Whey is a byproduct in the manufacturing of dairy products. Its use is limited, and its nutrients make it expensive for its disposal. Therefore, we attempted to develop its upcycling method. When the Aspergillus oryzae R...Whey is a byproduct in the manufacturing of dairy products. Its use is limited, and its nutrients make it expensive for its disposal. Therefore, we attempted to develop its upcycling method. When the Aspergillus oryzae RIB40 strain was cultured in whey, more than 75% of lactose, its major saccharide, remained after 7 days of cultivation. However, when lactase preparation was added, most sugars were efficiently removed, implying that lactase is essential for consuming whey nutrients. We created an A. oryzae lactase-overexpressing strain by introducing the tef1 promoter and lactase gene into RIB40. When cultured in whey, the transformant completely consumed its saccharides. The biological oxygen demand, chemical oxygen demand, total nitrogen concentration, and total phosphorus concentration of the whey decreased by 90%, 91%, 62%, and 95%, respectively, through this fermentation. The transformant can be considered "self-cloning" and is not subject to regulations on recombinant organisms in countries including Japan.
Akkermansia muciniphila, a mucin-degrading gut bacterium, contributes to intestinal homeostasis and metabolic disorders, yet its transcriptional response to human colonic mucin remains unclear. Here, we used human coloni...Akkermansia muciniphila, a mucin-degrading gut bacterium, contributes to intestinal homeostasis and metabolic disorders, yet its transcriptional response to human colonic mucin remains unclear. Here, we used human colonic organoids derived from either a healthy tissue (wild-type) or ATOH1-knockout lines lacking goblet cell differentiation. Co-culturing wild-type or ATOH1-knockout organoids with A. muciniphila followed by RNA-sequencing revealed distinct transcriptional profiles modulated by mucin availability, particularly affecting genes for growth and stress resistance. Notably, mucin degradation genes exhibited limited responses, contrasting with studies using porcine mucin, highlighting the specificity of human mucin interactions. Conversely, genes for mucin uptake and pili formation (e.g. Amuc_1100), which are crucial for host interaction, were upregulated with wild-type organoids. These results underscore the importance of using physiologically relevant human models. Our findings reveal A. muciniphila's adaptive gene regulation in response to human mucin, offering insights into host-microbe interactions shaped by the mucosal environment.
This study investigates the regulatory role of transmembrane protein TMEM164 in ferroptosis and autophagy in non-small cell lung cancer (NSCLC) cells, as well as its interaction with TRIM59. Gene expression analysis was...This study investigates the regulatory role of transmembrane protein TMEM164 in ferroptosis and autophagy in non-small cell lung cancer (NSCLC) cells, as well as its interaction with TRIM59. Gene expression analysis was conducted on NSCLC samples, and the effects of TMEM164 knockdown on ferroptosis and autophagy were examined in A549 cells. TMEM164 knockdown in A549 cells reduced ferroptosis by lowering lipid peroxidation and increasing cell viability, suggesting enhanced ferroptosis resistance. TRIM59 promoted ubiquitination and degradation of TMEM164, affecting autophagy and ferroptosis processes. Furthermore, TRIM59 knockdown reversed TMEM164's inhibition of autophagy-dependent ferroptosis, evidenced by changes in Fe2+, MDA, and GSH levels, as well as autophagy and ferroptosis-related protein expressions. Together, TMEM164 and TRIM59 play opposing roles in regulating autophagy and ferroptosis in NSCLC cells. TRIM59 knockdown inhibits the ubiquitination of TMEM164 to induce ferroptosis in NSCLC. This study offers insights for novel NSCLC treatment strategies.
A streamlined synthesis of sialylgalactose (NeuGal) analogs is described. Neuα(2,6)Gal and Neuα(2,3)Gal units were synthesized via fully α-selective sialylation of suitable Gal acceptors using a bicyclic sialyl donor. Ne...A streamlined synthesis of sialylgalactose (NeuGal) analogs is described. Neuα(2,6)Gal and Neuα(2,3)Gal units were synthesized via fully α-selective sialylation of suitable Gal acceptors using a bicyclic sialyl donor. NeuGals were diversified through C5 amino group modification and sulfation, yielding NeuGal analogs with an amino linker for biological studies.
