Functional characterization of a novel violacein biosynthesis operon from Janthinobacterium sp. B9-8

Xu, Xing‐You; Chu, Xiaoting; Du, Bangmian; Huang, Chun-Yan; Xie, Chengzhi; Zhang, Zhidong; Jiang, Ling

doi:10.1007/s00253-022-11929-8

Cited by 7 publications

(7 citation statements)

References 57 publications

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“…39 The single colony of VioABCDE-SD grown on an LB plate (10 g/L tryptone, 5 g/L yeast extract, 10 g/L sodium chloride, 20 g/L agar) was inoculated in 50 mL of LB broth with addition of ampicillin (100 mg/mL) and kanamycin (10 mg/mL) at 37 • C, 200 rpm for 16 h. Then the seed cultures were then transferred to fresh TGY medium (5 g/L tryptone, 3 g/L yeast extract, 1 g/L glucose) at an inoculation amount of 4% and cultured at 37 • C until the OD 600 reached 0.8-1.0. Then lactose (0.8 mM) was supplemented and the culture condition was changed to 15 • C and pH 4.0 for 108 h with the addition of tryptophan (1.2 mM) at 8 h. 39 Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Deinococcus radiodurans R1, and E. coli BL21 were preserved in our laboratory. For antimicrobial tests, the single colony of S. aureus, B. subtilis, P. aeruginosa, or E. coli was incubated at 37 • C, 200 rpm for 12 h in LB liquid medium.…”

Section: Strains and Culture Methodsmentioning

confidence: 99%

“…Engineered E. coli VioABCDE‐SD was constructed in the previous study 39 . The single colony of VioABCDE‐SD grown on an LB plate (10 g/L tryptone, 5 g/L yeast extract, 10 g/L sodium chloride, 20 g/L agar) was inoculated in 50 mL of LB broth with addition of ampicillin (100 mg/mL) and kanamycin (10 mg/mL) at 37°C, 200 rpm for 16 h. Then the seed cultures were then transferred to fresh TGY medium (5 g/L tryptone, 3 g/L yeast extract, 1 g/L glucose) at an inoculation amount of 4% and cultured at 37°C until the OD 600 reached 0.8–1.0.…”

Section: Methodsmentioning

confidence: 99%

“…The mobile phase consisted of acetonitrile–water (4:6, V/V) and the flow rate was 1.0 mL/min. The injection volume was set to 20 μL 39 . Ultra‐performance liquid chromatography (UPLC) analysis was performed with an ACQUITY UPLC BEH‐C18 column (2.1 × 100 mm, 1.7 μm; Waters, USA).…”

Section: Methodsmentioning

confidence: 99%

“…The injection volume was set to 20 μL. 39 Ultra-performance liquid chromatography (UPLC) analysis was performed with an ACQUITY UPLC BEH-C18 column (2.1 × 100 mm, 1.7 μm; Waters, USA). The mass spectra were recorded using A SYNAPT G2-Si mass spectrometer (Waters).…”

Section: Extraction and Analysis Of Violaceinmentioning

confidence: 99%

“…B9-8 was identified and expressed in E.coli to obtain an engineered strain VioABCDE-SD with a high violacein titer in our previous study. 39 In this paper, a rapid and efficient method for extracting violacein produced by engineered E. coli VioABCDE-SD was established and the stability, antimicrobial activity, and antioxidant capacity of produced violacein were evaluated and compared. We hope to find the uniqueness of violacein synthesized from Janthinobacterium sp.…”

Section: Introductionmentioning

confidence: 99%

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Study on extraction and characterization of new antibiotics violacein from engineered Escherichia coli VioABCDE‐SD

Huang

Chu

Hui

et al. 2023

Biotech and App Biochem

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To better understand the characteristic properties of violacein biosynthesized by engineered Escherichia coli VioABCDE‐SD, a convenient and simplified method was designed to extract violacein and its stability, antimicrobial activity, and antioxidant capacity were analyzed. Different from the traditional extraction methods, our new method is easier and less time consuming and can directly obtain violacein dry powder product with a higher extraction rate. Low temperature, dark condition, neutral pH, reducing agents, Ba2+, Mn2+, Ni2+, Co2+, and some food additives of sucrose, xylose, and glucose were conducive to maintaining its stability. The violacein also exhibited surprisingly high bacteriostatic action against Gram‐positive Bacillus subtilis, Deinococcus radiodurans R1, and Staphylococcus aureus and Gram‐negative Pseudomonas aeruginosa, but no effect on E. coli. The violacein of VioABCDE‐SD exhibited strong antioxidant activity, with the scavenging rate of 1,1‐diphenyl‐2‐picrylhydrazyl free radicals reaching 60.33%, the scavenging efficiency of hydroxyl radical scavenging reaching 56.34%, and the total antioxidant capacity reaching 0.63 U/mL. Violacein from VioABCDE‐SD can be synthesized directionally with better stability, antibacterial, and antioxidant properties compared with that from the original strain Janthinobacterium sp. B9‐8. Therefore, our study indicated that violacein from engineered E. coli VioABCDE‐SD was a kind of new antibiotic with potential biological activities, which may have potential utility in multiple areas such as pharmacological, cosmetics, and healthy food industries.

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Section: Strains and Culture Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Extraction and Analysis Of Violaceinmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Study on extraction and characterization of new antibiotics violacein from engineered Escherichia coli VioABCDE‐SD

Huang

Chu

Hui

et al. 2023

Biotech and App Biochem

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Stability of a Mutualistic Escherichia coli Co‐Culture During Violacein Production Depends on the Kind of Carbon Source

Schick,

Müller,

Takors

et al. 2024

Engineering in Life Sciences

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The L‐tryptophan–derived purple pigment violacein (VIO) is produced in recombinant bacteria and studied for its versatile applications. Microbial synthetic co‐cultures are gaining more importance as efficient factories for synthesizing high‐value compounds. In this work, a mutualistic and cross‐feeding Escherichia coli co‐culture is metabolically engineered to produce VIO. The strains are genetically modified by auxotrophies in the tryptophan (TRP) pathway to enable a metabolic division of labor. Therein, one strain produces anthranilate (ANT) and the other transforms it into TRP and further to VIO. Population dynamics and stability depend on the choice of carbon source, impacting the presence and thus exchange of metabolites as well as overall VIO productivity. Four carbon sources (D‐glucose, glycerol, D‐galactose, and D‐xylose) were compared. D‐Xylose led to co‐cultures which showed stable growth and VIO production, ANT‐TRP exchange, and enhanced VIO production. Best titers were ∼126 mg L–1 in shake flasks. The study demonstrates the importance and advantages of a mutualistic approach in VIO synthesis and highlights the carbon source's role in co‐culture stability and productivity. Transferring this knowledge into an up‐scaled bioreactor system has great potential in improving the overall VIO production.

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Pigments from pathogenic bacteria: a comprehensive update on recent advances

Acharya,

Shaw,

Bhattacharya

et al. 2024

World J Microbiol Biotechnol

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Functional characterization of a novel violacein biosynthesis operon from Janthinobacterium sp. B9-8

Cited by 7 publications

References 57 publications

Study on extraction and characterization of new antibiotics violacein from engineered Escherichia coli VioABCDE‐SD

Study on extraction and characterization of new antibiotics violacein from engineered Escherichia coli VioABCDE‐SD

Stability of a Mutualistic Escherichia coli Co‐Culture During Violacein Production Depends on the Kind of Carbon Source

Pigments from pathogenic bacteria: a comprehensive update on recent advances

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