Kinetic characterization of glucose aerodehydrogenase from Aspergillus niger EMS-150-F after optimizing the dose of mutagen for enhanced production of enzyme
Braz. j. microbiol
; 44(4): 1105-1112, Oct.-Dec. 2013. ilus, graf, tab
Article
in En
| LILACS
| ID: lil-705255
Responsible library:
BR1.1
ABSTRACT
In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30°C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30°C. The Michaelis-Menton constants (Km, Vmax, Kcat and Kcat/Km) were 20 mM, 45.87 U mL-1, 1118.81 s-1 and 55.94 s-1 mM-1, respectively. The enzyme was found to be thermaly stable and the enthalpy and free energy showed an increase with increase in temperature and ΔS* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness.
Key words
Full text:
1
Index:
LILACS
Main subject:
Oxidoreductases
/
Aspergillus niger
/
Mutagenesis
/
Metabolic Engineering
/
Glucose
/
Mutagens
Language:
En
Journal:
Braz. j. microbiol
Journal subject:
MICROBIOLOGIA
Year:
2013
Type:
Article
/
Project document