Renal fibrosis is a pathological feature of chronic kidney injury that contributes to renal failure. This study aimed to explore the effects of Hirudin on renal fibrosis. The antifibrotic effect of Hirudin was evaluated...Renal fibrosis is a pathological feature of chronic kidney injury that contributes to renal failure. This study aimed to explore the effects of Hirudin on renal fibrosis. The antifibrotic effect of Hirudin was evaluated using unilateral ureteral obstruction (UUO) rats and TGF-β-treated HK-2 cells. The autophagy inhibitor 3-methyladenine was used to further explore the potential mechanism. Hirudin treatment significantly reduced UUO-induced elevations in blood urea nitrogen, creatinine, and alpha-smooth muscle actin (α-SMA) levels and improved kidney injury and renal fibrosis. In addition, Hirudin markedly decreased NLRP3 inflammasome-related protein expression and increased autophagy-related protein expression in the kidneys of UUO rats. Hirudin significantly increased cell viability, reduced α-SMA and NLRP3 inflammasome-related protein levels, and increased autophagy-related protein levels in TGF-β-treated HK-2 cells. However, the effects of Hirudin were counteracted by 3-methyladenine. In conclusion, Hirudin inhibits the activation of NLRP3 inflammasome by inducing autophagy to improve renal fibrosis.
β-Glucosidase converts soybean isoflavone glucosides to aglycones, and Bacillus subtilis strain Miyagino shows remarkably lower activity than B. subtilis strain 168, when cultured in a soybean-based medium. The inactivat...β-Glucosidase converts soybean isoflavone glucosides to aglycones, and Bacillus subtilis strain Miyagino shows remarkably lower activity than B. subtilis strain 168, when cultured in a soybean-based medium. The inactivation of bglH, a key β-glucosidase-related gene, was found in B. subtilis strain Miyagino. This study indicates that the decreased bglH expression significantly affects the low β-glucosidase activity of B. subtilis strain Miyagino.
This study presents the first detailed evidence that commercially available kuzu-mochi products-a traditional Japanese sweet made from fermented wheat starch-are a natural source of vitamin B12 compounds. Microbiological...This study presents the first detailed evidence that commercially available kuzu-mochi products-a traditional Japanese sweet made from fermented wheat starch-are a natural source of vitamin B12 compounds. Microbiological assays and liquid chromatography-tandem mass spectrometry analyses confirmed the presence of vitamin B12 and its analog (pseudovitamin B12) across all tested samples, along with minor corrinoid compounds. Significant variability in vitamin B12 content was noted at approximately 25-152 ng/100 g wet weight, which was probably associated with differences in fermentation-derived microbial communities. Although kuzu-mochi cannot be a primary source of vitamin B12, its role as a supplementary dietary source is remarkable. These findings shed new light on the nutritional potential of traditional fermented foods.
The metal-reducing bacterium Geobacter sulfurreducens PCA is capable of anaerobic respiration using elemental sulfur as an electron acceptor. Despite 3 decades since its isolation, the molecular mechanisms underlying sul...The metal-reducing bacterium Geobacter sulfurreducens PCA is capable of anaerobic respiration using elemental sulfur as an electron acceptor. Despite 3 decades since its isolation, the molecular mechanisms underlying sulfur respiration remain unclear. In this study, we conducted a transcriptome analysis of G. sulfurreducens PCA cultured with and without sublimed sulfur. In the presence of sulfur, 153 genes were significantly up-regulated, while 599 genes were down-regulated. Notably, genes encoding redox proteins involved in energy conservation, particularly multiheme c-type cytochromes, exhibited altered expression patterns. In addition, elemental sulfur induced the transcription of genes associated with sulfur, selenium, and nitrogen metabolism, as well as protein redox homeostasis, DNA repair, and even cell motility. These transcriptional responses may reflect metabolic adaptation to sulfur respiration, redox state alterations, and sulfur-induced stress. Our findings uncover a complex regulatory landscape governing sulfur respiration and provide critical insights into this long-standing biochemical enigma in G. sulfurreducens PCA.
