Antisense RNA-Based Strategy for Enhancing Surfactin Production in <i>Bacillus subtilis</i> TS1726 via Overexpression of the Unconventional Biotin Carboxylase II To Enhance ACCase Activity

Wang, Miaomiao; Yu, Huimin; Shen, Zhongyao

doi:10.1021/acssynbio.8b00459

Cited by 27 publications

(13 citation statements)

References 30 publications

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“…Overexpression of accDA likely intensifies the inverse process response, thus causing a decrease in fengycin production. The ldeHA gene encodes acetyl-CoA carboxylase and is associated with the formation of malonyl-CoA from acetyl-CoA . Compared with the control (strain BSf04-2), the biomass of strain BSf07 overexpressing ldeHA with the P43 promoter was elevated (Figure A).…”

Section: Resultsmentioning

confidence: 99%

“…The ldeHA gene encodes acetyl-CoA carboxylase and is associated with the formation of malonyl-CoA from acetyl-CoA. 36 Compared with the control (strain BSf04-2), the biomass of strain BSf07 overexpressing ldeHA with the P43 promoter was elevated (Figure 4A). Meanwhile, the byproduct acetoin was also significantly increased (Figure S2); however, fengycin production of strain BSf07 was significantly decreased (Figure 4B).…”

Section: Effects Of Different Promoters On Fengycin Synthesis In B Su...mentioning

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: Effects Of Different Promoters On Fengycin Synthesis In B Su...mentioning

confidence: 99%

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

et al. 2022

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“…In addition, as reported, biotin carboxylase II, encoded by ldeHA, played a significant role in maintaining ACC activity, and its overexpression was shown to effectively increase the activity of ACC and promote fatty acid biosynthesis. 37 Consequently, we used a strategy to overexpress the genes ldeHA and acc by fusing with the P43 promoter and constructed engineered strains TDS1 and TDS2, respectively. As shown in Figure 2A, the obtained strains TDS1 and TDS2 produced surfactins with yields of 1.28 and 1.25 g/L, respectively, which were increased by 44% and 40% compared to B. subtilis TD7.…”

Section: Effect Of Single-gene Regulation On Surfactin Productionmentioning

confidence: 99%

Genetic engineering of the branched‐chain fatty acid biosynthesis pathway to enhance surfactin production from Bacillus subtilis

Jing

Wei

Liu

et al. 2022

Biotech and App Biochem

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Surfactin, which is composed of a β‐hydroxy fatty acid chain and a peptide ring, has drawn considerable attention due to its potential applications in the biomedicine, bioremediation, and petroleum industries. However, the low yield of surfactin from wild strains still restricts its industrial applications. In this study, eight genes relevant to the fatty acid biosynthesis pathway were targeted to enhance surfactin production, and high surfactin‐yielding strains with potential industrial applications were obtained. When ldeHA and acc were co‐overexpressed, the surfactin yield of recombinant strains TDS8 and TPS8 increased to 1.55‐ and 1.19‐fold of their parental strains, respectively, again proving that the conversion of acetyl‐coenzyme A (CoA) to malonyl‐CoA is the rate‐limiting step in fatty acid biosynthesis. Furthermore, changes in surfactin isoforms of recombinant strain TPS8 suggest that the fatty acid precursor synthesis pathway can be modified to improve the proportion of different isoforms. In addition, the deletion of lpdV, which is responsible for the conversion of α‐ketoacyl‐CoA precursors, resulted in a sharp decrease in surfactin production, further demonstrating the importance of branched‐chain fatty acid biosynthesis in surfactin production. This work will facilitate the design and construction of more efficiently engineered strains for surfactin production and further extend industrial applications.

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“…Branched chain amino acid (BCAA) served as the critical role in erythromycin production, and strengthening BCAA supplies led to a 41% increase of erythromycin yield, via deleting lrp and overexpressing BCAA transporter SACE_5387 (Liu et al, 2017). Moreover, acety-CoA carboxylase was overexpressed to improve malonyl-CoA supply, which further led to a 43% increase of surfactin production, reached 13.37 g/L (Wang et al, 2019). In addition, efficient use of substrate is also critical for metabolite production.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Bacitracin Production by Systematically Engineering S-Adenosylmethionine Supply Modules in Bacillus licheniformis

Cai

Zhang

Zhu

et al. 2020

Front. Bioeng. Biotechnol.

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Bacitracin is a broad-spectrum veterinary antibiotic that widely used in the fields of veterinary drug and feed additive. S-Adenosylmethionine (SAM) is a critical factor involved in many biochemical reactions, especially antibiotic production. However, whether SAM affects bacitracin synthesis is still unknown. Here, we want to analyze the relationship between SAM supply and bacitracin synthesis, and then metabolic engineering of SAM synthetic pathway for bacitracin production in Bacillus licheniformis. Firstly, our results implied that SAM exogenous addition benefited bacitracin production, which yield was increased by 12.13% under the condition of 40 mg/L SAM addition. Then, SAM synthetases and Methionine (Met) synthetases from B. licheniformis, Corynebacterium glutamicum, and Saccharomyces cerevisiae were screened and overexpressed to improve SAM accumulation, and the combination of SAM synthetase from S. cerevisiae and Met synthetase from B. licheniformis showed the best performance, and 70.12% increase of intracellular SAM concentration (31.54 mg/L) and 13.08% increase of bacitraicn yield (839.54 U/mL) were achieved in resultant strain DW2-KE. Furthermore, Met transporters MetN and MetP were, respectively, identified as Met exporter and importer, and bacitracin yield was further increased by 5.94% to 889.42 U/mL via deleting metN and overexpressing metP in DW2-KE, attaining strain DW2-KENP. Finally, SAM nucleosidase gene mtnN and SAM decarboxylase gene speD were deleted to block SAM degradation pathways, and bacitracin yield of resultant strain DW2-KENPND reached 957.53 U/mL, increased by 28.97% compared to DW2. Collectively, this study demonstrated that SAM supply served as the critical role in bacitracin synthesis, and a promising strain B. licheniformis DW2-KENPND was attained for industrial production of bacitracin.

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Antisense RNA-Based Strategy for Enhancing Surfactin Production in Bacillus subtilis TS1726 via Overexpression of the Unconventional Biotin Carboxylase II To Enhance ACCase Activity

Cited by 27 publications

References 30 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

Genetic engineering of the branched‐chain fatty acid biosynthesis pathway to enhance surfactin production from Bacillus subtilis

Enhanced Bacitracin Production by Systematically Engineering S-Adenosylmethionine Supply Modules in Bacillus licheniformis

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