Optimization of protease production from surface-modified coffee pulp waste and corncobs using Bacillus sp. by SSF.
The aim of the study was to identify new sources of substrate from agro-industrial waste for protease production using Bacillus sp., a local bacteria isolated from an agro-waste dumping site. The strain was identified as Bacillus sp. BT MASC 3 by 16S rRNA sequence followed by phylogenic analysis. Response surface methodology-based Box-Behnken design (BBD) was used to optimize the variables such as pH, incubation time, coffee pulp waste (CPW) and corncob (CC) substrate concentration. The BBD design showed a reasonable adjustment of the quadratic model with the experimental data. Statistics-based contour and 3-D plots were generated to evaluate the changes in the response surface and understand the relationship between the culture conditions and the enzyme yield. The maximum yield of protease production (920 U/mL) was achieved after 60 h of incubation with 3.0 g/L of CPW and 2.0 g/L of CC at pH 8 and temperature 37 °C in this study. The molecular mass of the purified enzyme was 46 kDa. The highest activity was obtained at 50 °C and pH 9 for the purified enzymes.
Centre for Biotechnology, Muthayammal College of Arts and Science, Rasipuram, Namakkal, Tamil Nadu, 637 408, India.
Selvam
S
Kandasamy
Department of Applied Sciences, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, Hokkaido, 050-8585, Japan.
Govarthanan
G
Muthusamy
Centre for Biotechnology, Muthayammal College of Arts and Science, Rasipuram, Namakkal, Tamil Nadu, 637 408, India. nbsenthilkumar@gmail.com.|Department of Medical Microbiology, College of Health and Medical Sciences, Haramaya University, P.O. Box 235, Harar, Ethiopia. nbsenthilkumar@gmail.com.
Senthilkumar
S
Balakrishnan
Department of Marine Biotechnology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620 024, India.
Senbagam
S
Duraisamy
PG and Research Department of Biotechnology, Mahendra Arts and Science College (Autonomous), Kalippatti, Namakkal, Tamil Nadu, 637501, India. t_selvankumar@yahoo.com.
Selvankumar
S
Thangasamy
Division of Biotechnology, Advanced Institute of Environment and Bioscience, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, 570752, South Korea.
Kamala-Kannan
KK
Seralathan
PG and Research Department of Biotechnology, Mahendra Arts and Science College (Autonomous), Kalippatti, Namakkal, Tamil Nadu, 637501, India.
Sudhakar
S
Chinnappan
Bacillus sp.; Coffee pulp waste; Corncobs; Protease; Response surface methodology; SSF