Home practices can mitigate furan and derivatives in vegetable-based infant meals.

This study assessed the impact of current home practices including reheating, standing, and stirring on mitigation of furan and its derivatives in vegetable-based infant meals. Three vegetable-based infant meals (vegetables alone, with fish, with meat) underwent different home practices including reheating, post-reheating standing (60, 120 and 240 s) and post-reheating stirring (30, 60, 120 and 240 s). Targeted quantification of furan, 2-methylfuran (2-MF) and 3-methylfuran (3-MF) and exploration of additional furan derivatives were undertaken in treated and untreated vegetable-based infant meals using SHS-GC-Q Exactive-Orbitrap-MS. For the three compounds, the quality of the measurements was first validated with suitable linearity, limits of quantification, precision and recoveries. A second step highlighted high concentrations of furan (78.5-103.9 µg/kg), 2-MF (4.8-10.1 µg/kg) and 3-MF (3.4-5.8 µg/kg) in the three vegetable-based infant meals before preparation and the assessment of the cumulative risk related to these three furan compounds confirmed the relevance of studying home mitigation strategies. The third step showed that post-reheating stirring was the most effective home practice for mitigation, with maximum observed reductions of 66.3, 59.9 and 57.7 % for furan, 2-MF and 3-MF, respectively. In a fourth step, a suspect screening approach carried out on SHS-GC-Q Exactive-Orbitrap MS data revealed the presence of 2-ethyl-, 2-ethyl-5-methyl-, 2-butyl- and 2-vinyl-furan in vegetable-based meals and showed a similar mitigation trend of home practices on the relative concentrations of these four additional furan derivatives. Finally, despite a significant mitigation reaching 69 % of the furan concentration, the combined effect of home practices on furan compounds was not sufficient to rule out the risk associated with the consumption of vegetable-based infant foods and additional options are discussed.

Quantitative interdependence of coeffectors, CcpA and cre in carbon catabolite regulation of Bacillus subtilis.

The phosphoproteins HPrSerP and CrhP are the main effectors for CcpA-mediated carbon catabolite regulation (CCR) in Bacillus subtilis. Complexes of CcpA with HPrSerP or CrhP regulate genes by binding to the catabolite responsive elements (cre). We present a quantitative analysis of HPrSerP and CrhP interaction with CcpA by surface plasmon resonance (SPR) revealing small and similar equilibrium constants of 4.8 +/- 0.4 microm for HPrSerP-CcpA and 19.1 +/- 2.5 microm for CrhP-CcpA complex dissociation. Forty millimolar fructose-1,6-bisphosphate (FBP) or glucose-6-phosphate (Glc6-P) increases the affinity of HPrSerP to CcpA at least twofold, but have no effect on CrhP-CcpA binding. Saturation of binding of CcpA to cre as studied by fluorescence and SPR is dependent on 50 microm of HPrSerP or > 200 microm CrhP. The rate constants of HPrSerP-CcpA-cre complex formation are k(a) = 3 +/- 1 x 10(6) m(-1).s(-1) and k(d) = 2.0 +/- 0.4 x 10(-3).s(-1), resulting in a K(D) of 0.6 +/- 0.3 nm. FBP and Glc6-P stimulate CcpA-HPrSerP but not CcpA-CrhP binding to cre. Maximal HPrSerP-CcpA-cre complex formation in the presence of 10 mm FBP requires about 10-fold less HPrSerP. These data suggest a specific role for FBP and Glc6-P in enhancing only HPrSerP-mediated CCR.

Cloning, sequencing, and characterization of the genetic region relevant to biosynthesis of the lipopeptides iturin A and surfactin in Bacillus subtilis.

Bacillus subtilis B3 was found to produce lipopeptides iturins and fengycin that have activity against several plant pathogens such as Fusarium graminearum, Rhizoctonia solani, Rhizoctonia cerealis, and Pyricularia grisea. A 3642-bp genomic region of B. subtilis B3 comprising srfDB3, aspB3, lpaB3, and yczEB3 genes that resulted in biosynthesis of surfactin in B. subtilis 168 was cloned, sequenced, and characterized. Among them, the srfDB3 gene encodes thioesterase, which is required for biosynthesis of surfactin in B. subtilis; the aspB3 gene encodes a putative aspartate aminotransferase-like protein; the lpaB3 encodes phosphopantetheinyl transferase, which shows high identity to the product of lpa-14 gene regulating the biosynthesis of iturin A and surfactin in B. subtilis RB14; the yczEB3 encodes a YczE-like protein with significant similarities in signal peptide and part of the ABC transport system. The genetic regions between the srfD gene and lpa gene from B. subtilis B3 and B. subtilis A13, which produces iturin A, contain an approximate 1-kb nucleotide fragment encoding an aspartate aminotransferase-like protein; however, the relevant regions from B. subtilis 168 and B. subtilis ATCC21332 producing surfactin comprise an approximately 4-kb nucleotide fragment encoding four unknown proteins. There is 73% identity between the Lpa family and the Sfp family, although both are highly conserved.