Significantly enhanced substrate tolerance of Pseudomonas putida nitrilase via atmospheric and room temperature plasma and cell immobilization

Dong, Ting-Ting; Gong, Jin‐Song; Gu, Bing-Chen; Zhang, Qiang; Li, Heng; Lü, Zhen-Ming; Lu, Maolin; Shi, Jin‐Song; Xu, Zhenghong

doi:10.1016/j.biortech.2017.08.039

Cited by 43 publications

(16 citation statements)

References 34 publications

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“…Over 250 Gy, the fatality rate exceeded 95.47%; at 300 Gy, the active M. purpureus LQ-6 spores were almost completely killed. According to modern theory of breeding and

mutation rate, a 90% fatality rate was set as the target ( Dong et al, 2017 ). Thus, an irradiation dose of 200 Gy was chosen for further evaluation.…”

Section: Resultsmentioning

confidence: 99%

Increased Water-Soluble Yellow Monascus Pigment Productivity via Dual Mutagenesis and Submerged Repeated-Batch Fermentation of Monascus purpureus

et al. 2022

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Monascus pigments (MPs) have been used in the food industry for more than 2,000 years and are known for their safety, bold coloring, and physiological activity. MPs are mainly yellow (YMPs), orange (OMPs), and red (RMPs). In this study, a mutant strain Monascus purpureus H14 with high production of water-soluble YMPs (WSYMPs, λmax at 370 nm) was generated instead of primary YMPs (λmax at 420 nm), OMPs (λmax at 470 nm), and RMPs (λmax at 510 nm) produced by the parent strain M. purpureus LQ-6 through dual mutagenesis of atmospheric and room-temperature plasma and heavy ion beam irradiation (HIBI), producing 22.68 U/ml extracellular YMPs and 10.67 U/ml intracellular YMPs. WSYMP production was increased by 289.51% in optimal conditions after response surface methodology was applied in submerged fermentation. Application of combined immobilized fermentation and extractive fermentation improved productivity to 16.89 U/ml/day, 6.70 times greater than with conservative submerged fermentation. The produced WSYMPs exhibited good tone stability to environmental factors, but their pigment values were unstable to pH, light, and high concentrations of Ca2+, Zn2+, Fe2+, Cu2+, and Mg2+. Furtherly, the produced exYMPs were identified as two yellow monascus pigment components (monascusone B and C21H27NO7S) by UHPLC-ESI-MS. This strategy may be extended to industrial production of premium WSYMPs using Monascus.

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“…Over 250 Gy, the fatality rate exceeded 95.47%; at 300 Gy, the active M. purpureus LQ-6 spores were almost completely killed. According to modern theory of breeding and

mutation rate, a 90% fatality rate was set as the target ( Dong et al, 2017 ). Thus, an irradiation dose of 200 Gy was chosen for further evaluation.…”

Section: Resultsmentioning

confidence: 99%

Increased Water-Soluble Yellow Monascus Pigment Productivity via Dual Mutagenesis and Submerged Repeated-Batch Fermentation of Monascus purpureus

et al. 2022

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“…The individual colonies grown on each mutated medium and the control medium were counted (Dong et al . 2017). The lethality rate was determined as follows:

Lethality rate false(% false) = (1 ‐ \frac{Survival colonies}{Control colonies}) \times 100

…”

Section: Methodsmentioning

confidence: 99%

High‐yield strain of fusidic acid obtained by atmospheric and room temperature plasma mutagenesis and the transcriptional changes involved in improving its production in fungus Fusidium coccineum

Huang

Huang³

et al. 2020

J Appl Microbiol

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Aims To obtain the high‐yield strain of fusidic acid, which is produced from fungus Fusidium coccineum and is the only fusidane‐type antibiotic that has been used clinically, and confirm the changes in the transcription levels involved in increasing its production. Methods and Results By using the atmospheric and room temperature plasma mutagenesis technology, a high‐yield mutant strain of fusidic acid‐producing fungus F. coccineum was obtained. Using the genomic analysis of the original strain based on biosynthetic pathways of ergosterol and helvolic acid, we demonstrate that the pathway involved in the biosynthesis of 2,3‐oxidosqualene from acetyl coenzyme A was shared by fusidic acid and ergosterol, and fusidic acid was finally synthesized by the catalysis of multiple cytochrome P450s and short‐chain dehydrogenase/reductase from 2,3‐oxidosqualene. Then, through the transcriptomic analysis of the original and mutagenized strain, it revealed that the proposed pathway from sucrose to fusidic acid was the most significantly up‐regulated in the transcription levels of the mutant strain. Conclusions The changes in the transcription levels of fusidic acid during its biosynthesis might result in high‐yield of fusidic acid in the mutant strain. This is the first report on the whole biosynthetic pathway of fusidic acid in F. coccineum. Significance and Impact of the Study This study obtain the genetic basis for the biosynthesis of fusidic acid which could be beneficial for the molecular modifications of F. coccineum to further increase its yield by fermentation in future, and established the foundation to reveal the mechanism of the high‐yield of the mutant strain.

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“…Consequently, it has become possible to conduct in-silico mining of new efficient nitrilases from a large number of database resources, which gradually replaced the traditional nitrilase screening strategies. The discovery and screening, 2,[5][6][7] cloning and expression, 8,9 and molecular modification [10][11][12][13] of numerous nitrilases has been reported. In recent years, the expression of nitrilases in recombinant Escherichia coli has increasingly relied on constitutive promoters to avoid the need for adding expensive chemical inducers.…”

Section: Introductionmentioning

confidence: 99%

Constitutive secretory expression and characterization of nitrilase from Alcaligenes faecalis in Pichia pastoris for production of R‐mandelic acid

Zhang

Wang

et al. 2021

Biotech and App Biochem

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Nitrilases can directly hydrolyze nitrile compounds into carboxylic acids and ammonium. To solve the current problems of bioconversions using nitrilases, including the difficult separation of products from the resting cells used as the catalyst and high costs of chemical inducers, a nitrilase from Alcaligenes faecalis was heterologously expressed in Pichia pastoris X33. The stable nitrilaseexpressing strain No.39-6-4 was obtained after three rounds of screening based on a combined detection method including dot-blot, SDS-PAGE, and western blot analyses, which confirmed the presence of recombinant nitrilase with a molecular mass of about 50 kDa. The temperature and pH optima of the nitrilase were 45 • C and pH 7.5, respectively. Cu 2+ , Zn 2+ , and Tween 80 strongly inhibited the enzyme activity, but the optical purity of the product R-mandelic acid (R-MA) was stable, with practically 100% enantiomeric excess (ee). The nitrilaseproducing P. pastoris strain developed in this study provides a basis for further research on the enzyme.

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Significantly enhanced substrate tolerance of Pseudomonas putida nitrilase via atmospheric and room temperature plasma and cell immobilization

Cited by 43 publications

References 34 publications

Increased Water-Soluble Yellow Monascus Pigment Productivity via Dual Mutagenesis and Submerged Repeated-Batch Fermentation of Monascus purpureus

Increased Water-Soluble Yellow Monascus Pigment Productivity via Dual Mutagenesis and Submerged Repeated-Batch Fermentation of Monascus purpureus

High‐yield strain of fusidic acid obtained by atmospheric and room temperature plasma mutagenesis and the transcriptional changes involved in improving its production in fungus Fusidium coccineum

Constitutive secretory expression and characterization of nitrilase from Alcaligenes faecalis in Pichia pastoris for production of R‐mandelic acid

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