Akademik Veri Yönetim Sistemi
Özdal M.(Yürütücü), Gülmez Ö., Gökçe E.
Yükseköğretim Kurumları Destekli Proje, 2019 - 2021
L-asparaginase (EC. 3.5.1.1, L-asparagine amidohydrolase), which is one of
the leading enzymes of industrial importance and used in cancer treatment,
has attracted great attention in the pharmacology sector in recent years.
In our study, different bacteria were isolated and purified from the
digestive systems of some animals and from the soil. By comparing the L-
asparaginase activities of the isolates with the zones they formed on the
M9 medium, bacteria were selected for the highest L-asparaginase production
potential. It was later determined whether these bacteria produce
glutaminase and urease. The isolates that produce glutaminase and L-
asparaginase that do not produce glutaminase and urease were determined by
sequencing Bacillus atrophaeus strain AspK1 (MW866485.1), Bacillus atrophaeus
strain AspK3 (MW872009.1) and Brevibacterium frigoritolerans strain AspS1
(MW866486.1) 16s rRNA. Among them, Bacillus atrophaeus strain AspK1 was found
to have the highest L-asparaginase activity (15.2 U/mL), and studies were
continued with this organism. The effects of carbon and nitrogen sources,
pH, temperature and incubation time on enzyme production were investigated.
L-asparaginase was first partially purified by ethanol precipitation, then
it could be purified by anion exchange chromatography. The purity of the
enzyme was checked by SDS-PAGE analysis and its molecular weight was
determined to be 42 kDa. The optimum temperature of the enzyme was found to
be 30 ℃, and the optimum pH to be 7. The effects of various substances on
L-asparaginase activity have been studied and it has been observed that
Mg 2+ , Ca 2+ and Mn 2+ ions increase the activity.