α-1,3-Glucanase from Streptomyces thermodiastaticus HF3-3 (Agl-ST) has been classified in the glycoside hydrolase (GH) family 87. Agl-ST is a multi-modular domain consisting of an N-terminal β-sandwich domain (β-SW), a catalytic domain, an uncharacterized domain (UC), and a C-terminal discoidin domain (DS). Although Agl-ST did not hydrolyze α-1,4-glycosidic bonds, its amino acid sequence is more similar to GH87 mycodextranase than to α-1,3-glucanase. It might be categorized into a new subfamily of GH87. In this study, we investigated the function of the domains. Several fusion proteins of domains with green fluorescence protein (GFP) were constructed to clarify the function of each domain. The results showed that β-SW and DS domains played a role in binding α-1,3-glucan and enhancing the hydrolysis of α-1,3-glucan. The binding domains, β-SW and DS, also showed binding activity toward xylan, although it was lower than that for α-1,3-glucan. The combination of β-SW and DS domains demonstrated high binding and hydrolysis activities of Agl-ST toward α-1,3-glucan, whereas the catalytic domain showed only a catalytic function. The binding domains also achieved effective binding and hydrolysis of α-1,3-glucan in the cell wall complex of Schizophyllum commune.

Androgenetic alopecia (AGA) involves dihydrotestosterone-driven follicular miniaturization compounded by oxidative stress and perifollicular inflammation, while current pharmacotherapies remain limited by adverse effects. Neem (Azadirachta indica A. Juss., Meliaceae) leaves, a phenolic- and limonoid-rich plant widely used in Southeast Asian traditional medicine, were fermented for seven days with a tri-culture consortium of Saccharomyces cerevisiae, Lactobacillus plantarum, and Aspergillus niger (TRI-NE) to evaluate whether expanded microbial diversity enhances bioactivity relative to the unfermented extract (UN-NE). TRI-NE significantly increased total phenolic content, rising from 315.30 to 564.70 mg GAE/g by day 7, alongside an overall enhancement of antioxidant capacity across all assays. Moreover, untargeted metabolomics revealed compositional remodeling, with 67.47% of significantly altered metabolite features upregulated after fermentation, including selective enrichment of quercetin despite reductions in other polyphenols. In human hair follicle dermal papilla cells (HFDPCs) and complementary models, TRI-NE consistently outperformed UN-NE, enhancing paracrine-mediated fibroblast proliferation, preserving cell viability under potassium-channel blockade, suppressing lipopolysaccharide-induced inflammatory nitric oxide production, and attenuating oxidative membrane damage. At the transcriptional level, TRI-NE downregulated androgen metabolism (SRD5A1 and SRD5A2) and pro-regression genes (TGFB1) while upregulating Wnt/β-catenin (CTNNB1), Sonic Hedgehog (SHH, SMO, and GLI1), and angiogenic (VEGF) pathway genes, with effects matching or exceeding standard hair-loss therapeutics. These findings indicate that TRI-NE confers superior bioactivity over the unfermented extract, supporting its potential as a multi-target cosmeceutical candidate for AGA.

Androgenetic alopecia (AGA), the most common form of hair loss, is driven by dihydrotestosterone-mediated follicular miniaturization, oxidative stress, and perifollicular inflammation, requiring multi-target intervention. Coffee pulp is an abundant coffee-processing by-product and a potential source of value-added bioactives relevant to AGA. This study investigated whether fermentation with Saccharomyces cerevisiae, Lactobacillus plantarum, or their mixed culture could remodel coffee pulp composition and enhance biological activities relevant to AGA. Untargeted metabolomics revealed treatment-dependent metabolic remodeling. LP-CP showed the largest number of significantly altered metabolite features (1020; 666 increased and 354 decreased), whereas MIX-CP exhibited a predominantly upward pattern, with 238 of 283 significantly altered features (84.10%) showing increased abundance relative to the unfermented control. Targeted polyphenol analysis showed MIX-CP contained the highest levels of rosmarinic acid and quercetin. In human hair follicle dermal papilla cells, MIX-CP stimulated proliferation, increased fibroblast proliferation in a conditioned-medium model, partially rescued cell viability under potassium-channel inhibition, and attenuated intracellular reactive oxygen species and lipid peroxidation more than monoculture-fermented extracts. At the transcriptional level, MIX-CP downregulated androgen metabolism genes SRD5A1 and SRD5A2 alongside pro-regression mediator TGFB1 while upregulating Wnt/β-catenin (CTNNB1), Sonic Hedgehog (SHH, SMO, GLI1), and angiogenic (VEGF) genes, showing transcript-level modulation comparable to standard hair-loss drugs for most targets. These findings position MIX-CP as a promising multi-target cosmeceutical for AGA management, warranting further in vivo validation.