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Recently, we developed a unique microbial screening method based on the Mizoroki–Heck reaction for itaconic acid (IA)-producing fungi. This method revealed that 37 out of 240 fungal strains isolated from soils produce vinyl compounds, including IA. In this study, we further characterized these compounds in order to verify that the screening method permits the isolation of fungi that produce other vinyl compounds, excluding IA. HPLC analysis showed that 11 out of 37 isolated strains produced IA, similar to Aspergillus terreus S12-1. Surprisingly, the other 8 isolated strains produced two vinyl compounds with HPLC retention times different from that of IA. From these strains, the vinyl compounds of Aspergillus niger S17-5 were characterized. Mass spectrometric and NMR analyses showed that they were identical to 8-hydroxyhexylitaconic acid (8-HHIA) and 9-HHIA. This finding showed that 8-HHIA- and 9-HHIA-producing fungi, as well as IA-producing fungi, are ubiquitously found in soils. Neither 8-HHIA nor 9-HHIA showed antibacterial or anti-inflammatory activities. Interestingly, 8-HHIA and 9-HHIA showed cytotoxicity against the human cervical cancer cell line (HeLa) and human diploid cell line (MRC-5), and MRC-5 only, respectively, compared to IA at the same concentration. This study indicates that the screening method could easily discover fungi producing 8-HHIA and 9-HHIA in soils.

Here, we report the photosynthetic production of itaconic acid (IA), a promising building block, from carbon dioxide (CO₂) by Synechocystis sp. PCC6803. The engineered PCC6803 strain expressing cis-aconitate decarboxylase, the key enzyme in IA biosynthesis, produced 0.9 mg/L and 14.5 mg/L of IA at production rates of 42.8 μgL(-1)day(-1) and 919.0 μgL(-1)day(-1), under conditions of constant bubbling with air and 5% CO₂, respectively. This is the first report on the possibility of IA production from CO₂ via the photosynthetic process in cyanobacteria.

In this study, we report a novel method based on the Mizoroki-Heck reaction followed by an iodine test for the screening of itaconic acid-producing fungi from soils. This method is simple, rapid, and requires 10 μL of culture; results are obtained within 1.5 h. The detection limit of itaconic acid in the cultures was 0.13 mM. This is the first report on the direct screening of itaconic acid-producing fungi using a coupling reaction.

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