A Single-Step Purification of Cauliflower Lysozyme and Its Dual Role Against Bacterial and Fungal Plant Pathogens

Abstract

A novel lysozyme from cauliflower was purified in a single step, for the first time, using Sephadex G100 column chromatography. The purified lysozyme exhibited a homogenized single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and its molecular mass was calculated to be 22.0 kDa. The purified lysozyme showed activity between 30 to 60 °C with 40 °C as the optimum temperature for its maximal activity. Although the purified lysozyme was functional at pH ranges between 3.0 and 9.0, the optimum pH for the enzyme activity was 8.0. By Michaelis-Menten equation, the threshold substrate concentration for the optimal enzyme activity was calculated to be 133.0 μg. The purified lysozyme showed extraordinary activity against plant pathogenic bacteria and fungi. At 10-μg concentrations, it inhibited the growth of plant pathogenic bacteria such as Pseudomonas syringae, Xanthomonas campestris, and Erwinia carotovora exhibiting 4.28, 5.90, and 3.88-fold inhibition, respectively. Further, it also completely inhibited the conidial germination of Archemonium obclavatum and, to a very large extent, other fungal species such as Fusarium solani (79.3 %), Leptosphaeria maculans (88.6 %), Botrytis cinera (73.3 %), Curvularia lunata (68 %), Rhizoctonia solani (79.6 %), and Alternaria alternata (83.6 %).

Recent Insights Into the Prognostic and Therapeutic Applications of Lysozymes

“…Lysozyme isolated and purified from cauliflower tissue has shown the antibacterial and antifungal capability, which can affect the growth of plant pathogenic fungi and bacteria (Manikandan et al, 2015). Galleria mellonella lysozyme can resist the fungus Candida albicans (Sowa-Jasilek et al, 2016).…”

“…Hydrolyzes cell wall of Gram-positive bacteria (enzyme activity) (Ibrahim et al, 1991;Ibrahim et al 1992;Ibrahim et al 1994;Pellegrini et al 2000;Mine et al 2004;Derde et al 2013;Zhang et al 2016) Insert into and form pores in negatively charged bacterial membranes Antifungal Intact and/or peptides Enzymatic activity (Fiolka et al 2005;Wang et al 2005;Manikandan et al 2015;Hernandez-Tellez et al 2017;Sebaa et al 2017) Cationic nature leading to membrane destabilization Agglutination effect Immune modulator…”

“…For example, lysozyme isolated from Pithecellobium dulce seeds has antifungal activity against Macrophomina phaseolina ( Gálvez-Iriqui et al, 2020 ). Lysozyme isolated and purified from cauliflower tissue has shown the antibacterial and antifungal capability, which can affect the growth of plant pathogenic fungi and bacteria ( Manikandan et al, 2015 ). Galleria mellonella lysozyme can resist the fungus Candida albicans ( Sowa-Jasilek et al, 2016 ).…”

“…The lysozyme purified from mung bean further confirmed this result ( Wang et al, 2005 ). Other studies have shown that the antifungal activity of human lysozyme against common bacteria in patients with chronic rhinosinusitis is more than 80%, including Aspergillus fumigatus , Penicillium sp., Acremonium sp., Candida parasilopsis , and C. albicans ( Manikandan et al, 2015 ). Chitosan and lysozyme coating had a good preservation effect on the quality of large yellow croaker ( Larimichthys crocea ) during cold storage with no toxic and side effects ( Wu et al, 2018 ).…”

A Solid-State Fermentation (SSF) supplement improved performance, digestive function and gut ultrastructure of rainbow trout (Oncorhynchus mykiss) fed plant protein diets containing yellow lupin meal

“…Residual A. niger from the SSF procedure will inevitably be present within the diet, so theoretically lysozyme activity could respond to the presence of these fungal polysaccharides, regardless of their non-pathogenic threat. Indeed, lysozyme derived from insects and plants alike is believed to play a pivotal role in defence against common fungal infections (Javar et al, 2015;Manikandan et al, 2015) and its use in fungicidal drugs is of interest in human pharmacology (Woods et al, 2012). Furthermore, a recent study which supplemented polysaccharides from the mycelia of caterpillar fungus (Cordyceps sinensis), into the diets of white shrimp (Litopenaeus vannamei), stimulated a prolific increase in haemato-lysozyme activity (Deng et al, 2015).…”