A simple and rapid assay of collagen-like polymer in crude lysate from Escherichia coli

Yin, Jia; Tomycz, Luke; Bonner, Gary; Wang, Daniel I. C.

doi:10.1016/s0167-7012(01)00372-4

Cited by 10 publications

(7 citation statements)

References 8 publications

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“…Thus, particular care should be taken when using this method on semipurified extracts, with particular emphasis placed on blank measurements (Lim et al, 1993). Despite this limitation, this assay remains a relatively simple method to perform and a number of studies have successfully used it with minor modifications of the incubation time and/or the amount of the substrate to determine collagenases isolated from a variety of sources (Rosen, 1957;Yoshida and Noda, 1965;Endo et al, 1987;Sivakumar et al, 1999;Yin et al, 2002;Park et al, 2002;Wu et al, 2010). …”

Section: Colorimetric Assaysmentioning

confidence: 99%

Extraction and Purification of Collagenase Enzymes: A Critical Review

Daboor¹,

Budge²,

Ghaly

et al. 2010

American Journal of Biochemistry and Biotechnology

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Problem statement: Enzymes have vital roles in several industrial processes (foods, cosmetics, nutraceuticals and pharmaceuticals) due to their highly selective nature and high activity at very low concentrations. Recent efforts to identify new sources of useful enzymes have been concentrated on the marine environment because of the potential to make use of processing wastes. About 35-50% of the mass of the fish caught is a waste that is disposed off at sea or in landfills. The extraction of enzymes from fish processing waste can reduce environment problems and improve the economics of the fish industry. Collagenases are a group of enzymes that can be extracted from fish waste. Approach: Comprehensive reviews of the literature on the extraction, purification, characterization and use of collagenases was carried out. Results: Collagenases have different molecular weights based on their types and sources. They have the ability to break down the peptide bonds in collagen at physiological pH. They are classified into two types: serine and metallocollagenase. Collagenolytic activities have been shown at a wide range of temperatures (20-40°C) and pH (6-8). Many activators can be used to achive collagenase activity including 4-Aminophenylmercuric Acetate (APMA), trypsin, potassium or sodium thiocyanate, iodoacetamide and potassium iodide. Dithiothreitol (DTT), mercaptoethanol, ethylendiaminetetracetic acid, ophenanthroline and cysteine inactivate the enzyme. Collagenases enzymes can be extracted with a variety of techniques using different buffering systems (tris-HCl, sodium bicarbonate, calcium chloride and cacodylate). All techniques involve the use of ammonium sulphate fractionation and centrifugation to precipitate the enzyme. Collagenases are normally purified using chromatographic techniques such as gel-filtration, ion-exchange and affinity column chromatography. Collagenase can be assayed with a number of methods, including: colorimetric absorbance, viscometry, radioactivity and fluorescence spectroscopy. Collagenases are partly responsible for toughness in red meats and are used as tenderizers in food industry, have application in the fur and hide tanning to ensure uniform dying of leather, used in medicine to treat burns and ulcers, eliminate scar tissues, transplantation of organs. Conclusion: Understanding of the nature of the enzymes and identifying the most suitable resources and the methods for their extraction and purification will have significant impact on the fish processing, food and medical industries.

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Section: Colorimetric Assaysmentioning

confidence: 99%

Extraction and Purification of Collagenase Enzymes: A Critical Review

Daboor¹,

Budge²,

Ghaly

et al. 2010

American Journal of Biochemistry and Biotechnology

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“…Collagen-like polymer (CLP) was measured using the method of Yin et al (2002). Tissues were homogenized in buffer (5 mM CaCl 2 , 20 mM Tris-HCl, pH 7.5) and tissue debris was then removed by centrifugation at 12,000 rpm for 20 min.…”

Section: Collagen-like Polymer Assaymentioning

confidence: 99%

“…In contrast to natural collagen with forms of three lefthanded helical polypeptide chains, CLP is a denatured product and only has one polypeptide chain (Yin et al, 2002). To detect increased collagen, we measured the CLP levels in injured tissues.…”

Section: Effect Of Coq10 On Collagen-like Polymer (Clp) Level In Skinmentioning

confidence: 99%

Effect of coenzyme Q10 on cutaneous healing in skin-incised mice

et al. 2009

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Coenzyme Q10 (CoQ10) is a biosynthesized quinone with 10 isoprene side chains in humans. To investigate the anti-inflammatory and wound healing effect of CoQ10, we performed in vivo and in vitro experiments. In vivo studies, there were 3 groups; Naive (without skin incision), Control (with skin incision) and CoQ10 (100 mg/kg treatment with skin incision). Collagen-like polymer (CLP) level of CoQ10 group was increased significantly compared to the control group (p<0.05). Also, CoQ10 group showed significant inhibition on myeloperoxidase (MPO) and PLA(2) level compared to the control group (p<0.05). These data show that CoQ10 may have an anti-inflammatory and a wound healing effect. CoQ10 showed significant antioxidant activity in vivo on malondialdehyde (MDA) and superoxide dismutase (SOD) levels compared to the control group (p<0.05). Although CoQ10 did not show antioxidant activity in cell free system of DPPH radical scavenge, it had a potent antioxidant activity in cell culture system of both silica- and zymosan-induced reactive oxygen species generation using Raw 264.7 cells. This result may be associated with the conversion of CoQ10 to the reduced form (CoQ10H(2)) in the presence of some kinds of intracellular reducing agents. In conclusion, it is considered that CoQ10 appears to have a cutaneous healing effect in vivo, which may be related to the secondary action of CoQ10.

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“…Collagen-like polymer (CLP) was assessed using the method of Yin et al (2002). Tissues or NIH 3T3 cells were homogenized in buffer (5 mM CaCl2, 20 mM Tris-HCl, pH 7.5) and tissue debris was then removed by centrifugation at 12,000 rpm for 20 min.…”

Section: Collagen-like Polymer Assaymentioning

confidence: 99%