Paulo Roberto Martins Bonilha (2006) et. al... Cyclodextrin glycosyltransferase (EC
2.4.1.19) is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. An alkalophilic Bacillus strain, isolated from cassava peels, was identified as Bacillus licheniformis. CGTase production by this strain was better when potato starch was used as carbon source, followed by cassava starch and amylopectin. Glucose and amylose, on the other hand, acted as synthesis repressors. When the cultivation was supplemented with sodium ions and had the pH adjusted between 6.0 and 9.0, the microorganism maintained the growth and enzyme production capacity. This data is interesting because it contradicts the concept that alkalophilic microorganisms do not grow in
…show more content…
TPR71H. From this study we found that the optimized parameters for the production of
CGTase by Bacillus sp. TPR71H were Soluble Starch 3%, Yeast Extract 0.5%, K2HPO4 0.1%,
Inoculum Level 3.5%, Inoculum Age 24h, Incubation Period 36h, rpm 220, Incubation
Temperature 32°C and the pH of 7.5.
M. Manoj Saravana Guru (2012) et. al.. Cyclodextrin glucosyltransferase (CGTase) producing alkalophilic bacteria were isolated from the water samples collected from Marneri pond of Tirunelveli, Tamil Nadu by serial dilution and plating method. Totally 22 bacteria were isolated from the collected water samples and screened for CGTase activity by Horikoshis medium II agar plate method. Among 22 bacterial isolates, 15 isolates showed CGTase activity and better zone producing strain was selected for further studies. The potential strain was identified as Bacillus circulans by 16S rDNA gene sequencing experiment. The best enzyme activity was observed at pH 10.5, 32°C, supplemented with cassava as carbon source an d beef extract as nitrogen source. CGTase was purified to 20.21 fold with a yield of 55.14% recording
The tube was placed back in incubation for 96 more hours to observe any more positives. 2.10 Catalase Test A trypticase soy agar plate was used and after incubation, four drops of 3% Hydrogen Peroxide was added to the plate to flow over the bacterial growth. A presence of bubbling was observed. 2.11 Starch Hydrolysis
By completing this experiment, knowledge collected about optimal pH in enzymes will help
Sucrase activity increases with increasing sucrose concentration Materials and Methods Effect of pH on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2. Independent Variable pH 3. Controlled Variables temperature, amount of substrate (sucrose) present, sucrase + sucrose incubation time Effect of Temperature on Enzyme Activity 1.
Exercise 14: Unknown Identification Lab Report The purpose of the study was to identify the unknown bacterium using various biochemical tests in addition to using scientific methods in determining the outcome of the hypothesis. Each biochemical test will help determine the bacteria based on specific characteristics of each organism. I was giving unknown number 232. The first procedure that needed to be done after obtaining unknown bacterial mixture was to isolate the two bacteria in a pure culture using the streak plate method described in Microbiology Laboratory Manual Eight Edition. The material used was trypticase soy agar (TSA) plate, nutrient plate, starch agar, hydrogen peroxide, iodine reagent and microscope.
Materials and methods Isolation and identification of Bacillus thuringiensis Bacillus thuringiensis was isolated from raw milk (Taif, KSA) on nutrient agar at 37oC for overnight. The supernatant of the bacterial isolate was screened for synthesis of AgNPs. The bacterial isolate was morphologically and biochemically characterized according to Bergy’s Manual of Systemic Bacteriology[21]. Also, this bacterial isolate was further identified by 16S rRNA sequencing.
Title: How Ph Levels Affected the Fermentation of Beer Hypothesis: The beer will be left with more sugar deposit as the Ph levels increase because alpha/beta -amylase will no longer function. Predictions: Alcohol Percentage Analysis for the Control and the Experimental During this experiment, the pH level was increased, therefore Alpha-Amylase was favored. Due to the nature of Alpha-Amylase cutting randomly through a large carbohydrate molecule, it leaves bigger sugars in the flask, which cannot be digested by yeast. Due to this, less reactions should occur in the experimental, therefore leading to a lower percentage of alcohol production, compared to the control.
