Aspergillus can be used to produce α-amylase (α-AM), which is an enzyme that hydrolyzes the glycosidic bonds in starch molecules into maltose (disaccharide). Pure α-AM has a specific activity of 56.00 U/mg. The α-AM is secreted into the culture media during production (after 5 days, production is complete). Following, the Aspergilluscells are separated by centrifugation and discarded. The culture supernatant (with α-AM) is filtered giving a clarified supernatant with a volume of 100.0 L, activity of 3.00 U/mL, and total protein concentration of 0.650 mg/mL. After purification, the product stream (7.50 L) contains α-AM (25.00 U/mL) and a total of 6.788 x 103 mg total protein. Calculate the following: Concentration of total protein (g/L) and total units of activity in the clarified supernatant and purification product stream. Specific activities (U/g) of the clarified supernatant and purification product stream. Consider significant figures in your answers (may want to use scientific notation to assist). α-AM purities in the clarified supernatant and purified product stream. Fold-purification. Concentration factor and yield of α-AM for the purification step.
Microscopic examination
The analysis of minute organisms, cellular organization of any biological structure, and composition of body fluids with the help of a microscope is known as microscopic examination. The magnification of specimens or samples under study helps in attaining a clearer picture of it.
Gram Staining
Named after Hans Christian Gram, a Danish bacteriologist, Gram stain is one of the most powerful staining techniques within microbiology. This technique was introduced in 1882 to identify pneumonia-causing organisms. The Gram staining technique uses crystal violet or methylene blue as primary staining colors to distinguish gram-positive from gram-negative organisms. Under a microscope, the gram-positive organisms appear purple-brown, retaining the primary color. Gram-negative organisms appear pink or red as they do not acquire the color of the primary stain.
- Aspergillus can be used to produce α-amylase (α-AM), which is an enzyme that hydrolyzes the glycosidic bonds in starch molecules into maltose (disaccharide). Pure α-AM has a specific activity of 56.00 U/mg. The α-AM is secreted into the culture media during production (after 5 days, production is complete). Following, the Aspergilluscells are separated by centrifugation and discarded. The culture supernatant (with α-AM) is filtered giving a clarified supernatant with a volume of 100.0 L, activity of 3.00 U/mL, and total protein concentration of 0.650 mg/mL. After purification, the product stream (7.50 L) contains α-AM (25.00 U/mL) and a total of 6.788 x 103 mg total protein. Calculate the following:
- Concentration of total protein (g/L) and total units of activity in the clarified supernatant and purification product stream.
- Specific activities (U/g) of the clarified supernatant and purification product stream. Consider significant figures in your answers (may want to use scientific notation to assist).
- α-AM purities in the clarified supernatant and purified product stream.
- Fold-purification.
- Concentration factor and yield of α-AM for the purification step.
Alpha amylase is an enzyme that hydrolyzes the alpha-1,4 glycosidic bonds in starch molecules, breaking them down into smaller carbohydrates. This enzyme is widely used in various industries, including food, brewing, and biofuel production. It is produced by several microorganisms, including Aspergillus, and has a specific activity that can be measured in units per milligram.
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