The Experts below are selected from a list of 189 Experts worldwide ranked by ideXlab platform
Jean-luc Gaillard - One of the best experts on this subject based on the ideXlab platform.
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the primary structure of a low mr multiphosphorylated variant of β casein in Equine Milk
Proteomics, 2007Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Laurent Miclo, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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The primary structure of a low‐Mr multiphosphorylated variant of β‐casein in Equine Milk
Proteomics, 2007Co-Authors: Laurent Miclo, Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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Multiple forms of Equine α-lactalbumin: evidence for N-glycosylated and deamidated forms
International Dairy Journal, 2004Co-Authors: Jean-michel Girardet, Antonio Silvio Do Egito, M.-a. N’negue, Sylvie Campagna, A. Lagrange, Jean-luc GaillardAbstract:Equine raw Milk whey proteins were separated by anion-exchange fast protein liquid chromatography and characterised by alkaline polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulphate PAGE, and bi-dimensional PAGE. Approximately, 1% of α-lactalbumin (α-LA) was N-glycosylated. A minor N-glycosylated form of lysozyme was also found in Equine Milk. On the other hand, two non-glycosylated α-LA isoforms with similar molecular masses (14,215±4 Da) were shown to be present. Their respective apparent isoelectric points were 5.25 and 4.94. These isoforms did not correspond to different genetic variants and were not the result of α-LA modulation by calcium ions. They corresponded rather to a non-enzymatic deamidation process of a single asparagine side-chain, the most acidic isoform being spontaneously generated from the less acidic isoform by simple incubation of α-LA at 37°C. The initial rate of this chemical degradation was 4.5 μm ammonia liberated per hour, in 150 mm sodium phosphate buffer pH 7.4 at 37°C. Deamidation induced a slight variation in secondary structure content, but no significant change in the tertiary structure of the Equine α-LA was studied by circular dichroism in the near- and far-UV regions.
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Action of plasmin on Equine β-casein
International Dairy Journal, 2003Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Chantal Poirson, Daniel Mollé, Gérard Humbert, Laurent Miclo, Jean-luc GaillardAbstract:Abstract Equine β-casein and, in a lesser extent, αs1- and κ-caseins were good substrates for plasmin in solution. The Lys47–Ile48 bond of β-casein was readily hydrolysed leading to the production of γ-like caseins of 23 kDa. These γ-like caseins were degraded by plasmin in solution subsequently to their formation. The presence of PP5-like peptides of 20 kDa associated to caseins seemed to arise from endogenous plasmin activity in Equine Milk. Their formation was, however, not obvious in solution, as the amino-terminal region of β-casein seemed to be readily hydrolysed by plasmin. γ-caseins were generated when a sodium caseinate solution was incubated at 37°C and pH 8.0 for 24 h or when caseinate was stored in a freeze-dried form at 7°C for 2 years. This might be due to the presence of low amount of endogenous plasmin associated to Equine caseinate as evidenced by assays with a specific synthetic substrate.
Laurent Miclo - One of the best experts on this subject based on the ideXlab platform.
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the primary structure of a low mr multiphosphorylated variant of β casein in Equine Milk
Proteomics, 2007Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Laurent Miclo, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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The primary structure of a low‐Mr multiphosphorylated variant of β‐casein in Equine Milk
Proteomics, 2007Co-Authors: Laurent Miclo, Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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Action of plasmin on Equine β-casein
International Dairy Journal, 2003Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Chantal Poirson, Daniel Mollé, Gérard Humbert, Laurent Miclo, Jean-luc GaillardAbstract:Abstract Equine β-casein and, in a lesser extent, αs1- and κ-caseins were good substrates for plasmin in solution. The Lys47–Ile48 bond of β-casein was readily hydrolysed leading to the production of γ-like caseins of 23 kDa. These γ-like caseins were degraded by plasmin in solution subsequently to their formation. The presence of PP5-like peptides of 20 kDa associated to caseins seemed to arise from endogenous plasmin activity in Equine Milk. Their formation was, however, not obvious in solution, as the amino-terminal region of β-casein seemed to be readily hydrolysed by plasmin. γ-caseins were generated when a sodium caseinate solution was incubated at 37°C and pH 8.0 for 24 h or when caseinate was stored in a freeze-dried form at 7°C for 2 years. This might be due to the presence of low amount of endogenous plasmin associated to Equine caseinate as evidenced by assays with a specific synthetic substrate.
Patrick F. Fox - One of the best experts on this subject based on the ideXlab platform.
