, monitoring of lipase activity in milk is desirable. Turbidity of milk prevents a direct photometric measurement of lipase activity using chromophore- or fluorophore-based assays.

Laborious pretreatments or alternative analytical methods normally have to be used. With the method for the determination of lipolytic activity MeDeLi proposed here, it is possible to measure lipase activity directly in the natural milk utilizing tailored fluorometric substrates.

Only a defatting step is carried out initially for the MeDeLi. Then, the conversion of added lipase substrate is carried out in the unmodified milk without addition of any solutions or any enzyme extraction procedure which may influence the enzyme activity.

Thereafter, the milk sample is treated with two solutions to remove the turbidity of milk by dissolution. A valid and sensitive fluorometric measurement is then possible.

The applicability of the MeDeLi was demonstrated in comparison with tests published previously: Raw milk, milk products, and spoiled milk samples were also investigated with the MeDeLi.

Isolation and characterisation of a heat-resistant peptidase from Pseudomonas panacis withstanding general UHT processes.

Quantification of the proteolytic and lipolytic activity of microorganisms isolated from raw milk. Extracellular peptidases from insect- and compost-associated microorganisms: Screening for microorganisms with an enzyme activity for a specific application is challenging.

In this study, we showed a successful way to isolate microorganisms from insects and compost samples, which had a desired enzyme activity, namely the hydrolysis of wheat gluten.

This was realized by a two-step screening approach. The first step was an agar plate screening assay for extracellular peptidase activity, and the most promising organisms were further tested in a second screening step: The best isolate, which was identified as a Bacillus subtilis species, was cultivated in a bioreactor 35 L scale to produce a larger amount of the extracellular peptidases.

The peptidase activities in the cell-free culture medium were partially purified, as it is common for technical enzyme preparations for the food industry.

Get in touch with our dedicated patient care team to book your appointment today. Looking to find out more about bio-identical hormone replacement therapy BHRT?

This comprehensive guide will help answer your questions. We treat both women and men at the Marion Gluck Clinic. Find out more about which hormonal conditions we treat and how your symptoms can be managed using tailored bio-identical hormones.

The Home of Bio-identical Hormones. What We Treat For Women. Treatments We help our patients achieve the best possible results through a wide variety of hormonal treatments for both women and men.

The best consultation I have had in a clinic. My doctor was friendly and made me feel very comfortable, and seemed highly competent and experienced.

She was thorough, and took her time during the consultation. I arrived to my appointment and was seen without delay.

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