Aims: To characterize freshwater Bdellovibrio‐and‐like organisms (BALO) isolated in China and examine their potential in controlling growth of Salmonella enterica ssp. enterica serovar Typhimurium on tilapia fillets. Methods and Results: Four BALO isolates were recovered from a pond in Yanzhou of Shandong province, China, with Salm. Typhimurium as prey using double‐layer agar method. Partial 16S rDNA sequencing analysis identified BD2GL, BD5GL and BDXGL as Bdellovibrio bacteriovorus and BD2GS as a Peredibacter sp. Lysis experiments on 32 potentially pathogenic strains revealed that BALO lysis rates are in the range of 56·3–65·6%. On the five Salmonella strains tested, only BD2GS achieved 100% lysis rate. When applied on tilapia fillets against Salm. Typhimurium, BD2GS showed its growth control potential. Cell increments of Salm. Typhimurium were significantly lower (P < 0·05) in two BD2GS‐treated groups compared to control and low‐dose group (BD2GS to prey ratio, 1 : 1) was more effective than high‐dose group (BD2GS to prey ratio, 10 : 1) in controlling Salm. Typhimurium growth. Conclusions: Results of this study indicated that BD2GS could control Salm. Typhimurium growth on tilapia fillets. Significance and Impact of the Study: BALO could be used as a live protective culture in controlling bacterial growth and ensure food safety.
Monacolin J (MJ), a key precursor of Lovastatin, could synthesize important statin drug simvastatin by hydrolyzing lovastatin and adding different side chains. In this study, to reduce the cumbersome hydrolysis of lovastatin to produce MJ in the native strain Aspergillus terreus, the MJ biosynthetic pathway genes (lovB, lovC, lovG, and lovA) were heterologously integrated into the genome of Aspergillus. niger CBS513.88 with strong promoters and suitable integration sites, via yeast 2μ homologous recombination to construct expression cassettes of long-length genes and CRISPR/Cas9 homology-directed recombination (CRISPR-HDR) to integrate MJ genes in the genome of A. niger. RT-PCR results proved that pathway synthesis-related genes could be heterologously expressed in A. niger. Finally, we constructed an engineered strain that could produce monacolin J, detected by LC-HR-ESIMS (MJ, 339.22 [M-H]+). The yield of MJ reached 92.90 mg/L after 7-day cultivation. By optimizing the cultivation conditions and adding precursor, the final titer of MJ was 142.61 mg/L on the fourth day of fed-batch cultivation, which was increased by 53.5% compared to the original growth conditions. Due to the wide application of A. niger in industrial fermentation for food and medicine, the following work will be dedicated to optimizing the metabolic network to improve the MJ production in the engineered strain.
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