Fe(III)-based immobilized metal–affinity chromatography (IMAC) method for the separation of the catechol siderophore from Bacillus tequilensis CD36

Abstract

Catechol siderophore plays an important role in microbial ecology, agriculture, and medicine, but its research is often limited by the difficulty in acquisition of it in large quantities. Based on evidence from the coordination chemistry and chemical biology, catechol siderophore could chelate Fe with high affinity. Therefore, Fe(III)-based immobilized metal-affinity chromatography (IMAC) was applied to capture siderophore from the culture filtrate of CD36. The ethanol-precipitated sample and the separated sample from Fe(III)-based IMAC were analyzed by liquid chromatography-mass spectrometry. According to the result, the pure siderophore DHB-Gly-Thr could be extracted from the ethanol-precipitated sample. Compared with other purifications, Fe(III)-based IMAC was convenient and had fewer steps. In addition, it also reduced the use of toxic chemical solvents in some traditional extraction process, such as extraction and ion exchange chromatography. Fe(III)-based IMAC was successfully used in separation of the catechol siderophore from CD36. The results revealed that Fe(III)-based IMAC was an efficient and environmentally friendly method for the separation and purification of catechol siderophore.

Genome mining and UHPLC–QTOF–MS/MS to identify the potential antimicrobial compounds and determine the specificity of biosynthetic gene clusters in Bacillus subtilis NCD-2

“…Microorganisms and culture conditions B. subtilis NCD-2 was routinely grown at 37 °C on Luria Bertani medium. For lipopeptide, bacillaene, bacilysin, bacillibactin and subtilosin production, strain NCD-2 was grown in Landy broth [58], PA medium [59], MSA medium [60], and TSB medium [61] at 30 ℃ and 180 rpm. The phytopathogen Botrytis cinerea BC-10 was used for antifungal activity test following the method described by Guo et al [28] with some modi cations.…”

“…For bacilysin, strain NCD-2 was cultured in 100 mL PA medium at 30℃ for 72 h with shaking at 180 rpm, and bacilysin was extracted with ice-cold ethanol as described by Wu et al [59]. For bacillibactin, strain NCD-2 was cultured in 100 mL MSA medium at 30℃ for 72 h, and bacillibactin was extracted with ethanol as described by Li et al [60]. For subtilosin, strain NCD-2 was cultured in 100 mL TSB medium at 30℃ for 72 h, and subtilosin was extracted with precipitation with 65% ammonium sulphate as described by Charles et al [61].…”

Genome mining and UHPLC–QTOF–MS/MS illuminate the potential antimicrobial active compounds and specificity of biosynthetic gene clusters in Bacillus subtilis NCD-2

“…Microorganisms and culture conditions B. subtilis NCD-2 was routinely grown at 37 °C on Luria Bertani medium. For lipopeptide, bacillaene, bacilysin, bacillibactin and subtilosin production, strain NCD-2 was grown in Landy broth [63], PA medium [64], MSA medium [65], and TSB medium [66] at 30 ℃ and 180 rpm. Phytopathogen Botrytis cinerea BC-10 was used for antifungal activity test following the method described by Guo et al [29] with some modi cations.…”

“…For bacilysin, strain NCD-2 was cultured in 100 mL PA medium at 30℃ for 72 h with shaking at 180 rpm, and the bacilysin was extracted by ice-cold ethanol as described by Wu et al [64]. For bacillibactin, strain NCD-2 was cultured in 100 mL MSA medium at 30℃ for 72 h, and the bacillibactin was extracted by ethanol as described by Li et al [65]. For subtilosin, strain NCD-2 was cultured in 100 mL TSB medium at 30℃ for 72 h, and the subtilosin was extracted by precipitation with 65% ammonium sulphate as described by Charles et al [66].…”

Evaluation of the Strain Bacillus amyloliquefaciens YP6 in Phoxim Degradation via Transcriptomic Data and Product Analysis

“…The extracts containing phoxim were analyzed by HPLC-MS. The system, column, and method were the same as those used in Li et al [47]. The different conditions were as follows: mobile phase A (methanol), mobile phase B (H 2 O, 0.1% formic acid), 0.3 mL/min of flow rate, 1 µL of sample size, a source temperature of 100 °C, a desolvation temperature of 400 °C, capillary voltage of 3.5 kV, and cone voltage of 30 V.…”