DOI: https://doi.org/10.14710/jksa.23.11.383-389

Freeze-thaw system for thermostable β-Galactosidase isolation from Gedong Songo Geobacillus sp. isolate

1Chemical Engineering Department, Faculty of Engineering, Universitas Serang Raya, Indonesia

2Chemistry Department, Faculty of Sciences and Mathematics, Diponegoro University, Indonesia

Received: 18 Aug 2020; Revised: 18 Nov 2020; Accepted: 20 Nov 2020; Published: 30 Nov 2020.
Open Access Copyright 2020 Jurnal Kimia Sains dan Aplikasi under http://creativecommons.org/licenses/by-sa/4.0.

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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.
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Keywords: freeze-thawing; acetone-dry ice; β-galactosidase; ONPG; Geobacillus sp
Funding: Kementerian Riset Teknologi Dan Pendidikan Tinggi Republik Indonesia

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Section: Research Articles
Language : EN
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