Purification of Geobacillus thermodenitrificans thermostable lipase / Nik Raikhan Nik Him and Muhammad Syafiq Abu Hassan
The demand for a structured method in producing high quality lipases has tremendously increased lipase purification technology within specific range of its original function. Crude lipase enzyme produced by a locally isolated Geobacillus thermodenitrificans (LGT) has demonstrated a notable potential...
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| Main Authors: | , |
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| Format: | Article |
| Language: | en |
| Published: |
Universiti Teknologi MARA
2024
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| Subjects: | |
| Online Access: | https://ir.uitm.edu.my/id/eprint/107380/1/107380.pdf https://ir.uitm.edu.my/id/eprint/107380/ |
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| Summary: | The demand for a structured method in producing high quality lipases has tremendously increased lipase purification technology within specific range of its original function. Crude lipase enzyme produced by a locally isolated Geobacillus thermodenitrificans (LGT) has demonstrated a notable potential for various bio-deinking capacities and other enzymatic actions based on the credential amount used. The enzyme production was set up in an airlift fermenter system using a cultivation medium prepared from (w/v) 1.25% glucose and yeast extract, 0.75% NaCl and 0.10% olive oil. Fermentation was accomplished in 24 hours with an air flow rate of 1.00 L/min. Physical parameters were maintained at 7.0% (v/v) inoculum size and pH 6.8. The extracted extracellular crude lipase was purified to homogeneity using four-step procedures. The acetone precipitation, Sephadex G-100 filtration chromatography and double steps of DEAE Sefarose CL-6B anion with exchange chromatography has resulted in a final yield of 25.00%. The molecular weight of the purified enzyme was estimated to be 33.5 kDa using an SDS-PAGE analysis. The lipase enzyme has a high potential to be further used for many industrial purposes and has been purified with 22.1-fold protein. The above functional reported characteristics of the LGT was considered a modest quality taking into consideration of its ability to stand high temperature range. |
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