Evaluation of the Effects of Nutritional and Environmental Parameters on Extracellular Protease Production by Stenotrophomonas acidaminiphila Strain BPE4

An aerobic mesophilic Gram-negative rod-shaped bacterium isolated from fermenting bean processing effluent (BPE) and identified as Stenotrophomonas acidaminiphila strain BPE4, produced an extracellular protease on skimmed-milk minimal medium. Time-course of enzyme production revealed peak productivity at 12 h but enzyme concentration gradually increased till 36 h beyond which both concentration and productivity gradually decreased. Evaluation of the influences of major nutritional sources on enzyme concentration revealed significant (P < 0.05) effects of fermentation time and nutrient sources, with corn steep liquor and NH4Cl emerging as best carbon and nitrogen sources respectively. The study also selected K2HPO4/KH2PO4 (2:1) and 108 cfu/mL as most appropriate phosphate combination and inoculum size respectively for maximum release of protease within the fermentation time of 36 h. One unit of proteolytic activity was defined as the amount of crude enzyme that digested 1 mg of azocasein in one minute under the assay conditions. Assessment of optimum conditions of temperature and pH for test enzyme activity revealed optimum enzyme activity of 158.83 U/mL (2647.70 nkatal) at 40°C and pH 9.0 suggesting protease of the alkaline kind. The enzyme was specific for casein as a protein substrate. Stenotrophomonas acidaminiphila strain BPE4 could be exploited for commercial production of extracellular protease on corn steep liquor as a waste management option and industrial applications.

[1]  S. Antai,et al.  Selection of Enterobacter cloacae Strain POPE6 for Fermentative Production of Extracellular Lipase on Palm Kernel Oil Processing Effluent , 2017 .

[2]  S. Antai,et al.  Water Soluble Fraction of Crude Oil Uncouples Protease Biosynthesis and Activity in Hydrocarbonoclastic Bacteria; Implications for Natural Attenuation , 2017 .

[3]  S. Antai,et al.  Response surface modeling and optimization of major medium variables for glycolipopeptide production , 2017 .

[4]  M. Sohaib,et al.  Plant and bacterial proteases: A key towards improving meat tenderization, a mini review , 2016 .

[5]  E. Tambourgi,et al.  Azocasein Substrate for Determination of Proteolytic Activity: Reexamining a Traditional Method Using Bromelain Samples , 2016, BioMed research international.

[6]  T. Nwagu,et al.  Production of a thermostable alkaline protease from alkalophilic Kocuria varians grown on various agricultural wastes , 2015 .

[7]  Padma Singh,et al.  Isolation and characterization of protease producing Bacillus sp from soil , 2015 .

[8]  Saraswathy Nagendran,et al.  PROTEASE: AN ENZYME WITH MULTIPLE INDUSTRIAL APPLICATIONS , 2014 .

[9]  S. Pradeep,et al.  Versatility of microbial proteases , 2013 .

[10]  V. N. Jeyadharshan.  Production and Partial Purification of Protease by Actinomyces Species , 2013 .

[11]  J. V. van Dijl,et al.  Membrane Proteases in the Bacterial Protein Secretion and Quality Control Pathway , 2012, Microbiology and Molecular Reviews.

[12]  M. Basri,et al.  Optimization of physical factors affecting the production of thermo-stable organic solvent-tolerant protease from a newly isolated halo tolerant Bacillus subtilis strain Rand , 2009, Microbial cell factories.

[13]  P. Vaithanomsat.  Silk Degumming Solution as Substrate for Microbial Protease Production , 2008 .

[14]  Fuensanta Máximo,et al.  Utilization of response surface methodology to optimize the culture media for the production of rhamnolipids by Pseudomonas aeruginosa AT10 , 2002 .

[15]  Q. Beg,et al.  Bacterial alkaline proteases: molecular approaches and industrial applications , 2002, Applied Microbiology and Biotechnology.

[16]  Ph.D. Christopher W. Wharton B.Sc.,et al.  Molecular Enzymology , 1981, Tertiary Level Biology.

[17]  S. T. Cowan.  Bergey's Manual of Determinative Bacteriology , 1948, Nature.