In Silico Designing of an Industrially Sustainable Carbonic Anhydrase Using Molecular Dynamics Simulation

Carbonic anhydrase (CA) is a family of metalloenzymes that has the potential to sequestrate carbon dioxide (CO₂) from the environment and reduce pollution. The goal of this study is to apply protein engineering to develop a modified CA enzyme that has both higher stability and activity and hence could be used for industrial purposes. In the current study, we have developed an in silico method to understand the molecular basis behind the stability of CA. We have performed comparative molecular dynamics simulation of two homologous α-CA, one of thermophilic origin (<i>Sulfurihydrogenibium</i> sp.) and its mesophilic counterpart (Neisseria gonorrhoeae), for 100 ns each at 300, 350, 400, and 500 K. Comparing the trajectories of two proteins using different stability-determining factors, we have designed a highly thermostable version of mesophilic α-CA by introducing three mutations (S44R, S139E, and K168R). The designed mutant α-CA maintains conformational stability at high temperatures. This study shows the potential to develop industrially stable variants of enzymes while maintaining high activity

Single Cell Oil from Oleaginous Yeast Grown on Sugarcane Bagasse-Derived Xylose: An Approach toward Novel Biolubricant for Low Friction and Wear

Yeast lipid as single cell oil (SCO) is evaluated as an alternative renewable source of vegetable oils for a biolubricant formulation. The <i>Rhodotorula mucilaginosa</i> IIPL32 yeast strain is cultivated on lignocellulosic pentosans derived from sugarcane bagasse to produce the SCO. The chemical composition and distribution of variable fatty acids in the yeast SCO are characterized by NMR, FTIR, and GC × GC analyses. The high viscosity index and a low pour point of yeast SCO owing to the favorable composition of saturated and unsaturated fatty acids promise its potential as a renewable and environmentally friendly lube base oil. The yeast SCO as lube base oil significantly reduced the coefficient of friction (72%) and wear (24%) compared to those of conventional mineral lube base oil (SN 150). The fatty acids in the yeast SCO formed a good quality tribo-chemical thin film on the engineering surfaces, which not only reduced the friction but also protected the contact interfaces against wear. This study demonstrates that yeast SCO being renewable, biodegradable, and nontoxic, provides favorable physicochemical and tribophysical properties for good quality lubricant formulation and it can be a good alternative to the conventional mineral lube oil-based lubricants