Kenjiro Sugiyama

As the prevalence of type 2 diabetes increases rapidly, the demand for natural origin dipeptidyl peptidase-IV (DPP-IV) inhibitors with fewer side effects is increasing. In this study, protein-rich Limnospira platensis was investigated as a source of bioactive peptides to enhance its value as a functional food ingredient. Hydrolysates were prepared using three food-processing proteases, individually and in two-step combinations, followed by in silico analysis and peptide identification via liquid chromatography–tandem mass spectrometry. Subsequently, the thermal stability, gastrointestinal resistance, and intestinal permeability (using Caco-2 cells) of the identified peptides were evaluated. It was revealed that the Orientase 22BF digest exhibited high DPP-IV inhibitory activity. From the digest, three novel peptides—SPSPN (IC₅₀ = 144.1 ± 2.2 μM), VPSV (IC₅₀ = 93.6 ± 5.8 μM), and IPIGG (IC₅₀ = 13.4 ± 2.3 μM)—were identified, exhibiting DPP-IV inhibitory potencies comparable to or higher than previously reported Limnospira-derived peptides. Although VPSV exhibited low epithelial permeability (P_app = 4.02 ± 0.69 × 10⁻⁸ cm/s), it remained stable under simulated gastrointestinal digestion conditions, suggesting potential local luminal inhibitory activity within the small intestine. Overall, these findings highlight Limnospira-derived VPSV as a promising functional ingredient candidate with high bioactivity and digestive stability.

Raisin syrup sourdough is a popular traditional leavening method in Japan, yet its specific impact on bread aroma evolution and shelf life stability remains scientifically underexplored. This study characterized the fermentation dynamics and volatile profiles of raisin syrup sourdough bread compared to a commercial yeast control over a 3-day shelf life, utilizing comprehensive two-dimensional gas chromatography–mass spectrometry (GC × GC-TOFMS) and primary metabolite profiling of sugars, amino acids, and organic acids. The analysis resolved over 760 volatiles and revealed a fundamental kinetic divergence. While the yeast control exhibited a 24 h metabolic lag, the raisin sourdough achieved rapid activation, establishing a higher initial volatile load immediately post-baking. Driven by lactic acid bacteria dominance and extensive proteolysis, the sourdough’s acidic environment facilitated the retention of fruity esters and malty branched-chain aldehydes while effectively suppressing lipid oxidation markers like 9,17-Octadecadienal. Key aromatic markers, including benzenepropanol and Octanoate , were significantly elevated and stabilized in the sourdough group. These findings demonstrate that raisin syrup fermentation generates a superior, stable aromatic profile, providing a scientific basis for optimizing clean-label artisan bread production in the Japanese market.

Raisin syrup sourdough is a popular traditional leavening method in Japan, yet its specific impact on bread aroma evolution and shelf-life stability remains scientifically underex-plored. This study characterized the fermentation dynamics and volatile profiles of raisin syrup sourdough bread compared to a commercial yeast control over a 3-day shelf life, utilizing comprehensive two-dimensional gas chromatography–mass spectrometry (GC×GC-TOFMS) and primary metabolite profiling of sugars, amino acids, and organic acids. The analysis resolved over 760 volatiles and revealed a fundamental kinetic divergence: while the yeast control exhibited a 24-hour metabolic lag, the raisin sourdough achieved rapid activation, establishing a higher initial volatile load immediately post-baking. Driven by Lactic Acid Bacteria dominance and extensive proteolysis, the sourdough’s acidic environment facilitated the retention of fruity esters and malty branched-chain aldehydes while effectively suppressing lipid oxidation markers like 9,17-Octadecadienal. Key aromatic markers, including Benzenepropanol and Octanoate , were significantly elevated and stabilized in the sourdough group. These findings demonstrate that raisin syrup fermentation generates a superior, stable aromatic profile, providing a scientific basis for optimizing clean-label artisan bread production in the Japanese market.

Background: Myxol, a monocyclic carotenoid with β- and ψ-end groups, has been identified in only a limited number of bacteria, such as flavobacteria and cyanobacteria. Despite its biological significance, the biosynthetic pathway of myxol is not well understood, and studies on its physiological functions and biological activities are limited because of its rarity. Methods: BLAST homology searches for carotenoid biosynthesis genes in the genome of Nonlabens were performed. The carotenogenesis-related genes in the genome of the marine flavobacteria Nonlabens spongiae were individually cloned and functionally characterized using a heterologous Escherichia coli expression system. Carotenoids from N. spongiae were identified using an LC-MS analysis. Results: We identified a gene cluster involved in carotenoid biosynthesis in the genome of N. spongiae. This cluster includes genes encoding phytoene synthase (CrtB), phytoene desaturase (CrtI), lycopene cyclase (CrtY), carotenoid 1,2-hydratase (CruF), carotenoid 3,4-desaturase (ψ-end group) (CrtD), carotenoid 2-hydroxylase (ψ-end group) (CrtA-OH), and carotene hydro-xylase (CrtZ). Based on the characteristics of these enzymes, the primary products were predicted to be myxol and/or zeaxanthin. A spectroscopic analysis confirmed that myxol was the primary carotenoid. Furthermore, a plasmid containing a reconstructed gene cluster and geranylgeranyl pyrophosphate synthase (CrtE) located outside the cluster was introduced into E. coli. This system predominantly accumulated myxol, indicating that the reconstructed gene cluster enabled efficient myxol production in E. coli. Conclusions: This study highlighted the potential biotechnological applications of the carotenoid biosynthesis gene clusters for myxol production.

