Separation, determination and antifungal activity test of the products from a new Bacillus amyloliquefaciens

Abstract

A new Bacillus amyloliquefaciens named ZJU-2011 was discovered, and the culture supernatant showed a strong inhibitory effect against Candida albicans. In this study, a novel method was developed to purify the antifungal compounds in high purity. The obtained products were analysed by high performance liquid chromatography and proven to be of high purity. Mass spectrometry showed that the molecular weights of the two bioactive components were 270 and 288, respectively, and their structures were determined to be bacilysin and chlorotetaine by using (1)H and (13)C nuclear magnetic resonance spectroscopy. To the best of our knowledge, this is the first time that B. amyloliquefaciens has been reported to produce bacilysin and chlorotetaine simultaneously. The minimum inhibitory concentration of chlorotetaine against six common fungal pathogens were determined to be in the range of 1.8-7.8 μg/mL.

Potential of a new strain of Bacillus amyloliquefaciens BUZ-14 as a biocontrol agent of postharvest fruit diseases

“…Some strains have been employed to develop commercial products such as Subtilex ® (B. subtilis MBI 600; BASF), registered in the USA by the EPA and used for the control of fruit, herb and vegetable diseases, amyloliquefaciens, closely related to B. subtilis, dedicates 8.5 % of its genome to produce several bioactive compounds with high antifungal activity such as lipopeptides including surfactin (Ahimou et al, 2000;Hsieh et al, 2004) iturin (Hsieh et al, 2008; A C C E P T E D M A N U S C R I P T Yu et al, 2002) or fengycin (Lin et al, 1999;Ongena et al, 2007) and several polyketides such as macrolactin, bacillaene, bacilysin, bacillomycin and difficidin (Chen et al, 2009). Furthermore, other antibiotics such as plantazolicin or chlorotetaine, all of which have substantial antimicrobial and antifungal activity, are also secreted (Scholz et al, 2011;Mudgal et al, 2013;Wang et al, 2016). Recently, Taegro® (B. amyloliquefaciens FZB24); Novozymes (Copenhagen, Denmark) and Syngenta (Basel, Switzerland) (Lecomte et al, 2016), recommended for suppressing selected soil-borne and foliar diseases of fruits and leafy vegetables, cucurbits, and ornamentals, have been registered in the USA by the EPA.…”

“…Although several authors have revealed the major role of fengycnin-like lipopeptides in the biological control of Bacillus species against fungal pathogens (Ongena et al, 2005;Romero et al, 2006;Hu et al, 2007;Alvarez et al, 2012) including M. fructicola (Lui et al, 2011;Yánez-Mendizábal et al, 2012), the activity of other lipopeptides such as iturins (Yu et al, 2002;Arrebola et al, 2010) or other antimicrobial compounds such as plantazolicin (Scholz et al, 2010) or chlorotetaine (Wang et al, 2016) and the presence of synergistic effects (Maget-Dana et al, 1992;Lui et al, 2011;Tao et al, 2011) could not be ruled out. Thus, the characterization of the bioactive compounds produced by BUZ-14 in terms of identity, antifungal activity using non-producing mutants, total and individual amounts, relative proportions and progressive accumulation both in culture media and in fruit are key aspects to understand the mechanism of action of this BCA against both fungal pathogens.…”

Antimicrobial and mechanism of antagonistic activity of Bacillus sp. A2 against pathogenic fungus and bacteria: The implication on honey's regulatory mechanism on host's microbiota

“…The experiments were done three independent times, with three replicates each. A p-value < .05 was considered to be significant, * denoted p < .05, ** denoted p < .01, and *** denoted p < .001 & Liu, 2017), honey (Zhao, de Jong, Zhou, & Kuipers, 2015), soil, rhizosphere (Lim et al, 2016), sea (Wang, Wu, Chen, Lin, & Yang, 2016), as an endophyte (White et al, 2014). B. amyloliquefaciens has been investigated for a variety of its beneficial properties, among them remediation of aquatic water; as biofertilizer (Chowdhury, Hartmann, Gao, & Borriss, 2015;Pretorius, van Rooyen, & Clarke, 2015;White et al, 2014), removal of mycotoxin from animal feeds (Chang, Wu, Wu, Dai, & Sun, 2015;A.…”

“…albicans (Ndlovu, Rautenbach, Vosloo, Khan, & Khan, 2017;Wang et al, 2016), Clostridium difficille (Geeraerts et al, 2015), and Listeria monocytogenes (Lim et al, 2016); industrially for production of protease and amylase enzyme and as source of antimicrobials, for example, macrolactin A and E, bacillomycin D, (Yuan et al, 2014). The origin of Bacillus sp.…”

Antimicrobial Bacillus: Metabolites and Their Mode of Action

“…Glucosamine-6-phosphate synthetase (G6PS) is an enzyme that catalyzes the production of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), which is a precursor for peptidoglycan synthesis [ 18 ]. Bacilysin (2) and its chlorinated derivative chlorotetaine (3) ( Figure 3 ) were first isolated from B. subtilis A14 and B. amyloliquefaciens ZJU-2011, respectively [ 19 , 20 ]. Both compounds are active against a broad range of bacteria, with bacilysin inhibiting E. coli at MIC = 0.001 µg/mL and chlorotetaine inhibiting Candidas spp.…”