Cloning and Expression of the Chitinase Encoded by ChiKJ406136 from Streptomyces Sampsonii (Millard & Burr) Waksman KJ40 and Its Antifungal Effect

Abstract

The present study demonstrated that the chitinase gene ChiKJ406136 of Streptomyces sampsonii (Millard & Burr) Waksman KJ40 could be cloned using a PCR protocol and expressed in Escherichia coli (Migula) Castellani & Chalmers BL21 (DE3), and the recombinant protein had antifungal effect on four forest pathogens (Cylindrocladium scoparium Morgan, Cryphonectria parasitica (Murrill) Barr, Neofusicoccum parvum Crous, and Fusarium oxysporum Schl.) and also had the biological control effects on Eucalyptus robusta Smith leaf blight, Castanea mollissima BL. blight, Juglans regia L. blight and J. regia root rot. The results showed that ChiKJ406136 was efficiently expressed and a 48 kilodalton (kDa) recombinant protein was obtained. No significant change in protein production was observed in the presence of different concentrations of IPTG (isopropyl-b-D-thio-galactoside). The purified protein yield was greatest in the 150 mmol/L imidazole elution fraction, and the chitinase activities of the crude protein and purified protein solutions were 0.045 and 0.033 U/mL, respectively. The antifungal effects indicated that mycelial cells of the four fungi were disrupted, and the control effects of the chitinase on four forest diseases showed significant differences among the undiluted 10-and 20-fold dilutions and the control. The undiluted solution exhibited best effect. The results of this study provide a foundation for the use of S. sampsonii as a biocontrol agent and provides a new source for the chitinase gene, providing a theoretical basis for its application. & Grigoraki, Trichophyton sp. (Castell.) Sabour. [18] and Rhizoctonia violacea (Tul.) Pat. [20]. Previous studies demonstrated that the bioactive compounds of S. sampsonii have important applications in various fields [3]. For example, crude extracts showed antitumor activity against glioblastoma multiforme (GBM) cells, inhibiting cell growth by 70.04% [25], and the supernatant of a S. sampsonii culture showed biological activity against the root-knot nematode [26]. In the purified components, soil isolates of S. sampsonii can produce heptaene polyene antibiotics [18,27], In addition, S. sampsonii has been shown to produce hydrolytic enzymes, such as amylase, chitinase, protease, and lipase [19]. Studies at the molecular level have focused on strain identification and the phylogenesis of related species [28][29][30][31][32][33][34][35]. The complete genome sequence of Streptomyces sampsonii KJ40 was recently described by our lab [36], resulting in the discovery of a large number of gene encoding chitinases and enzymes involved in secondary metabolite production. However, little is known regarding the metabolic pathways and genetic regulation in this strain, limiting its practical application.Chitin, a linear polymer of β-1,4-glucosidicosamine (GlcNAC), is the second most abundant polysaccharide in nature. Chitin can be degraded by chitinolytic enzymes, that is chitinase. Chitinases (EC 3.2.1.14) are widely present in a great variety of organisms, including insects, fungi,...

Comparative Genomics of Marine Sponge-Derived Streptomyces spp. Isolates SM17 and SM18 With Their Closest Terrestrial Relatives Provides Novel Insights Into Environmental Niche Adaptations and Secondary Metabolite Biosynthesis Potential

“…In the previously identified SM17 clade (Figure 2), in addition to its closely related terrestrial type-strain J1074, the clade also included the environmental isolates Streptomyces albidoflavus SM254, which was isolated from copper-rich subsurface fluids within an iron mine (Badalamenti et al, 2016); Streptomyces sampsonii KJ40, which was isolated from rhizosphere soil in a poplar plantation (Li et al, 2018); Streptomyces koyangensis VK-A60T, which was isolated from rhizosphere soil in a radish plantation (Lee et al, 2005); and Streptomyces sp. CLI2509, which is a fungus-derived isolate (Wyche et al, 2017).…”

Characterization and potential antifungal activities of three Streptomyces spp. as biocontrol agents against Sclerotinia sclerotiorum (Lib.) de Bary infecting green bean

“…Also, S. rochei secreted antimicrobial compounds able to inhibit the growth of all common microbial indicators (Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, and Saccharomyces cerevisiae). Additionally, Li et al (2018) stated that the antifungal activity of S. sampsonii crude extract inhibited the hyphal growth and destroyed fungal cell walls of 4 different pathogenic fungi, i.e., Cylindrocladium scoparium, Cryphonectria parasitica, Neofusicoccum parvum, and Fusarium oxysporum). Nevertheless, many researchers recorded that the Actinobacteria proved to be great importance in the field of biotechnology, as producers of benefits and bioactive secondary metabolites, i.e., antifungal, insecticidal, antibacterial, enzymes such as chitinase, antibiotics, and antihelminthic activities with extensive industrial, medical use, and as a tool for eco-friendly management in agricultural applications of plant pathogens (Katarzyna et al 2018;Salwan and Sharma 2020;and Sharma et al 2020).…”

Genome Mining Coupled with OSMAC-Based Cultivation Reveal Differential Production of Surugamide A by the Marine Sponge Isolate Streptomyces sp. SM17 When Compared to Its Terrestrial Relative S. albidoflavus J1074

“…The Streptomyces albidoflavus strain J1074 stems from the soil isolate Streptomyces albus G [23,24]. The Streptomyces sampsonii strain KJ40 was isolated from rhizosphere soil in a poplar plantation [53]. The Streptomyces sp.…”

“…The Streptomyces sp. FR-008 isolate has been shown to produce the antimicrobial compound FR-008/candicidin [61,62]; while the Streptomyces sampsonii KJ40 isolate has been shown to produce a chitinase that possesses anti-fungal activity against plant pathogens [53]. On the other hand, although no bioactive compounds have been characterised from Streptomyces albidoflavus SM254, this isolate has been shown to possess anti-fungal activity, specifically against the fungal bat pathogen Pseudogymnoascus destructans , which is responsible for the White-nose Syndrome [55,63].…”