The digestion behavior of a food bolus comprising bread particles in the presence of gastric peristalsis remains poorly understood. This study systematically investigated the effect of bread type on in vitro gastric digestion behavior using a human gastric digestion simulator (GDS) that is capable of quantitatively simulating gastric peristalsis. A food bolus consisting of 60 g of bread (white bread, bagel, German bread, French bread, or croissant), 15 mL of a simulated salivary fluid, and 240 mL of a simulated gastric fluid was used for gastric digestion in vitro using the GDS for 3 h at 37 °C. Direct observation of the gastric digestion behavior in the GDS vessel demonstrated that the structure and composition of breads considerably influenced the physical digestion processes of bread particles. These processes include their fracture, rubbing, disintegration, swelling owing to the penetration of gastric fluid, and release of fat from their surface. Fluorescence microscopy enabled an improved understanding of the variations in the microstructure and major component distribution of the breads during the gastric digestion in vitro. The results showed how the different breads influenced gastric digestibility in vitro through quantitative gastric peristalsis. The GDS can also be applicable to studying gastric digestibility in vitro of other types of bread.

Gluten-free rice flour noodles with a flavor and texture profile preferred by the Japanese people were developed. The rice noodles contained potato starch (PS) as a binder. “Koshihikari” was selected from several candidate varieties based on its pasting properties. Since the Japanese people prefer the chewy texture of wheat flour “Udon” noodles, first, the stress−strain characteristics of “Udon” noodles in Japan were quantified, using a mechanical test. Next, different formulations of rice noodles were prepared by changing the amount of PS blended into the noodles. The mechanical tests on wheat and rice noodles show that rice noodles made from 85% rice flour and 15% PS have a texture similar to that of “Udon” noodles. Brown rice noodles containing roasted brown rice flour were also developed. Since brown rice flour hinders the binding of the dough, it was necessary to increase the amount of PS to increase the binding of roasted brown rice flour. Finally, noodles with 70% white rice flour, 10% brown rice flour, and 20% PS were produced. The gas chromatography−mass spectrometry analysis of the volatile compounds contained in white rice noodles and brown rice noodles identified the volatile compounds characteristic each of type.

For carotenogenesis, two biosynthetic pathways from phytoene to lycopene are known. Most bacteria and fungi require only phytoene desaturase (PDS, CrtI), whereas land plants require four enzymes: PDS (CrtP), ζ-carotene desaturase (ZDS, CrtQ), ζ-carotene isomerase (Z-ISO) and cis-carotene isomerase (CrtISO, CrtH). The gene encoding Z-ISO has been functionally identified in only two species, Arabidopsis thaliana and Zea mays, and has been little studied in other organisms. In this study, we found that the deduced amino acid sequences of Arthrospira Z-ISO and Euglena Z-ISO have 58% and 62% identity, respectively, with functional Z-ISO from Arabidopsis. We studied the function of Z-ISO genes from the cyanobacterium Arthrospira platensis and eukaryotic microalga Euglena gracilis. The Z-ISO genes of Arthrospira and Euglena were transformed into Escherichia coli strains that produced mainly 9,15,9′-tri-cis-ζ-carotene in darkness. In the resulting E. coli transformants cultured under darkness, 9,9′-di-cis-ζ-carotene was accumulated predominantly as Z-ISO in Arabidopsis. This indicates that the Z-ISO genes were involved in the isomerization of 9,15,9′-tri-cis-ζ-carotene to 9,9′-di-cis-ζ-carotene in darkness. This is the first functional analysis of Z-ISO as a ζ-carotene isomerase in cyanobacteria and eukaryotic microalgae. Green sulfur bacteria and Chloracidobacterium also use CrtP, CrtQ and CrtH for lycopene synthesis as cyanobacteria, but their genomes did not comprise Z-ISO genes. Consequently, Z-ISO is needed in oxygenic phototrophs, whereas it is not found in anoxygenic species.

The genus Arthrospira is filamentous, non-nitrogen-fixing cyanobacteria that is commercially important. We identified the molecular structures of carotenoids in Arthrospira platensis NIES-39. The major carotenoid identified was β-carotene. In addition, the hydroxyl derivatives of β-cryptoxanthin and (3R,3′R)-zeaxanthin were also found to be present. The carotenoid glycosides were identified as (3R,2′S)-myxol 2′-methylpentoside and oscillol 2,2′-dimethylpentoside. The methylpentoside moiety was a mixture of fucoside and chinovoside in an approximate ratio of 1 : 4. Trace amounts of the ketocarotenoid 3′-hydroxyechinenone were also found. Three types of lycopene cyclases have been functionally confirmed in carotenogenesis organisms. In cyanobacteria, the functional lycopene cyclases (CrtL, CruA and CruP) have only been found in four species. In this study, we found that CruA exhibited lycopene cyclase activity in transformed Escherichia coli, which contains lycopene, but CruP exhibited no lycopene cyclase activity and crtL was absent. This is the third cyanobacterial species in which CruA activity has been confirmed. Neurosporene was not a substrate of CruA in E. coli, whereas lycopene cyclases of CrtY (bacteria), CrtL (plants) and CrtYB (fungi) have been reported to convert neurosporene to 7,8-dihydro-β-carotene. β-Carotene hydroxylase (CrtR) was found to convert β-carotene to zeaxanthin in transformed E. coli, which contains β-carotene. Among the β-carotene hydroxylases, bacterial CrtZ and eukaryotic CrtR and BCH have similarities, whereas cyanobacterial CrtR appears to belong to another clade. Based on the identification of the carotenoids and the completion of the entire nucleotide sequence of the A. platensis NIES-39 genome, we propose a biosynthetic pathway for the carotenoids as well as the corresponding genes and enzymes.