Immobilization and characterization of lipase loaded on fe3o4 nanoparticles and produced from haloalkalophilis kocuria polaris-wrs3

A Gram-positive, cocci, thermoduric, haloalkalotolerant, and lipolytic bacterium designated strainWRS3, was isolated from salty soda soil sample collected from Wadi El-Ryan, Egypt. It is extremetolerant , pH10 and NaCl 22%. This isolate was identified as avariety of Kocuria polria.Strain WRS3 grows at different ranges of temperatures 15-50°C, pH 7.0-12.5 and salinity 3.0–30% NaCl (w/v).Euclidean distance was 97% between isolate WRS3 variety under Kocuriapolria.Over all phenetic relationship of isolate WRS3 was considered as Gram positive Phylum,variety of halo alkalo tolerant thermoduric of Kocuria, domain Eubacteria. Olive oil was the most suitable substrate for maximum lipase production by Kocuria polria-WRS3.Iron oxide (Fe3O4) magnetic nanoparticles and magnetite silica-coated (MSI) nanoparticles were used as immobilization material.Lipase was covalently bound to the silica-coated -functionalized magnetic nanoparticles with excellent properties have been successfully prepared using the chemical co-precipitation. The transmission electron microscope images revealed the size of the uncoated magnetite particles to be 200–500 nm and those of the coated particles about 300 nm. The best environmental conditions for lipase production by Kocuri apolria-WRS3 isolate indicated that the pH and temperature of the immobilized lipase were 8-9.5 and 40°C, respectively and, salt concentration range from 10% to 18% NaCl.Also,high enzymes productivity was obtained at aeration speed 150- 220 rpm between 48h. to 72h. Different types of oils (olive oil, corn oil, sun flower oil, punt oil and cotton oil)were tested for the selection of the best substrate for free lipase. Extracellular lipase produced was stimulated and exhibited the highest productivity bythe addition of peptone + histidine or casoamino acid + histidine to the product media. Highly lipase from Kocuri apolria-WRS3 exhibited better resistance to commercial detergents.Lipase activity was strongly resistance to Sodium Dodecyl Sulphate (SDS)and Triton X-100Arial, Tween 80 and Persil, respectively.The remarkable productivity of Kocuriapolria-WRS3 lipase in this range has proved to be a potential alkaline-saline lipase.

Author: 
Rawhia Arafa
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