Kinetic properties of Streptomyces canarius L- Glutaminase and its anticancer efficiency

Abstract

L-glutaminase was produced by Streptomyces canarius FR (KC460654) with an apparent molecular mass of 44 kDa. It has 17.9 purification fold with a final specific activity 132.2 U/mg proteins and 28% yield recovery. The purified L-glutaminase showed a maximal activity against L-glutamine when incubated at pH 8.0 at 40 °C for 30 min. It maintained its stability at wide range of pH from 5.0 11.0 and thermal stable up to 60 °C with Tm value 57.5 °C. It has high affinity and catalytic activity for L-glutamine (Km 0.129 mM, Vmax 2.02 U/mg/min), followed by L-asparagine and L-aspartic acid. In vivo, L-glutaminase showed no observed changes in liver; kidney functions; hematological parameters and slight effect on RBCs and level of platelets after 10 days of rabbit's injection. The anticancer activity of L-glutaminase was also tested against five types of human cancer cell lines using MTT assay in vitro. L-glutaminase has a significant efficiency against Hep-G2 cell (IC50, 6.8 μg/mL) and HeLa cells (IC50, 8.3 μg/mL), while the growth of MCF-7 cells was not affected. L-glutaminase has a moderate cytotoxic effect against HCT-116 cell (IC50, 64.7 μg/mL) and RAW 264.7 cell (IC50, 59.3 μg/mL).

Purification, characterization and anticancer efficiency of L-glutaminase from <i>Aspergillus flavus</i>

“…The high affinity of A. flavus glutaminase to L-glutamine as a substrate was recorded previously by Elshafei et al (2014) and Reda (2015) for Penicillium brevicompactum and Streptomyces canaries, respectively. While the low activity of L-glutaminase towards L-albizziin, and the absence of activity towards glutathione and L-γ-Glu-p-NA are in agreement with that recorded for L-glutaminase from Saccharomyces cerevisia and Debaryomyces spp.…”

“…Our results were in complete agreement with that recorded for the enzyme of Debaryomyces sp. (Dura et al, 2002), Streptomyces canaarius (Reda, 2015) and Aspergillus oryzae (Koibuchi et al, 2000). A higher optimum temperature was recorded for purified Lglutaminase from Aspergillus sojae (Ito et al, 2012).…”

“…The purified L-glutaminase of A. flavus was found to have a good activity at an alkaline value (pH 8) and this optimum pH is equal to that reported for purified L-glutaminase from Debaryomyces sp. (Dura et al, 2002 ), Streptomyces canaarius (Reda, 2015) and Penicillium politans NRC (Ali et al, 2009).…”

Production, purification and characterization of L-Glutaminase from Streptomyces sp. isolated from soil

“…The glutaminase purified from Micrococcus was also determined to be 49.9 kDa (Masuo et al, 2005). Reda FM (2015) reported production of L-glutaminase from Streptomyces canarius strain FR having molecular mass of 44 kDa and showed maximum enzyme activity at pH 8.0 at 40 o C incubated for 30 min. The enzyme showed good anticancer activity against five human cancer cell lines when tested using MTT in-vitro assay.…”

Recombinantl-glutaminase obtained fromGeobacillus thermodenitrificansDSM-465: characterization andin silicoelucidation of conserved structural domains

“…5,6 Thus, Lglutaminase is an attractive target to block the energy route for the proliferation of glutamine-dependent tumour cells and responsible for their selective death. 7,8 Several other applications of the enzyme have been reported due to its effective antiretroviral activity; 9 moreover, the enzyme is used to improve the taste and aroma of the food products. 10 L-Glutaminase also has a signicant application in biosensors where it is used to monitor the level of glutamine in mammalian and hybridoma cells.…”