In vascular endothelial cells, proinflammatory cytokines and Toll-like receptor (TLR) ligands up-regulate the expression of adhesion molecules. In the present study, screening of the RIKEN Natural Products Depository che...In vascular endothelial cells, proinflammatory cytokines and Toll-like receptor (TLR) ligands up-regulate the expression of adhesion molecules. In the present study, screening of the RIKEN Natural Products Depository chemical libraries identified podophyllotoxin and α-peltatin, which inhibited intercellular adhesion molecule-1 (ICAM-1) expression induced by polyinosinic-polycytidylic acid (Poly(I:C)) as a TLR3 ligand. In human umbilical vein endothelial cells (HUVEC), podophyllotoxin and its derivative α-peltatin inhibited Poly(I:C)-induced increases in the mRNA expression of ICAM-1, vascular cell adhesion molecule-1 (VCAM-1), and E-selectin. In addition, podophyllotoxin and α-peltatin inhibited the Poly(I:C)-induced nuclear translocation of the nuclear factor κB (NF-κB) subunit RelA. Microtubule-targeting agents (colchicine, vinblastine, and paclitaxel) exerted different effects on the Poly(I:C)-induced mRNA expression of ICAM-1, VCAM-1, and E-selectin. Vinblastine potently inhibited nuclear RelA translocation in Poly(I:C)-stimulated HUVEC, whereas colchicine and paclitaxel did not. Collectively, these results demonstrate that podophyllotoxin and α-peltatin inhibited NF-κB activation and mRNA expression by TLR3 stimulation in HUVEC.
Acidic polysaccharides such as alginate, a key component of brown algae, have unique properties conferred by their carboxyl groups. Alginate is degraded by alginate lyases, a class of polysaccharide lyases (PLs) that cle...Acidic polysaccharides such as alginate, a key component of brown algae, have unique properties conferred by their carboxyl groups. Alginate is degraded by alginate lyases, a class of polysaccharide lyases (PLs) that cleave uronic acid glycoside bonds via β-elimination. These enzymes, which are classified into various PL families, differ in structure and substrate specificity but frequently share structural motifs including β-helices, β-jelly rolls, and (α/α)6 barrels coupled with antiparallel β-sheets. Moreover, marine bacteria from the genera Alteromonas, Pseudoalteromonas, and Vibrio produce alginate lyases that belong to several PL families that are associated with blue carbon cycling. Furthermore, some Bacteroides species in the human gut have acquired alginate-degrading genes via horizontal transfer from marine bacteria and/or other Bacteroides species. Alginate fermentation by gut microbes can produce short-chain fatty acids with potential prebiotic effects. This review explores alginate lyase diversity, ecological roles, and relevance in both marine and human microbial ecosystems.
Efficient intracellular delivery of functional proteins into the yeast Saccharomyces cerevisiae remains a major technical challenge due to its rigid cell wall. Here, we report that synthetic peptide tags, particularly pe...Efficient intracellular delivery of functional proteins into the yeast Saccharomyces cerevisiae remains a major technical challenge due to its rigid cell wall. Here, we report that synthetic peptide tags, particularly penetratin, enable nearly complete delivery of GFP, albeit with substantial cytotoxicity. This method provides a promising non-genetic platform for the intracellular delivery and functional analysis of proteins in yeast systems.
