Purification and Structural Analysis of the Effective Anti-TMV Compound ε-Poly-l-lysine Produced by Streptomyces ahygroscopicus

Abstract

Microbial secondary metabolites produced by actinomycetes are important natural products widely applied to control plant diseases. A variety of actinomycetes were isolated from soil samples collected from Tianzhu Mountain in Shenyang, China. A Streptomyces strain Shenyang Tianzhu (STZ) exhibits effective antiviral activity against Tobacco mosaic virus (TMV). The isolate was identified as Streptomyces ahygroscopicus based on its cultural, morphological, physiological, biochemical characteristics as well as the phylogenetic analysis using 16S rRNA sequences. To obtain the pure anti-TMV compound from Streptomyces STZ, the culture broth was subjected to Amberlite IRC-50 ion-exchange resin, SX-8 macroporous adsorption resin and Sephadex G-25 gel column chromatography. The purified active compound was confirmed to be ε-poly-l-lysine (ε-PL), with molecular mass in the range of 3454–4352 Da by structural analysis with infrared (IR), matrix-assisted laser desorption ionization-time-of-flight MS (MALDI-TOF), thin-layer chromatography (TLC) and high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR). The protective and curative effects of the purified compound ε-PL were tested and the results showed that the compound exhibited significant protective and curative activity against TMV. The potential application of ε-PL as an efficient anti-plant virus agent was expected.

Physiological and Proteomic Analysis of Penicillium digitatum in Response to X33 Antifungal Extract Treatment

“…Two grams of the fresh mycelium collected at 48 h was re-suspended in 50 mL PDB, which X33 antifungal extract was poured and adjusted to 0, 1.2, 2.4, and 4.8 mg/mL. The extracellular electric conductivity was measured at 0, 1, 2, 3, 4, and 5 h, and expressed as the amount of extracellular conductivity (μs/cm) ( Chen et al, 2019d ).…”

“…The MDA content was estimated by using thiobarbituric acid approach as described previously ( Zhiheng et al, 2014 ). The extraction and evaluation of H 2 O 2 in P. digitatum hyphae were conducted according to the method reported previously ( Chen et al, 2019d ). Frozen mycelia samples were homogenized in 5 mL of ice-cold acetone and centrifuged at 15,000 × g for 20 min.…”

“…The extracellular conductivity of P. digitatum was estimated according to previous study (Tao et al, 2019) with modulations on employing an electrical conductivity meter (model ST3100c/F, Ohaus Co., New Jersey, United States) (Chen et al, 2019d). Two grams of the fresh mycelium collected at 48 h was re-suspended in 50 mL PDB, which X33 antifungal extract was poured and adjusted to 0, 1.2, 2.4, and 4.8 mg/mL.…”

Screening of Marine Bioactive Antimicrobial Compounds for Plant Pathogens

“…During separation, organic compounds can pass through a certain solvent according to the adsorption force and molecular weight [ 79 ]. Through the macroporous adsorption resin adsorption method, Chen et al separated the anti-tobacco mosaic virus (TMV) ε-Poly-l-lysine compounds from Streptomyces ahygroscopicus [ 80 ], and He et al separated the anti-gray mold compound tetramycin A from ZC-G-5 [ 81 ].…”

Extraction and purification of ε-poly-l-lysine from fermentation broth using an ethanol/ammonium sulfate aqueous two-phase system combined with ultrafiltration

“…7 Chen et al employed an Amberlite IRC-50 ion-exchange resin, a SX-8 macroporous adsorption resin and Sephadex G-25 gel column chromatography to purify 3-PL from a culture of S. ahygroscopicus, and this was used for its chemical structure identication. 8 In previous studies, we developed a potential industrial scheme for 3-PL separation and purication from fermentation broth, including mycelia removal by occulation, protein removal by 30 kDa ultraltration, ion-exchange adsorption, decolorization with a macroporous resin and desalting by 1 kDa ultraltration. A 90.2% purity and 75% recovery were achieved.…”