This study focused on the determination of non-specific proteolytic activity of edible spruce Morchella esculenta in water extract, phosphate-buffered saline (PBS) solution (pH = 7.5) extract and a suspension prepared from 200 mg DW (dry weight) of edible spruce in PBS solution (pH 7.5). A clear casein solution was used as a substrate. The absorbances were measured in quartz cuvettes at the wavelength of 280 nm against a blank with zero concentration of trypsin. Non-specific proteolytic activity was expressed as trypsin equivalents per kilogram of mushroom dry weight (mg.kg−1 DW). All of the extracts demonstrated non-specific enzymatic activity. The highest activity was observed in the PBS suspension and the lowest enzymatic activity was measured in the water extract of the Morchella esculenta fungi. The non-specific proteolytic activity decreased in the following order: PBS suspension extract (pH 7.5; 22.9 mg.kg−1 DW), followed by PBS extract (pH 7.5; 13.6 mg.kg−1 DW) and finally the water extract (10.94 mg.kg−1 DW).
, J.M. (2005): Cloning and characterisation of an aspartyl protease inhibitor (API-1) from Ancylostoma hookworms. Int. J. Parasitol., 35: 303 - 313. DOI: 10.1016/j.ijpa- ra.2004.11 014 DUBOVSKAYA, A.YA. (1973): A study of proteolyticactivity in some cestode species. Parazitologiya, 7:154 -159 (In Russian) DZIK, J.M. (2006): Molecules released by helminth parasites involved in host colonization. Ada Biochim. Pol., 53(1): 33 - 64 HALTON, D.W. (1997): Nutritional adaptations to parasitism within the Platyhelminthes. Int. J. Parasitol.,27:693-704. DOI: 10.1016/ S0020
The leaves of Zantedeschia and Hosta are used as florist greens in different floral arrangements. The most efficient postharvest treatment for cut foliage is the use of growth regulators, which prolong their vase life by delaying degradative changes occurring in leaves, especially proteolysis. Cycloheximide (CHI) is one of the protein synthesis inhibitors, blocking the enzymes responsible for decreasing membrane integrity, a phenomenon hastening senescence. The aim of this experiment was to evaluate the effects of CHI and benzyladenine (BA) or gibberellic acid (GA3) on the longevity of cut foliage in hosta (Hosta sp.) cultivars and Ethiopian calla (Zantedeschia aethiopica) and to follow the changes in certain proteolytic processes occurring during senescence. Generally, 24 h conditioning with cycloheximide shortened the longevity of cut calla leaves while having no effect on hosta vase life. In ageing leaves of ‘Minima Glauca’ hosta and calla, the total proteolytic activity increased, including that of cysteine protease. Due to the application of BA or GA3 in hosta and calla, respectively, this activity was limited. On the contrary, the use of CHI either did not affect the activity of cysteine protease or increased it several-fold relative to the control, in hosta and calla, respectively. Leaves treated with growth regulators had many more soluble proteins and fewer free amino acids, including free proline, than leaves from other treatments. The highest free proline level was determined in calla leaves conditioned with CHI, where it increased 18-fold relative to the initial level.
