Improved the lipopeptide production of Bacillus amyloliquefaciens HM618 under co-culture with the recombinant Corynebacterium glutamicum producing high-level proline

Chen, Xinyue; Sun, Hongmin; Qiao, Bin; Miao, Chang-Hao; Hou, Zheng-Jie; Xu, Shujing; Xu, Qiu-Man; Cheng, Jing-Sheng

doi:10.1016/j.biortech.2022.126863

Cited by 27 publications

(22 citation statements)

References 44 publications

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“…Several studies have evaluated the effects of media (e.g., carbon, nitrogen sources, and metal ions) on the production of secondary metabolites. ,, As shown in Figure A1, B1, we determined the effects of media Cal18 and YBD on fengycin production. The Cal18 medium significantly increased the growth and fengycin production of strains BSf01 and BSf26 compared with the YBD medium.…”

Section: Resultsmentioning

confidence: 99%

“…The antifungal activity of fengycin was assayed by the Oxford cup method with Botrytis cinerea and Rhizoctonia solani as the indicator fungi. 45 About 1 cm of mycelia agar from B. cinerea or R. solani was placed on the center of a plate containing potato dextrose agar (PDA) media. After incubation at 30 °C for 24−36 h, 150.0 μL fengycin extract was added to the Oxford cup.…”

Section: Extraction and Identification Of Fengycinmentioning

confidence: 99%

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Improved Production of Fengycin in Bacillus subtilis by Integrated Strain Engineering Strategy

et al. 2022

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Fengycin is a lipopeptide with broad-spectrum antifungal activity. However, its low yield limits its commercial application. Therefore, we iteratively edited multiple target genes associated with fengycin synthesis by combinatorial metabolic engineering. The ability of Bacillus subtilis 168 to manufacture lipopeptides was restored, and the fengycin titer was 1.81 mg/L. Fengycin production was further increased to 174.63 mg/L after knocking out pathways associated with surfactin and bacillaene synthesis and replacing the native promoter (P ppsA ) with the P veg promoter. Subsequently, fengycin levels were elevated to 258.52 mg/L by upregulating the expression of relevant genes involved in the fatty acid pathway. After blocking spore and biofilm formation, fengycin production reached 302.51 mg/L. Finally, fengycin production was increased to approximately 885.37 mg/L after adding threonine in the optimized culture medium, which was 488-fold higher compared with that of the initial strain. Integrated strain engineering provides a strategy to construct a system for improving fengycin production.

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Section: Resultsmentioning

confidence: 99%

Section: Extraction and Identification Of Fengycinmentioning

confidence: 99%

Improved Production of Fengycin in Bacillus subtilis by Integrated Strain Engineering Strategy

et al. 2022

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“…In order to improve the production of plipastatin, several strategies have been developed in recent years. Chen et al increased the yield of plipastatin by 28% by adding proline to the culture medium and achieved 11.41 mg/L . Zhou et al achieved a 79% increase in plipastatin yield, resulting in a yield of 327.598 mg/L, through the knockout of the negative regulatory factor FenSr3 of plipastatins .…”

Section: Introductionmentioning

confidence: 99%

“…Chen et al increased the yield of plipastatin by 28% by adding proline to the culture medium and achieved 11.41 mg/L. 11 Zhou et al achieved a 79% increase in plipastatin yield, resulting in a yield of 327.598 mg/ L, through the knockout of the negative regulatory factor FenSr3 of plipastatins. 12 The deletion of a 6 kb gene sequence of surfactin BC by Zou et al led to a plipastatin yield of 1600 mg/L.…”

Section: Introductionmentioning

confidence: 99%

Research on the Regulation of Plipastatin Production by the Quorum-Sensing ComQXPA System of Bacillus amyloliquefaciens

