L-Glutaminase Synthesis by Marine Halomonas meridiana Isolated from the Red Sea and Its Efficiency against Colorectal Cancer Cell Lines

Abstract

L-glutaminase is an important anticancer agent that is used extensively worldwide by depriving cancer cells of L-glutamine. The marine bacterium, Halomonas meridian was isolated from the Red Sea and selected as the more active L-glutaminase-producing bacteria. L-glutaminase fermentation was optimized at 36 h, pH 8.0, 37 °C, and 3.0% NaCl, using glucose at 1.5% and soybean meal at 2%. The purified enzyme showed a specific activity of 36.08 U/mg, and the molecular weight was found to be 57 kDa by the SDS-PAGE analysis. The enzyme was highly active at pH 8.0 and 37 °C. The kinetics’ parameters of Km and Vmax were 12.2 × 10−6 M and 121.95 μmol/mL/min, respectively, which reflects a higher affinity for its substrate. The anticancer efficiency of the enzyme showed significant toxic activity toward colorectal adenocarcinoma cells; LS 174 T (IC50 7.0 μg/mL) and HCT 116 (IC50 13.2 μg/mL). A higher incidence of cell death was observed with early apoptosis in HCT 116 than in LS 174 T, whereas late apoptosis was observed in LS 174 T more than in HCT 116. Also, the L-glutaminase induction nuclear fragmentation in HCT 116 was more than that in the LS 174T cells. This is the first report on Halomonas meridiana as an L-glutaminase producer that is used as an anti-colorectal cancer agent.

In Vitro Cytotoxicity Efficiency of Extracellular L-Glutaminase Produce by Mesophilic Bacteria, Kurthia Gibsonii on Lncap, MDA-MB 231 and Hepg-2 Cell Lines

“…Thus, marine bacteria have recently attracted attention for the Lglutaminase production (33). Halomonas meridian was first reported as an L-glutaminase producer that is used as an anti-colon cancer agent (34). In the present study, the produced L-glutaminase by the isolated soil bacterial strain Kurthia gibsonii showed potential anticancer activity against the tested cell lines with IC50 values of 30.11, 12.09, and 18.15 μg/ ml for LNCaP, HepG-2 and MDA-MB 231, respectively.…”

Natural Marine Products: Anti-Colorectal Cancer In Vitro and In Vivo

“…P6 showed a profound anti-tumor effect and induced tumor cell apoptosis by activating the p38 MAPK signaling pathway, promoting ROS production and intracellular Ca 2+ overload in CRC cells [22]. L-glutaminase was shown to drive early and late apoptosis in LS-174-T and HCT-116 tumor cells, as demonstrated by the acridine orange/ethidium bromide (AO/EtBr) double staining assay [17]. Ohmyungsamycin A, a novel cyclic peptide identified from a marine Streptomyces sp., inhibited the proliferation and tumor growth of HCT-116 human CRC cells and induced apoptosis also associated with the modulation of caspase family proteins, in addition to inducing G0/G1 phase cell cycle arrest [18].…”

“…Peptides are amphoteric compounds containing carboxyl and amino groups, which are dehydrated from amino acids, and are commonly found in marine organisms, such as bacteria, tunicates, mollusks, and sponges, among others. For example, L-glutaminase, ohmyungsamycin A, actinomycin V, androsamide, laxaphycin B4, and laxaphycin A2 (Table 2) are of bacterial origin [17][18][19][20][21], the FDA-approved dipeptide plitidepsin is of tunicates origin, and the peptide P6 (Table 2) is of mollusk origin [5,22]. Peptides can be divided into short linear peptides, long linear peptides, cyclic peptides, bicyclic peptides, thiopeptides, tetrapeptides, and so on, based on their chemical structure.…”

Enhancement of Halophilic Glutaminase Producing by Tetragenococcus muriaticus FF5302 in Bioreactor

“…Glutaminases are produced by a variety of microorganisms, including Aeromonas veronii [5], Alcaligenes faecalis [6], Aspergillus flavus [7], A. niger [8], A. oryzae [9], Bacillus subtilis [10], B. pasteurii Trends Sci. 2023; 20(4): 6504 2 of 11 [11], Clostridium welchii [12], Escherichia coli [13], Halomonas meridiana [14], Micrococcus luteus [15], Lactobacillus rhamnosus [16], Pseudomonas stutzeri [17], Rhizobium etli [18], Streptomyces canarius [19], Vibrio costicola [20], and Zygosaccharomyces rouxii [6], were isolated and well-studied. As expected, the glutaminases of these microorganisms differ in their biochemical and physicochemical properties Microorganisms are regarded as an essential source of glutaminase since the vast majority of glutaminases sold in commercial settings are derived from bacteria.…”