., Podestà A. (2004) Expression profile of water-soluble proteinases during ontogenesis of Megachile rotundata: an electrophoretic investigation. Apidologie 35(6): 595-604. Frączek R. J., Żółtowska K., Lipiński Z., Dmitryjuk M. (2013) The mutual influence of proteins from Varroa destructor extracts and from honeybee haemolymph on their proteolyticactivity - in vitro study. Acta Parasitologica 58(3): 317-323. Giejdasz K., Wilkaniec Z. (2002) Individual development of the red mason bee (Osmia rufa L., Megachilidae) under natural and laboratory conditions. Journal of
enzymatic in vitro assay with the caspase 3-specifc substrate Ac-DEVD-pNA. The results showed very low activity (0.18 ± 0.06 μmol pNA min -1 mg -1 ) at initial stage, in microspores before the inductor stress, whereas it greatly increased (1.28 ± 0.13 μmol pNA min -1 mg -1 ) in stress-treated microspores ( Fig. 4 ). Controls with the caspase-specific inhibitor Ac-DEVD-CHO readily inhibited proteolyticactivity of caspase 3 in both stages analysed of B. napus microspore embryogenesis, supporting the specificity of the enzymatic assay. Figure 4 Caspase 3-like activity
References ADAV, S.S., LEE, D.J., LAI, J.Y.: Proteolyticactivity in stored aerobic granular sludge and structural integrity. Bioresour. Technol., 100, 2009, 100, 68-73. BARUZZI, F., LAGONIGRO, R., QUINTIERI, L., MOREA, M., CAPUTO, L.: Occurrence of non-lactic acid bacteria populations involved in protein hydrolysis of cold-stored high moisture Mozzarella cheese. Food Microbiol., 30, 2012, 37-44. BAUR, C., KREWINKEL, M., KRANZ, B., VON NEUBECK, M., WENNING, M., SCHERER, S., STOECKEL, M., HINRICHS, J., STRESSLER, T., FISCHER, L.: Quantification of the proteolytic
Evaluation of the enzymatic activity of selected bacterial strains
In these studies we attempted to evaluate the lipolytic, proteolytic and cellulolytic activity of bacterial strains isolated from water and the bottom sediments of Turawa Lake. The following bacterial genera prevailed among the isolated strains: Bacillus, Pseudomonas, Enterobacter, Cellulomonas and Cytophaga. The lipolytic activity was determined using a titrimetric method, the proteolytic activity — using a modified Anson method, and the cellulolytic activity - on the basis of mass decrement of a cellulose disk after 14 days of bacterial culture. The cultures were maintained at 28°C, pH 7.0 with the following substrates: olive oil, albumin and cellulose disk. Among the analysed microorganisms, Bacillus and Pseudomonas strains showed the highest lipolytic and proteolytic activity. In the cellulolytic assay Cytophaga bacteria showed about twofold higher activity than that of Celulomonas.
Background. Tumor tissue-associated KLKs (kallikrein-related peptidases) are clinically important biomarkers that may allow prognosis of the cancer disease and/or prediction of response/failure of cancer patients to cancerdirected drugs. Regarding the female/male reproductive tract, remarkably, all of the fifteen KLKs are expressed in the normal prostate, breast, cervix uteri, and the testis, whereas the uterus/endometrium and the ovary are expressing a limited number of KLKs only.
Conclusions. Most of the information regarding elevated expression of KLKs in tumor-affected organs is available for ovarian cancer; depicting them as valuable biomarkers in the cancerous phenotype. In contrast, for breast cancer, a series of KLKs was found to be downregulated. However, in breast cancer, KLK4 is elevated which is also true for ovarian and prostate cancer. In such cases, selective synthetic KLK inhibitors that aim at blocking the proteolytic activities of certain KLKs may serve as future candidate therapeutic drugs to interfere with tumor progression and metastasis.
Fetuin-A is a secretory liver glycoprotein with multiple physiological functions such as regulation of insulin resistance, tissue calcification, bone metabolism, cellular proteolytic activity, and self-proliferative signaling.
Fetuin-A is a unique molecule which binds to the insulin receptor, modulating its sensitivity, and transducing “the physiological conditions” (serum levels of the metabolites like glucose, free fatty acids, inflammatory signals) from outside into inside the cells. Plasma fetuin-A levels correlate with reduced glucose tolerance and insulin resistance. Impaired insulin sensitivity leads to the development of metabolic syndrome, an increased risk for type 2 diabetes (T2DM), dyslipidaemias and cardiovascular diseases (CVDs). Furthermore, fetuin-A inversely correlates with inflammatory and activation biomarkers, e.g. in patients with T2DM. Thus, circulatory fetuin-A levels may have plausible predictive importance as a biomarker of risk of diabetes and negative acute phase protein. Dysregulated, it plays a crucial role in the pathogenesis of some metabolic disorders and clinical inflammatory conditions like metabolic syndrome, T2DM, CVDs, polycystic ovary syndrome (PCOS), etc.