Zhou

Zheng

et al. 2023

J. Agric. Food Chem.

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Plipastatin is a cyclic lipopeptide synthesized by non-ribosomal peptide synthetases (NRPS), which has a diverse range of applications in postharvest preservation of fruits and vegetables, biological control, and feed processing. Whereas the yield of plipastatin in wild Bacillus sp. is low, its chemical structure is complex and challenging to synthesize, significantly limiting its production and application. ComQXPA-PsrfA, a quorum-sensing (QS) circuit from Bacillus amyloliquefaciens, was constructed in this study. Two QS promoters MuPsrfA and MtPsrfA, with 35 and 100% increased activity, respectively, were obtained by mutating the original promoter PsrfA. Thus, the natural promoter of plipastatin was replaced by a QS promoter to achieve the dynamic regulation of plipastatin, which increased the yield of plipastatin by 3.5 times. Integrating ComQXPA into plipastatin mono-producing M-24:MtPsrfA increased the yield of plipastatin to 3850 mg/L, representing the highest yield reported to date. Four new plipastatins were identified via UPLC-ESI-MS/MS and GC–MS analysis of fermentation products of mono-producing engineered strains. Among them, three plipastatins contained two double bonds in the fatty acid side chain, representing the first example of a new type of plipastatin. Our results indicate that the QS system ComQXPA-PsrfA of Bacillus can dynamically regulate plipastatin production, and the pipeline could be extended to the other strains to regulate target products dynamically.

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“…Streptomyces and their metabolites have long been an important resource for the biological control of plant diseases, and their beneficial ecological effects are related to their ability to produce a wide range of antimicrobial compounds, including non-peptide compounds such as polyketides, amino glycans, phospholipids, and peptide compounds such as cyclic lipopeptides (CLPs). CLPs are cyclic peptides with amino acid residues and fatty acid tails of varying lengths, which are synthesized through nonribosomal peptide synthetase [ 5 ]. This large antimicrobial group is composed of three main families: fengycins, iturins and surfactins [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Transcriptome analysis of the production enhancement mechanism of antimicrobial lipopeptides of Streptomyces bikiniensis HD-087 by co-culture with Magnaporthe oryzae Guy11

et al. 2022

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The lipopeptides produced by Streptomyces bikiniensis have a significant inhibitory effect on Magnaporthe oryzae, but the low yield limits its application. In this study, the anti-M. oryzae activity of the broth of S. bikiniensis HD-087 co-cultured with M. oryzae Guy11 mycelium has risen by 41.22% compared with pure culture, and under induction conditions of adding Guy11-inducer (cell-free supernatant of M. oryzae Guy11), the activity of strain HD-087 improved 61.76%. The result proved that the enhancement effect of Guy11 on the antimicrobial activity of HD-087 was mainly related to metabolites but mycelium cells. Under optimum induction conditions, NRPS gene expression levels of HD-087 were significantly increased by induction with Guy11-inducer, the biomass of HD-087 had no significant change, but crude extract of lipopeptide (CEL) production was 107.4% higher than pure culture, and TLC result under acid hydrolysis showed that the induced culture has one component more than pure culture. To clarify the regulation mechanism of improving lipopeptide production of HD-087 with Guy11-inducer, transcriptomic analysis was performed using RNAseq to compare the induced culture and pure culture. In the induced culture, 943 genes were up-regulated, while 590 genes were down-regulated in DEGs (differentially expressed genes). KEGG results showed that the expression of genes related to amino acid synthesis, fatty acid metabolism, TCA cycle and pyruvate metabolism pathway were significantly increased. The increased expression of genes related to these metabolic pathways provided sufficient precursors for lipopeptide synthesis. Accordingly, key enzyme genes responsible for the synthesis of lipopeptides Srf and NRPS was significantly increased. Quorum sensing related genes OppA and MppA were significantly up-regulated, and then ComP was activated and promoted lipopeptide synthesis. These results provided a scientific basis for using M. oryzae to induce the increase of the production of Streptomyces lipopeptides, and also laid a foundation for further exploring the co-culture mechanisms among different genera.

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Improved the lipopeptide production of Bacillus amyloliquefaciens HM618 under co-culture with the recombinant Corynebacterium glutamicum producing high-level proline

Cited by 27 publications

References 44 publications

Improved Production of Fengycin in Bacillus subtilis by Integrated Strain Engineering Strategy

Improved Production of Fengycin in Bacillus subtilis by Integrated Strain Engineering Strategy

Research on the Regulation of Plipastatin Production by the Quorum-Sensing ComQXPA System of Bacillus amyloliquefaciens

Transcriptome analysis of the production enhancement mechanism of antimicrobial lipopeptides of Streptomyces bikiniensis HD-087 by co-culture with Magnaporthe oryzae Guy11

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