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All Journal JURNAL KIMIA SAINS DAN APLIKASI
Ken Ima Damayanti
Chemical Engineering Department, Faculty of Engineering, Universitas Serang Raya Serang
Chemical Engineering, Chemistry & Bioengineering Chemistry Engineering

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Freeze-thaw system for thermostable β-Galactosidase isolation from Gedong Songo Geobacillus sp. isolate Ken Ima Damayanti; Nies Suci Mulyani; Agustina L. N. Aminin
Jurnal Kimia Sains dan Aplikasi Vol 23, No 11 (2020): Volume 23 Issue 11 Year 2020
Publisher : Chemistry Department, Faculty of Sciences and Mathematics, Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2994.949 KB) | DOI: 10.14710/jksa.23.11.383-389

Abstract

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.
Co-Authors Agustina L.N. Aminin Nies Suci Mulyani