Coelenterazine (CTZ) and dehydrocoelenterazine (dCTZ, a dehydrogenated form of CTZ) were first identified in the liver of the luminescent squid Watasenia scintillans. In this report, we demonstrate for the first time tha...Coelenterazine (CTZ) and dehydrocoelenterazine (dCTZ, a dehydrogenated form of CTZ) were first identified in the liver of the luminescent squid Watasenia scintillans. In this report, we demonstrate for the first time that CTZ can be successfully converted to dCTZ using molecular sieves (MS4A/Na+, MS13X/Na+, MS5A/Ca2+, and MS3A/K+) in isopropyl alcohol at 37 °C for 3 h. The conversion efficiency to dCTZ followed the order: MS4A/Na+ = MS13X/Na+ > MS5A/Ca2+ > MS3A/K+. Lower efficiencies observed with modified MS4A (TMS-MS4A, NH4+-MS4A, and H+-MS4A) suggest that cations in the molecular sieves play a crucial role in the dehydrogenation reaction of CTZ. Notably, MS4A recovered from the reaction mixture could be reused without thermal regeneration for dehydration. In purified dCTZ, no residual CTZ was detected using the luciferin-luciferase reaction, and dCTZ can be used as a substrate in luminescence assays to determine enzymatic conversion activity to CTZ.
Endothelial-to-mesenchymal transition (EndMT) is a process that causes endothelial cells (ECs) to lose their EC characteristics and transform into mesenchymal cells. Accumulating studies suggest that EndMT is induced in...Endothelial-to-mesenchymal transition (EndMT) is a process that causes endothelial cells (ECs) to lose their EC characteristics and transform into mesenchymal cells. Accumulating studies suggest that EndMT is induced in atherosclerotic plaques and contributes to the pathogenesis of atherosclerosis. LL-37 is a multifaceted peptide with antimicrobial and immunomodulatory actions. Interestingly, LL-37 is localized in atherosclerotic plaques, suggesting an association between EndMT and LL-37 in atherosclerosis. Thus, we examined the EndMT-inducing activity of LL-37 using human umbilical vein ECs. LL-37 decreased EC markers but increased mesenchymal cell markers in the cells. LL-37 decreased the vascular network formation of the cells but increased the cell migration, a characteristic function of mesenchymal cells. Finally, the LL-37-induced EndMT was inhibited by Akt and nuclear factor-kappa B (NF-κB) inhibitors, suggesting that LL-37 induces EndMT by activating Akt and NF-κB. These observations speculate a role of LL-37 in the pathogenesis of atherosclerosis as an EndMT inducer.
The transceptor Can1 negatively regulates proline utilization in the yeast Saccharomyces cerevisiae. Here, we demonstrated that Can1 physically interacts with the catalytic subunits of protein kinase A (Tpk1, Tpk2, and T...The transceptor Can1 negatively regulates proline utilization in the yeast Saccharomyces cerevisiae. Here, we demonstrated that Can1 physically interacts with the catalytic subunits of protein kinase A (Tpk1, Tpk2, and Tpk3). Furthermore, we identified a specific site in Can1 that is essential for inhibiting proline utilization. These findings provide a mechanistic basis for Can1-mediated metabolic regulation.
We developed an electroporation-based transformation system for Moniliella megachiliensis, a basidiomycetous yeast producing erythritol. Using a green fluorescence protein expression vector, successful gene transfer and...We developed an electroporation-based transformation system for Moniliella megachiliensis, a basidiomycetous yeast producing erythritol. Using a green fluorescence protein expression vector, successful gene transfer and expression were confirmed. This is the first report of genetic transformation in this genus, providing a foundation for future genetic engineering and biotechnological applications in food and fermentation industries.
