Freeze-thaw system for thermostable β-Galactosidase isolation from Gedong Songo Geobacillus sp. isolate
| Autor: | Nies Suci Mulyani, Ken Ima Damayanti, Agustina L. N. Aminin |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Ammonium sulfate freeze-thawing 030306 microbiology Thermophile β-galactosidase Substrate (chemistry) Chemistry 03 medical and health sciences chemistry.chemical_compound Membrane Enzyme acetone-dry ice chemistry Lowry protein assay onpg geobacillus sp Specific activity Food science Lactose QD1-999 030304 developmental biology |
| Zdroj: | Jurnal Kimia Sains dan Aplikasi, Vol 23, Iss 11, Pp 383-389 (2020) |
| ISSN: | 2597-9914 1410-8917 |
| Popis: | The effective isolation of intracellular enzymes from thermophilic bacteria is challenging because of their sturdy membrane. On the other hand, the low-cost and nontoxic method is essential for industrial food enzymes. The freeze-thaw cycles using acetone-dry ice as a frozen system was studied for efficient isolation of thermostable b-galactosidase from Geobacillus sp. dYTae-14. This enzyme has been known for application in the dairy industry to reduce the lactose content. In this study, the freeze-thaw method was performed with cycle variations 3, 5, and 7 cycles. Acetone-dry ice (-78°C) is used as a frozen system and boiling water for thawing. The b-galactosidase activity was assayed using ortho-Nitrophenyl-β-galactoside (ONPG) as substrate and protein content determined with the Lowry method. The results show that the most effective freeze-thaw is five cycles. The enzyme’s highest specific activity is 3610.13 units/mg proteins at 40-60 % ammonium sulfate saturation, with a purity value of 2.52. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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