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Proteomic Comparison of Equine and Bovine Milks on Renneting
Journal of Agricultural and Food Chemistry, 2013Co-Authors: Therese Uniacke-lowe, François Chevalier, Patrick F. Fox, Sonia Hem, Daniel M. MulvihillAbstract:Rennet-induced coagulation of bovine Milk is a complex mechanism in which chymosin specifically hydrolyzes kappa-casein, the protein responsible for the stability of the casein micelle. In Equine Milk, this mechanism is still unclear, and the protein targets of chymosin are unknown. To reveal the proteins involved, the rennetability of Equine Milk by calf chymosin was examined using gel free and gel based proteomic analysis and compared to bovine Milk RP-HPLC analysis of bovine and Equine Milks showed the release of several peptides following chymosin incubation. The hydrolyses of Equine and bovine casein by chymosin were different, and the major peptides produced from Equine Milk were identified by mass spectrometry as fragments of beta-casein. Using two-dimensional electrophoresis, Equine beta-casein was confirmed as the main target of calf chymosin over 24 h at 30 degrees C and pH 6.5. The gel based analysis of Equine Milk discriminated between the different individual proteins and provided information on the range of isoforms of each protein as a result of post translational modifications, as well as positively identified for the first time several isoforms of kappa-casein. In comparison to bovine Milk, kappa-casein isoforms in Equine Milk were not involved in chymosin-induced coagulation. The intensity of Equine beta-casein spots decreased following chymosin addition, but at a slower rate than bovine kappa-casein.
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Milk | Equid Milk
Encyclopedia of Dairy Sciences, 2011Co-Authors: T. Uniacke-lowe, Patrick F. FoxAbstract:This article highlights the large interspecies differences in the composition of Milk and demonstrates the similarity between the Milks of the equids. The gross composition, constituents (proteins, lipids, carbohydrates, vitamins, and minerals), and physical properties of equid Milk, particularly Equine Milk, are summarized, and many of the chemical and physicochemical properties of Equine and asinine Milk are reported.
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Milk equid Milk
Reference Module in Food Science#R##N#Encyclopedia of Dairy Sciences (Second Edition), 2011Co-Authors: T Uniackelowe, Patrick F. FoxAbstract:This article highlights the large interspecies differences in the composition of Milk and demonstrates the similarity between the Milks of the equids. The gross composition, constituents (proteins, lipids, carbohydrates, vitamins, and minerals), and physical properties of equid Milk, particularly Equine Milk, are summarized, and many of the chemical and physicochemical properties of Equine and asinine Milk are reported.
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Equine Milk proteins: Chemistry, structure and nutritional significance
International Dairy Journal, 2010Co-Authors: T. Uniacke-lowe, Thom Huppertz, Patrick F. FoxAbstract:Equine Milk has important nutritional and therapeutic properties that can benefit the diet of the elderly, convalescent or newborn. The protein content of Equine Milk is lower than that of bovine Milk but similar to that of human Milk. In this review qualitative and quantitative differences between the caseins and whey proteins of Equine, bovine and human Milk are discussed. Important biological and functional properties of specific proteins are reviewed and their significance in human nutrition considered. As well as characterizing Equine Milk proteins in the context of human nutrition and allergology, the potential industrial exploitation of Equine Milk is explored. Cross-reactivity of proteins from different species is discussed in relation to the treatment of cows’ Milk protein allergy. While there is some scientific basis for the special nutritional and health-beneficial properties of Equine Milk based on its protein composition and similarity to human Milk, further research is required to fully exploit its potential in human nutrition.
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Indigenous enzymes in Milk: Overview and historical aspects—Part 1
International Dairy Journal, 2006Co-Authors: Patrick F. Fox, Alan L. KellyAbstract:Between 1924 and about 1970, many indigenous enzymes were identified in Milk. These were important as indicators of the adequate pasteurisation of Milk (alkaline phosphatase, γ-glutamyl transferase) or of mastitis (N-acetylglucosaminidase, acid phosphatase) and some were considered to be important for the stability of Milk (superoxidase dismutase, sulphydryl oxidase). Human and Equine Milk both contain a very high level of lysozyme, which is considered to have a significant protective effect on the neonate. Progress on the isolation and characterisation of these seven enzymes first isolated in the period 1925-1970, as well as ribonuclease, aldolase and glutathione peroxidase, from the Milk of the cow and other species and their significance in Milk and dairy products is reviewed in this article.
Antonio Silvio Do Egito - One of the best experts on this subject based on the ideXlab platform.
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the primary structure of a low mr multiphosphorylated variant of β casein in Equine Milk
Proteomics, 2007Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Laurent Miclo, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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The primary structure of a low‐Mr multiphosphorylated variant of β‐casein in Equine Milk
Proteomics, 2007Co-Authors: Laurent Miclo, Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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Multiple forms of Equine α-lactalbumin: evidence for N-glycosylated and deamidated forms
International Dairy Journal, 2004Co-Authors: Jean-michel Girardet, Antonio Silvio Do Egito, M.-a. N’negue, Sylvie Campagna, A. Lagrange, Jean-luc GaillardAbstract:Equine raw Milk whey proteins were separated by anion-exchange fast protein liquid chromatography and characterised by alkaline polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulphate PAGE, and bi-dimensional PAGE. Approximately, 1% of α-lactalbumin (α-LA) was N-glycosylated. A minor N-glycosylated form of lysozyme was also found in Equine Milk. On the other hand, two non-glycosylated α-LA isoforms with similar molecular masses (14,215±4 Da) were shown to be present. Their respective apparent isoelectric points were 5.25 and 4.94. These isoforms did not correspond to different genetic variants and were not the result of α-LA modulation by calcium ions. They corresponded rather to a non-enzymatic deamidation process of a single asparagine side-chain, the most acidic isoform being spontaneously generated from the less acidic isoform by simple incubation of α-LA at 37°C. The initial rate of this chemical degradation was 4.5 μm ammonia liberated per hour, in 150 mm sodium phosphate buffer pH 7.4 at 37°C. Deamidation induced a slight variation in secondary structure content, but no significant change in the tertiary structure of the Equine α-LA was studied by circular dichroism in the near- and far-UV regions.
