Kinetics of alkaline protease production by Streptomyces griseoflavus PTCC1130.
OBJECTIVE
Proteases are a group of enzymes that catalyze the degradation of proteins resulting in the production of their amino acid constituents. They are the most important group of industrial enzymes which account for about 60% of total enzymes in the market and produced mainly by microorganisms. The attempts were made to study the kinetic parameters of protease produced by Streptomyces griseoflavus PTCC1130.


METHODS
Streptomyces griseoflavus PTCC1130 was grown on casein agar. Different media such as BM1, BM2, BM3 and BM4 were prepared. Data obtained from growth and protease production were subjected to kinetics evaluation. Casein was used as substrate for protease activity and the released soluble peptide bearing aromatic amino acid were quantified by Folin Cioclateaue reagent. Protein content of the enzyme and the sugar utilized by the organism were estimated by Bradford and Miller's methods respectively.


RESULTS
Basal Medium named as BM1, BM2, BM3 and BM4(50 mL in 250 mL Erlen Meyer flasks) were screened out to evaluate protease production by Streptomyces griseoflavus PTCC1130. They were inoculated with known amount of seed culture and kept on rotary shaker. To obtain the specific growth rate, wet weight of biomass was plotted against the time. The clarified supernatant was used for the analysis of protease by measuring the soluble peptide containing aromatic amino acid residues employing Folin Cioclateaue reagent. Our results showed that maximum level of enzyme production (14035 U/L) was occurred at late exponential phase using Basal Medium supplemented with zinc sulfate (0.5g/L), casein (10g/L) at pH 6.5.


CONCLUSIONS
A kinetic study of protease production by Streptomyces griseoflavus PTCC1130 provided highly quantitative information regarding the behavior of a system, which is essential to study the fermentation process. Exploitation of such kinetics analysis would be useful in commercialization of microbial enzyme production.
Chemical Engineering Department, Islamic Azad University, Shah Rood Branch, Shah Rood, Iran.
Seyed Vesal
SV
Hosseini
Pilot Nanobiotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran.
Zahra
Z
Saffari
Pilot Nanobiotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran.
Ali
A
Farhanghi
Pilot Nanobiotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran.
Seyed Mohammad
SM
Atyabi
Pilot Nanobiotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran.
Dariush
D
Norouzian
Kinetics of fermentation; Microbial enzyme; Protease; Streptomyces griseoflavus