Abstract
Three-dimensional structure of thermostable lipase is much sought after nowadays as it is important for industrial application mainly found in the food, detergent, and pharmaceutical sectors. Crystallization utilizing the counter diffusion method in space was performed with the aim to obtain high resolution diffracting crystals with better internal order to improve the accuracy of the structure. Thermostable T1 lipase enzyme has been crystallized in laboratory on earth and also under microgravity condition aboard Progress spacecraft to the ISS in collaboration with JAXA (Japanese Aerospace Exploration Agency). This study is conducted with the aims of improving crystal packing and structure resolution. The diffraction data set for ground grown crystal was collected to 1.3 Å resolution and belonged to monoclinic C2 space group with unit cell parameters a = 117.40 Å, b = 80.95 Å, and c = 99.81 Å, whereas the diffraction data set for space grown crystal was collected to 1.1 Å resolution and belonged to monoclinic C2 space group with unit cell parameters a = 117.31 Å, b = 80.85 Å, and c = 99.81 Å. The major difference between the two crystal growth systems is the lack of convection and sedimentation in microgravity environment resulted in the growth of much higher quality crystals of T1 lipase.
Highlights
Lipases are enzymes which are present in many different organisms
Only two thermostable lipases were reported grown by counter diffusion method, which are L42 lipase and L2 lipase
The reflections of ground-grown crystal was indexed on a centred monoclinic lattice (C2 with unit cell parameters a = 117.40 A, b = 80.95 A, c = 99.81 A, α = 90∘, β = 96.76∘, and γ = 90∘)
Summary
Lipases (triacylglycerol acylhydrolase) are enzymes which are present in many different organisms. It catalyze both the hydrolysis of triglycerides and the synthesis of esters formed from alcohol and long chain fatty acids. Enzymes from thermophiles often show more stability towards organic solvents and exhibit higher activity at elevated temperatures. This thermostable enzyme became an important requirement as a biocatalyst in industry including detergent, food, pharmaceuticals, cosmetic, textiles, fine chemicals, and biodiesel [1]. To better understand the mechanism of the enzyme, it is important to elucidate the three-dimensional structure of the enzyme.
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