A high-throughput screening method based on the Mizoroki-Heck reaction for isolating itaconic acid-producing fungi from soils

Sano, Mei; Kuroda, Hikari; Ohara, Hirotaka; Andō, Hiroshi; Matsumoto, Keiji; Aso, Yoichi

doi:10.1016/j.heliyon.2019.e02048

Cited by 9 publications

(9 citation statements)

References 23 publications

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“…Collectively, the findings demonstrate that agar medium containing 50 mM TG and 0.5 mM VA is the most suitable medium to screen microbes producing vinyl compounds based on the thiol-ene reaction. In combination with another screening method based on the Mizoroki-Heck reaction, which we have reported previously 17 , a novel microbial screening method, DISCOVER, was developed.…”

Section: Resultsmentioning

confidence: 99%

“…The cultures were incubated at 30 °C for 7 days using an incubator M·BR-034P (TAITEC, Saitama, Japan) with shaking at 1600 rpm. The Mizoroki-Heck reaction followed by iodine test (2 nd step of screening) was performed according to the method described elsewhere 17 . Mass determination of vinyl compounds labelled with IB was performed in the same manner as analysis of the reactants of TG with IA.…”

Section: Methodsmentioning

confidence: 99%

“…The Mirozoki-Heck reaction is a coupling reaction between a haloalkane and an alkene that occurs in the presence of palladium catalysts under alkaline conditions. Recently, we developed a screening method for isolating IA producers from soil based on the Mirozoki-Heck reaction and followed by an iodine-test 17 . Using this method, we have isolated A. terreus which produces IA.…”

Section: Introductionmentioning

confidence: 99%

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DISCOVER: A facile structure-based screening method for vinyl compound producing microbes

Aso

Sano

Kuroda

et al. 2019

Sci Rep

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Here we report a novel structure-based microbial screening method for vinyl compound discovery, DISCOVER (direct screening method based on coupling reactions for vinyl compound producers). Through a two-step screening procedure based on selective coupling reactions of terminal alkenes, the thiol-ene reaction (1st step of screening) and Mizoroki-Heck reaction, followed by iodine test (2nd step of screening), microbes producing vinyl compounds like itaconic acid (IA) can be isolated from soil samples. In the 1st step of screening, soil sources are plated on agar medium supplemented with an antimicrobial agent, α-thioglycerol (TG), and a radical initiator, VA-044 (VA). In the 2nd step of screening, vinyl compounds produced in the cultures are labelled with iodobenzene via the Mizoroki-Heck reaction, followed by an iodine test, leading to the detection and characterisation of labelled products. We evaluated the validity of DISCOVER using IA and its producer Aspergillus terreus. Experimental data supported our hypothesis that IA reacts with TG in the medium via the thiol-ene reaction and consequently, A. terreus rapidly forms colonies on the agar medium because of the loss of the antimicrobial activity of TG. Using DISCOVER, high throughput and selective isolation of A. terreus strains producing IA was possible from soils.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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DISCOVER: A facile structure-based screening method for vinyl compound producing microbes

Aso

Sano

Kuroda

et al. 2019

Sci Rep

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“…2019; Sano et al . 2019). As a result, we obtained a new fungus Aspergillus niger S17‐5 (Sano et al .…”

Section: Introductionmentioning

confidence: 99%

“…Recently, we developed a unique microbial screening method based on the Mizoroki-Heck reaction for the isolation of fungi producing IA and its derivatives from soils (Sano et al 2015;Aso et al 2019;Sano et al 2019). As a result, we obtained a new fungus Aspergillus niger S17-5 (Sano et al 2020).…”

Section: Introductionmentioning

confidence: 99%

Caprylic acid enhances hydroxyhexylitaconic acid production in Aspergillus niger S17‐5

et al. 2020

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Aim Aspergillus niger S17‐5 produces two alkylitaconic acids, 9‐hydroxyhexylitaconic acid (9‐HHIA) and 10‐hydroxyhexylitaconic acid (10‐HHIA), which have cytotoxic and polymer building block properties. In this study, we characterized the production of 9‐HHIA and 10‐HHIA by addition of their expected precursor, caprylic acid, to a culture of A. niger S17‐5, and demonstrated batch fermentation of 9‐HHIA and 10‐HHIA in a jar fermenter with DO‐stat. Methods and Results Production titres of 9‐HHIA and 10‐HHIA from 3% glucose in a flask after 25 days cultivation were 0·35 and 1·01 g l−1 respectively. Addition of 0·22 g l−1 of caprylic acid to a suspension of resting cells of A. niger S17‐5 led to 32% enhancement of total 9‐HHIA and 10‐HHIA production compared to no addition. No enhancement of the production of 9‐HHIA or 10‐HHIA by the addition of oxaloacetic acid was observed. Addition of caprylic acid to the culture at mid‐growth phase was more suitable for 9‐HHIA and 10‐HHIA production due to less cell growth inhibition by caprylic acid. DO‐stat batch fermentation with 3% glucose and 14·4 g l−1 of caprylic acid in a 1·5 l jar fermenter resulted in the production titres of 9‐HHIA and 10‐HHIA being 0·48 and 1·54 g l−1 respectively after 10 days of cultivation. Conclusions Addition of caprylic acid to the culture of A. niger S17‐5 enhances 9‐HHIA and 10‐HHIA production. Significance and Impact of the Study These results suggest that 9‐HHIA and 10‐HHIA are synthesized with octanoyl‐CoA derived from caprylic acid, and that the supply of octanoyl‐CoA is a rate‐limiting step in 9‐HHIA and 10‐HHIA production. To the best of our knowledge, this is the first report regarding the fermentation of naturally occurring itaconic acid derivatives in a jar fermenter.

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Photoclick reaction for rapid and simple fluorescence detection of itaconic acid and its derivatives in fungal cultures

Aso,

Tanaka,

Miyazaki

et al. 2023

Anal Bioanal Chem

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A high-throughput screening method based on the Mizoroki-Heck reaction for isolating itaconic acid-producing fungi from soils

Cited by 9 publications

References 23 publications

DISCOVER: A facile structure-based screening method for vinyl compound producing microbes

DISCOVER: A facile structure-based screening method for vinyl compound producing microbes

Caprylic acid enhances hydroxyhexylitaconic acid production in Aspergillus niger S17‐5

Photoclick reaction for rapid and simple fluorescence detection of itaconic acid and its derivatives in fungal cultures

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