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Action of plasmin on Equine β-casein
International Dairy Journal, 2003Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Chantal Poirson, Daniel Mollé, Gérard Humbert, Laurent Miclo, Jean-luc GaillardAbstract:Abstract Equine β-casein and, in a lesser extent, αs1- and κ-caseins were good substrates for plasmin in solution. The Lys47–Ile48 bond of β-casein was readily hydrolysed leading to the production of γ-like caseins of 23 kDa. These γ-like caseins were degraded by plasmin in solution subsequently to their formation. The presence of PP5-like peptides of 20 kDa associated to caseins seemed to arise from endogenous plasmin activity in Equine Milk. Their formation was, however, not obvious in solution, as the amino-terminal region of β-casein seemed to be readily hydrolysed by plasmin. γ-caseins were generated when a sodium caseinate solution was incubated at 37°C and pH 8.0 for 24 h or when caseinate was stored in a freeze-dried form at 7°C for 2 years. This might be due to the presence of low amount of endogenous plasmin associated to Equine caseinate as evidenced by assays with a specific synthetic substrate.
Jean-michel Girardet - One of the best experts on this subject based on the ideXlab platform.
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the primary structure of a low mr multiphosphorylated variant of β casein in Equine Milk
Proteomics, 2007Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Laurent Miclo, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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The primary structure of a low‐Mr multiphosphorylated variant of β‐casein in Equine Milk
Proteomics, 2007Co-Authors: Laurent Miclo, Antonio Silvio Do Egito, Jean-michel Girardet, Daniel Mollé, Patrice Martin, Jean-luc GaillardAbstract:Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's Milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length Equine beta-casein (226 residues). This low-Mr variant of Equine beta-casein displays a large deletion (residues 50-181), due to a cryptic splice site usage occurring within exon 7 during the course of primary transcripts processing. The phosphorylation pattern of this Equine beta-casein variant was investigated by LC-ESI-MS and 2-DE. Seven phosphorylation forms were identified with one to seven phosphate groups with pIs ranging between 4.67 and 4.01. The major isoforms carry five and six phosphate groups.
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Multiple forms of Equine α-lactalbumin: evidence for N-glycosylated and deamidated forms
International Dairy Journal, 2004Co-Authors: Jean-michel Girardet, Antonio Silvio Do Egito, M.-a. N’negue, Sylvie Campagna, A. Lagrange, Jean-luc GaillardAbstract:Equine raw Milk whey proteins were separated by anion-exchange fast protein liquid chromatography and characterised by alkaline polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulphate PAGE, and bi-dimensional PAGE. Approximately, 1% of α-lactalbumin (α-LA) was N-glycosylated. A minor N-glycosylated form of lysozyme was also found in Equine Milk. On the other hand, two non-glycosylated α-LA isoforms with similar molecular masses (14,215±4 Da) were shown to be present. Their respective apparent isoelectric points were 5.25 and 4.94. These isoforms did not correspond to different genetic variants and were not the result of α-LA modulation by calcium ions. They corresponded rather to a non-enzymatic deamidation process of a single asparagine side-chain, the most acidic isoform being spontaneously generated from the less acidic isoform by simple incubation of α-LA at 37°C. The initial rate of this chemical degradation was 4.5 μm ammonia liberated per hour, in 150 mm sodium phosphate buffer pH 7.4 at 37°C. Deamidation induced a slight variation in secondary structure content, but no significant change in the tertiary structure of the Equine α-LA was studied by circular dichroism in the near- and far-UV regions.
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Action of plasmin on Equine β-casein
International Dairy Journal, 2003Co-Authors: Antonio Silvio Do Egito, Jean-michel Girardet, Chantal Poirson, Daniel Mollé, Gérard Humbert, Laurent Miclo, Jean-luc GaillardAbstract:Abstract Equine β-casein and, in a lesser extent, αs1- and κ-caseins were good substrates for plasmin in solution. The Lys47–Ile48 bond of β-casein was readily hydrolysed leading to the production of γ-like caseins of 23 kDa. These γ-like caseins were degraded by plasmin in solution subsequently to their formation. The presence of PP5-like peptides of 20 kDa associated to caseins seemed to arise from endogenous plasmin activity in Equine Milk. Their formation was, however, not obvious in solution, as the amino-terminal region of β-casein seemed to be readily hydrolysed by plasmin. γ-caseins were generated when a sodium caseinate solution was incubated at 37°C and pH 8.0 for 24 h or when caseinate was stored in a freeze-dried form at 7°C for 2 years. This might be due to the presence of low amount of endogenous plasmin associated to Equine caseinate as evidenced by assays with a specific synthetic substrate.