Labor shortages threaten global apple production, thereby encouraging new strategies to improve orchard management. The growth of columnar apples, controlled by the MdDOX-Co gene, enables vertical growth with minimal lat...Labor shortages threaten global apple production, thereby encouraging new strategies to improve orchard management. The growth of columnar apples, controlled by the MdDOX-Co gene, enables vertical growth with minimal lateral branching, allowing for high-density planting and easier harvesting. MdDOX-Co encodes 2-oxoglutarate-dependent dioxygenase (2ODD, DOX). This study aimed to identify selective chemical inhibitors of MdDOX-Co. We synthesized the parental C6-based analogs featuring a heterocyclic 1,3,4-oxathiazol-2-one ring and evaluated their inhibitory activity. Compounds retaining the 1,3,4-oxathiazol-2-one core exhibited strong in vitro inhibition and promoted seedling elongation in MdDOX-Co overexpressing Arabidopsis. Structure-activity analysis confirmed that the 1,3,4-oxathiazol-2-one ring was essential, with tolerance for side-chain variations, including bulky groups. Selectivity assays indicated minimal off-target effects on the related 2ODD enzymes. Molecular modeling suggested the compatibility of the lead compounds with the MdDOX-Co active site. These findings encourage us to develop MdDOX-Co-targeted agrochemicals to chemically regulate tree architecture and enhance productivity during apple cultivation.
Fission yeast spores possess strong resistance to environmental stresses, largely due to the outermost proteinaceous "Isp3 layer," which comprises Isp3 protein. Isp3 is palmitoylated, and its localization to the spore pe...Fission yeast spores possess strong resistance to environmental stresses, largely due to the outermost proteinaceous "Isp3 layer," which comprises Isp3 protein. Isp3 is palmitoylated, and its localization to the spore periphery is impaired in mutants lacking palmitoyltransferase; however, the precise role of Isp3 palmitoylation remains unclear. Here, we found that Isp3-GFP was expressed at wild-type levels in forming spores in mug142∆ cells lacking the palmitoyltransferase catalytic unit; thus, lack of palmitoylation did not reduce Isp3 protein stability. Next, we identified cysteine 7 as the key palmitoylation site essential for Isp3 localization to the spore periphery. Electron microscopy revealed that the Isp3 fibrillar layer was absent in both mug142Δ and isp3-C7S spores. Additionally, the isp3-C7S spores displayed increased sensitivity to alcohol stress, similar to isp3∆ spores. Collectively, these results demonstrate that palmitoylation of Isp3 is essential for the relocation of Isp3 to the spore surface and the assembly of the Isp3 layer.
Escherichia coli TrkG and TrkH transporters contain a unique N-terminal Domain-0 (D0). Our findings reveal that D0 supports both the function and stability of TrkG, enabling K+ and Na+ uptake, whereas it is not essential...Escherichia coli TrkG and TrkH transporters contain a unique N-terminal Domain-0 (D0). Our findings reveal that D0 supports both the function and stability of TrkG, enabling K+ and Na+ uptake, whereas it is not essential for TrkH-mediated K+ uptake. This difference can be attributed to D0 role in stabilizing polar residues within TrkG core transmembrane domains.
Listeria monocytogenes, which grows in host cells intracellularly, is important as an opportunistic pathogen. Extracts of Chaga mushroom, Inonotus obliquus, have been shown to possess various biological activities. This...Listeria monocytogenes, which grows in host cells intracellularly, is important as an opportunistic pathogen. Extracts of Chaga mushroom, Inonotus obliquus, have been shown to possess various biological activities. This study explores the impact of Chaga extracts on host defense against infection with L. monocytogenes, which induces T-helper (Th)1-type immune response. Chaga extracts enhanced cytokine production including tumor necrosis factor-α, interleukin (IL-6), and IL-12 but suppressed IL-10 production in murine macrophages and dendritic cells. Oral administration of Chaga extracts inhibited the proliferation of L. monocytogenes in the livers of infected mice. Interferon-γ (IFN-γ) and IL-12 production was enhanced in the spleen cell cultures of the extract-treated mice. Chaga extract treatment augmented the clearance of L. monocytogenes from the livers and spleens during re-infection. IFN-γ production was also enhanced in the spleen cell cultures of primarily infected mice. Together, the results indicate that Chaga extracts possess protective potentials for infectious diseases such as listeriosis.