Determination of proteolytic activity in different milk systems

Bendicho, Sı́lvia; Martí, G.; Hernandez, Thomas; Martín, O.

doi:10.1016/s0308-8146(02)00126-7

Cited by 36 publications

(22 citation statements)

References 14 publications

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“…Protease activity was determined according to Bendicho et al [30]. Enzyme extract (50 lL) was incubated with 500 lL of a 0.5% (m/v) azocasein (Sigma-Aldrich, St Louis, USA) solution, for 5 min.…”

Section: Assays Of Enzyme Activities and Products Of Hydrolysismentioning

confidence: 99%

Valorization of Residual Agroindustrial Cakes by Fungal Production of Multienzyme Complexes and Their Use in Cold Hydrolysis of Raw Starch

Castro

Andréa

Carvalho

et al. 2011

Waste Biomass Valor

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In this work four agroindustrial cakes were used as raw material both for production of enzyme pools containing amylases and accessory hydrolases by solidstate fermentation (SSF) and for cold starch hydrolysis. Eight fungal strains from the genera Aspergillus and Penicillium were screened for enzyme production, and their enzyme extracts were then evaluated in the hydrolysis of raw cakes. Babassu cake was the most suitable raw material for endoamylases, exoamylases and proteases production. The highest activities of these three enzymes were produced by A. awamori IOC-3914 (29.8 U g -1 ), A. wentii (47.8 U g -1 ) and P. verrucosum (27.5 U g -1 ), respectively. Regarding cellulases and xylanases, maximum activities (12.8 and 598.0 U g -1 , respectively) were obtained by A. awamori IOC-3915 in castor seed residue. Saccharification studies showed a flexible applicability of the raw extracts to hydrolyze different cakes. A maximum total reducing sugars concentration of 13.9 g L -1 was obtained from babassu cake using a raw enzyme extract produced by A. awamori IOC-3915, without any concentration or purification steps. The present results demonstrate that a low-cost SSF process can supply enzyme extracts with a high potential for application in the cold hydrolysis of raw starch from agroindustrial cakes.

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Section: Assays Of Enzyme Activities and Products Of Hydrolysismentioning

confidence: 99%

Valorization of Residual Agroindustrial Cakes by Fungal Production of Multienzyme Complexes and Their Use in Cold Hydrolysis of Raw Starch

Castro

Andréa

Carvalho

et al. 2011

Waste Biomass Valor

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“…Samples should be pre-treated to provide absorbances between 0.100 and 1.000 on instrument display, but both used and modified methods even with prolonged reaction time did not offer suitable results (data not shown). The least detection and quantification limits of the method, as published by Bendicho et al (2002), were 2.29 mU/ml and 7.64 mU/ml of bacillus subtilis protease added to milk, respectively, but these activities may be higher than those present in analysed samples.…”

Section: Resultsmentioning

confidence: 82%

“…Method used by Bendicho et al (2002) was modified. 0.2 ml of milk sample was added to 2 ml of 1% azo-casein solution in 0.1 mol/l TRIS.HCl buffer pH 7.5 and incubated at 37°C for 1 hour.…”

Section: Methodsmentioning

confidence: 99%

Problems with detection of proteolytic microorganisms and their undesirable activities in milk

Němečková¹,

Pechacova²,

Roubal³

2009

Czech J. Food Sci.

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Abstract:Occurrence of proteolytic enzymes in milk is often associated with technological problems and sensory, rheological and functional defects of final dairy products. Thus, the simple, cost-effective and available laboratory method for evaluation of undesirable proteolysis risk is needed. In our work we have tested cultivation plate methods and chemical methods (formol titration, ammonium reflectometric determination, the Kjeldahl method, the agarwell diffusion assay and spectrofotometry after cleavage of azo-casein) to choose the proper ones which can provide information on undesirable proteolytic changes especially in raw milk. Although the microbiological analyses cannot detect enzymes indigenous to milk, but only the quantity of producers of microbial enzymes, they seem to be the most acceptable, particularly usage of the Glucose-Trypton-Yeast Extract agar with 10% vol. of sterile milk added before pouring onto plates (incubation at 30°C for 72 h). The chemical methods are not sensitive enough to analyse the real milk samples.

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“…APM activity modulation assays The enzymatic activity of APM was determined by the hydrolysis of azocasein (Sigma Chemical Co., St Louis, MO, USA) at 378C, as it has been previously described (Bendicho et al, 2002). Briefly, APM was incubated in the presence of azocasein (5 mg/ml) in acetate buffer 0.1 M pH 5.0, for 90 min at 378C.…”

Section: Methodsmentioning

confidence: 99%

Activity modulation of microbial enzymes by llama (Lama glama) heavy-chain polyclonal antibodies during in vivo immune responses

Ferrari

Weill

Paz

et al. 2012

Animal

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Since they were first described in 1993, it was found that recombinant variable fragments (rVHHs) of heavy-chain antibodies (HCAbs) from Camelidae have unusual biophysical properties, as well as a special ability to interact with epitopes that are cryptic for conventional Abs. It has been assumed that in vivo raised polyclonal HCAbs (pHCAbs) should behave in a similar manner than rVHHs; however, this assumption has not been tested sufficiently. Furthermore, our own preliminary work on a single serum sample from a llama immunized with a b-lactamase, has suggested that pHCAbs have no special ability to down-modulate catalytic activity. In this work, we further explored the interaction of pHCAbs from four llamas raised against two microbial enzymes and analyzed it within a short and a long immunization plan. The relative contribution of pHCAbs to serum titer was found to be low compared with that of the most abundant conventional subisotype (IgG 1 ), during the whole immunization schedule. Furthermore, pHCAbs not only failed to inhibit the enzymes, but also activated one of them. Altogether, these results suggest that raising high titer inhibitory HCAbs is not a straightforward strategy -neither as a biotechnological strategy nor in the biological context of an immune response against infection -as raising inhibitory rVHHs.

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Determination of proteolytic activity in different milk systems

Cited by 36 publications

References 14 publications

Valorization of Residual Agroindustrial Cakes by Fungal Production of Multienzyme Complexes and Their Use in Cold Hydrolysis of Raw Starch

Valorization of Residual Agroindustrial Cakes by Fungal Production of Multienzyme Complexes and Their Use in Cold Hydrolysis of Raw Starch

Problems with detection of proteolytic microorganisms and their undesirable activities in milk

Activity modulation of microbial enzymes by llama (Lama glama) heavy-chain polyclonal antibodies during in vivo immune responses

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