We isolated thirty thermophilic fungi with ability to grow at 45 °C on medium containing wheat bran and casein as carbon sources and produced proteases in both SSF and SmF (data not shown). The fungus strain F.2.1.4 presented the highest protease activity and was sub-jected to taxonomic characterization.
Analyses of macro and micro-morphological characteristics accommodated the isolate F.2.1.4 in the genus Myceliophthora due to the presence of spreading colonies with dense aerial mycelium and blastic conidia often borne in ampulliform swellings (Van Oorschot, 1980). Particularly, this isolate produced pulverulent pale mycelium, obovoid conida measuring 5.65-7.91 x 3.39-4.52 μm as well as conidia walls finely ornamented. Based on such
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Alkaline proteases have many applications in the industry such as laundry detergents, leather processing, brewing, food and pharmaceutical industries therefore the alkalophilic properties exhibited by proteases from Myceliophthora sp. in SSF are an important aspect to be explored.
The maximum activity for both crude enzymes was 50ºC but the crude enzyme from SSF maintained 95% of the maximum activity at 60ºC while for the protease from SmF the activity was 79% (Figure 3b). This property is consistent with the dates reported for extracellular enzymes from thermophiilic fungi. Similar results were reported for proteases from Thermoascus aurantiacus var levisporus (55oC) (Marcy et al., 1984). Proteases from thermophilic Scytalidium (Hasbay and Ögel, 2002) and from thermotolerant Aspergillus fumigatus (Santos et al., 1996) showed optima activities at 45 oC.
Since the protease activity was higher in crude enzyme solution from SSF and addi-tionally, it showed alkaline optimum pH and activity at higher temperature range (50-65 ºC), it was used for the immobilization procedure.
Entrapment of the protease in calcium alginate beads
Among the quantities of sodium alginate (2, 2.5, 3, 3.5 % (w/v)) and CaCl2 concentrations (0.04, 0.06, 0.08, 0.1 M), the best condition for immobilization of the protease was 2.5 % alginate and 0.08 mM CaCl2 (data not shown), obtaining 2 mm (average diameter) beads.
The immobilized enzyme was reevaluated
In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or
Of the thousands of enzymes known, there is a family of enzymes called proteases that catalyze a reaction of breaking down proteins. What do you think would happen if you added a protease to your sample of catalase before proceeding with your experiment?
The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the
Enzymes are types of proteins that work as a substance to help speed up a chemical reaction (Madar & Windelspecht, 104). There are three factors that help enzyme activity increase in speed. The three factors that speed up the activity of enzymes are concentration, an increase in temperature, and a preferred pH environment. Whether or not the reaction continues to move forward is not up to the enzyme, instead the reaction is dependent on a reaction’s free energy. These enzymatic reactions have reactants referred to as substrates. Enzymes do much more than create substrates; enzymes actually work with the substrate in a reaction (Madar &Windelspecht, 106). For reactions in a cell it is
The colonies were smooth, translucent, and had a white brownish color. The Gram stain resulted in Gram positive cocci. After the Gram stain was completed, the bacteria were streaked on a Mannitol-Salt Agar plate and a Catalase test was performed. After these test were completed a Phenol Red Dextrose Fermentation tube was inoculated, and a SIM Tube inoculated.
These results show how temperature of extreme high, or low affects enzyme activity. The highest rate of enzyme activity occurred at 37 Cº. Anything that was hotter or cold than 37 Cº slowed the reaction rate. As I thought, 100 degrees would denature the enzyme, and that was the case. The data provided shows exactly what temperatures enzymes work best, and worst. The objective was achieved as we discovered the different reaction rates under different temperatures. The results are reliable, as we know enzymes do not work well when under extreme heat or denaturation occurs. What I learned in this experiment was that enzymes don’t work well under cold temperatures because they tend to move slower. My hypothesis did not quite match, because I thought they work best at lower temperatures.
Most enzymes work best at body temperature, higher temps will cause the enzyme to no longer work properly
If temperature of the water(enzyme environment) is increased to 35°C, then the enzyme activity will
5. Heat treatment was used to denature the sucrase that was added to the control test tube. In the experimental test tube, alkaline
according to the temperature but it will be faster in bacterial than in fungal, 100% hydrolysis
The purpose of this study is to identify four unknown organisms. The unknown organisms have been assigned randomly to six-research groups by Professor Hoffman. Each research group was provided two eukaryotes and two prokaryotes. The unknown organisms will fall into the following classifications: bacteria, algae, fungi, or protozoans. All living organisms are organized into one of three domains of life, Bacteria, Archaea, and Eukarya.
In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the
During these experimental procedures, the implication of multiple different temperatures on fungal and bacterial amylase was studied. In order to conduct this experiment, there were four different temperatures used. The four temperatures used were the following: 0 degrees Celsius, 25 degrees Celsius, 55 degrees Celsius, and 80 degrees Celsius - Each temperature for one fungal and one bacterial amylase. Drops of iodine were then placed in order to measure the effectiveness of the enzyme. This method is produced as the starch test. The enzyme was tested over the course of ten minutes to determine if starch hydrolysis stemmed. An effective enzyme would indicate a color variation between blue/black to a more yellowish color towards the end of the time intervals, whereas a not so effective enzyme would produce little to no change in color variation. According to the experiment, both the fungal amylase and bacterial amylase exhibited a optimal temperature. This was discovered by observing during which temperature and time period produced a yellow-like color the quickest. Amylase shared a similar optimal temperature of 55 degrees Celsius. Most of the amylases underwent changes at different points, but some enzymes displayed no effectiveness at all. Both amylases displayed this inactivity at 0 degrees Celsius. At 80 Celsius both the enzymes became denatured due to the high temperatures. In culmination, both fungal and bacterial amylase presented a array of change during it’s
This experiment was conducted to test the properties of enzymes. Enzymes significantly reduce the activation energy reactions by binding to substrate molecules. Enzymes function optimally due to the temperature and pH. At certain temperatures and pH, bonds such as hydrogen and ionic are broken and this allows an enzyme to bind to its substrate. The proteolytic enzyme trypsin was used for this experiment. More about function of this enzyme is included in the lab manual.
In triplicates of Pleurotus pulmonarius, white mycelium colonies were formed in both MEA and PDA media, but white and orange colony were observed in GYMP medium. The MEA medium causes Pleurotus pulmonarius to form dense mycelia, but less dense in media PDA and GYMP. Totally, none of the media contributed to sporulation from Pleurotus pulmonarius and moreover, in MEA medium, it took 6 days for it to grow to the maximum length provided while the GYMP medium, it took 7 days. As for the PDA medium, even after 7 days, it did not reach the maximum length of the petri dish.