ABSTRACT
Surfactants are used to prevent the irreversible aggregation of partially refolded proteins and they also assist in protein refolding. We have reported the design and screening of gemini surfactant to stabilize bovine serum albumin (BSA) with the help of computational tool (iGEMDOCK). A series of gemini surfactant has been designed based on bis-N-alkyl nicotinate dianion via varying the alkyl group and anion. On changing the alkyl group and anion of the surfactant, the value of Log P changes means polarity of surfactant can be tuned. Further, the virtual screening of the gemini surfactant has been carried out based on generic evolutionary method. Herein, thermodynamic data was studied to determine the potential of gemini surfactant as BSA stabilizer. Computational tools help to find out the efficient gemini surfactant to stabilize the BSA rather than to use the surfactant randomly and directionless for the stabilization. It can be confirmed through the experimental techniques. Previously, researcher synthesized one of the designed and used gemini surfactant to stabilize the BSA and their interactions were confirmed through various techniques and computational docking. But herein, the authors find the most competent gemini surfactant to stabilize BSA using computational tools on the basis of energy score. Different from the single chain surfactant, the gemini surfactants exhibit much stronger electrostatic and hydrophobic interactions with the protein and are thus effective at much lower concentrations. Based on the present study, it is expected that gemini surfactants may prove useful in the protein stabilization operations and may thus be effectively employed to circumvent the problem of misfolding and aggregation.
Subject(s)
Niacin/analogs & derivatives , Niacin/chemistry , Serum Albumin, Bovine/chemistry , Surface-Active Agents/chemistry , Animals , Anions , Cattle , Hydrogen Bonding , Hydrophobic and Hydrophilic Interactions , Models, Chemical , Models, Molecular , Protein Conformation , Structure-Activity RelationshipABSTRACT
A protease-producing, crude oil degrading marine isolate was identified as Nocardiopsis sp. on the basis of the morphology, cell wall composition, mycolic acid analysis and DNA base composition. The Nocardiopsis produces two extracellular proteases, both of which are alkaline serine endopeptidases. Protease I was purified to homogeneity by chromatography on CM-Sephadex at pH 5.0 and pH 9.0. Protease II was purified using DEAE-cellulose, Sephadex G-50, phenyl-Sepharose and hydroxyapatite chromatography. Protease I and II had almost similar M(r) of 21 kDa (Protease I) and 23 kDa (Protease II), pI of 8.3 and 7.0 respectively with pH and temperature optima for activity between 10.0 and 11.0 and about 60 degrees C. Specific activities were 152 and 14 U/mg respectively on casein. However, Protease I was antigenically unrelated to Protease II. Both proteases were endopeptidases and required extended substrate binding for catalysis. Both proteases had collagenolytic and fibrinolytic activity but only Protease I had elastinolytic activity. The proteases were chymotrypsin-like with respect to their amino acid compositions and N-terminal sequences.
Subject(s)
Actinomycetales/enzymology , Serine Endopeptidases/metabolism , Actinomycetales/classification , Actinomycetales/isolation & purification , Amino Acid Sequence , Chromatography, Gel , Chromatography, Ion Exchange , Kinetics , Molecular Sequence Data , Seawater/microbiology , Sequence Alignment , Sequence Homology, Amino Acid , Serine Endopeptidases/chemistry , Serine Endopeptidases/isolation & purification , Substrate Specificity , ThermodynamicsABSTRACT
An actinomycete isolated from an oil-contaminated marine environment and identified as Nocardiopsis sp. degraded hydrocarbons and also produced extracellular protease. Conditions for crude oil degradation and simultaneous production of extracellular protease were studied. An alternative approach for bio-augmented clean-up of oil spills using a micro-organism capable of degrading hydrocarbons and recruiting organic nitrogen by producing proteases is reported.
