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Salmonella pullorum phage Pu29 was comprehensively analyzed for its biological and genomic characteristics. A magnetic separation and enrichment technique for Salmonella was established using the phage Pu29 as a recognition element. The phage belonged to the genus Roufvirus and had an icosahedron head and an irreducible long tail. Pu29 had a wide host spectrum with an adsorption rate of 88.67% on host cells in 15 min, a latent period of 30 min, a rise period of 180 min, and a burst size of 115.74 PFU/cell. Meanwhile, Pu29 had good heat resistance (30–60 ℃) and pH tolerance (pH 4–11). Its genome was composed of 45 715 bp (GC content 46.08%) and 81 open reading frames (ORFs), including 18 ORFs with known functions that did not carry genes encoding toxicity or resistance factors. PhagePu29-MBs were prepared as a probe by coupling the phage with carboxylated nano-magnetic beads (MBs) through amide reaction. When 25 μg of the probe was incubated with Salmonella at 37 ℃ for 20 min, the highest capture rate of Salmonella of 83.93% and the lowest captured bacterial concentration of 45 CFU/mL were obtained. Transmission electron microscopy (TEM) was used to observe that PhagePu29-MBs could specifically capture Salmonella. In spiked samples, the highest capture rate of Salmonella separated and enriched by the probe reached 92.92%. The separation and enrichment process took approximately 30 min. Therefore, this study established a fast and highly specific magnetic separation method for Salmonella based on phage Pu29, which may lay the foundation for the development of a phage-based method for the rapid separation and enrichment of foodborne pathogens in food samples.

Esa1 has been proven to be an important histone acetyltransferase involved in the regulation of growth and metabolism. Monascus spp. with nearly 2000 years of edible history in East Asian countries can produce a variety of polyketides. It is unknown whether Esa1 plays a regulatory role in Monascus spp. In this study, we isolated the homology of histone acetyltransferase Esa1 (named MrEsa1) and constructed a mresa1‐overexpressed strain. Western blot experiments showed that MrEsa1 hyperacetylated at K4 and K9 of the H3 subunit in Monascus ruber. Overexpression of mresa1 led to the larger colony diameter and increased dry cell mass; meanwhile, the conidia germination rate was significantly accelerated in the mresa1‐overexpressed strain before 4 h, and the number of ascospores in the mresa1‐overexpressed strain was significantly higher than that in WT. In addition, the Monascus azaphilone pigments (MonAzPs) and citrinin production of the mresa1‐overexpressed strain were 1.7 and 2.4 times more than those of WT, respectively. Reverse transcription‐quantitative polymerase chain reaction experiment suggested that mrpigB, mrpigH, mrpigJ, and mrpigK, involved in MonAzPs synthesis, and pksCT, mrl3, and mrl7, involved in citrinin synthesis, were upregulated in mresa1‐overexpressed strain. This study provides important insights into the effect of MrEsa1 on the developmental process and the production of secondary metabolites in Monascus spp. [ABSTRACT FROM AUTHOR]

Aims In this study, Mrhst4 , encoding a member of NAD+-dependent histone deacetylase (HDAC), was deleted to evaluate its regulation on the production of Monascus azaphilone pigments (MonAzPs) and mycotoxin, as well as the developmental process in Monascusruber. Methods and results Agrobacterium tumefaciens -mediated transformation was applied in this study to generate the Mrhst4 null strain. Mrhst4 -deleted strain did not display obvious differences in the sexual and asexual reproduction, colonial morphology, and micro-morphology. UV–Vis scan and UPLC detection showed that disruption of Mrhst4 significantly increased the MonAzPs yields, and citrinin content was dramatically enhanced during the tested period. RT-qPCR results showed that the absence of Mrhst4 significantly increased the relative expression of citrinin biosynthetic pathway genes including pksCT, mrl1, mrl2, mrl4, mrl6 , and mrl7. The Western blot assay suggested that deletion of Mrhst4 could significantly elevate the acetylation levels of H3K4, H3K9, H3K18, H3K56, and H4K12, but attenuated the lysine acetylation modification of H4Pan, H4K8, and H4K16. Conclusion MrHst4 is an important regulator involved in secondary metabolism in Monascus ruber. In particular, MrHst4 plays a pivotal role in regulation of citrinin production. [ABSTRACT FROM AUTHOR]