1.1 Abstract The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample. The purpose of stopped enzyme assay to study B-galactosidase is to determine the effect of temperature and concentrations of substrate on enzyme activity.
The B. Vulgaris samples were approximately 1cm3. They were kept the same size to ensure accurate results. A control test was conducted in distilled water to obtain a result to compare. The ethanol treatments were 40% and 70%. To prepare the solutions a 70% ethanol solution was used to make 40%.
In this experiment , we can prove that the temperature, pH and salt are the factors that will affect the structure and function of the enzyme as it is a kind of protein . Therefore, there may be an influence on the activity of enzyme which substrates cannot be binded on the active site if the amylase in too high or low ph and temperature and excess salt environment . On the other hand optimum ph and temperature and suitable salt concentration may favour the amylase activity . Reference : 1.2016, May 08). Effects of pH on Amylase Activity.
Introduction 1.1 Aim: To determine the kinetic parameters, Vmax and Km, of the alkaline phosphatase enzyme through the determination of the optimum pH and temperature. 1.2 Theory and Principles (General Background): Enzymes are highly specific protein catalysts that are utilised in chemical reactions in biological systems.1 Enzymes, being catalysts, decrease the activation energy required to convert substrates to products. They do this by attaching to the substrate to form an intermediate; the substrate binds to the active site of the enzyme. Then, another or the same enzyme reacts with the intermediate to form the final product.2 The rate of enzyme-catalysed reactions is influenced by different environmental conditions, such as: concentration
Introduction: In this task I will be researching the effect that acid rain has on the rate of plant growth. Acid rain is any type of precipitation with a high pH, with high levels of nitric acids. The reason why I had chosen this topic was because acid rain seems to have a great effect on the effect of plant growth, and plants play a very important role in our ecosystem. Acid rain is a major problem in our environment when we are not able to neutralize the acidity.
Along with being found in plants, they are also present in liver cells, kidney cells, leukocytes and erythrocytes. For the concentration of enzyme experiment, the hypothesis was if the concentration of an enzyme increases, then the enzyme activity will increase as well. The hypothesis was proven to be true, because there are more enzymes to react with substrates. For the enzyme—factors affecting, the hypothesis concluded was if the temperature increases, than the enzyme activity will increase. This however was proven wrong, because enzymes become unstable at higher temperatures.
Usually, the microbial enzymes have various potential uses in industries and medicine. The microbial enzymes are also more reliable than plant and animal enzymes as they are more stable and active. Also the microorganisms demonstrate an alternative source of enzymes because they can be cultured in large quantities in a short time by fermentation and owing to their biochemical diversity and susceptibility to gene manipulation. Industries are looking for new microbial strains in order to produce different enzymes to fulfil the current enzyme
In order to utilize casein, bacteria cells secrete proteolytic exoenzymes (amylases, proteases, pectinases, lipases, xylanases and cellulases) outside of the cell that hydrolyze the protein to amino acids. The amino acids can then be used by cells after crossing the cell membrane via transport proteins [169]. Starch hydrolysis test is used to differentiate bacteria based on their ability to hydrolyze starch with the enzyme α-amylase or oligo-l, 6-glucosidase. These enzymes hydrolyze starch by breaking the glycosidic linkages between the sugar subunits. It aids in the differentiation of species from the genera Corynebacterium, Clostridium, Bacillus, Bacteroides, Fusobacterium and members of Enterococcus [170].
The media used in this experiment was Trypticase nitrate broth. The reagents used (A and B) were sulfanilic acid and alpha-naphthylamine (respectively). Using aseptic technique, the bacterium (16A and 16B) were inoculated into labeled broth test tubes. The tubes were incubated for 48 hours at 37 degrees Celsius. When the incubation was complete 5 drops of reagent A and 5 drops of reagent B were added to each of the broths.