Bioresources and Bioprocessing最新文献

{"title":"Harnessing spent granular activated carbon from point-of-use water treatment for anaerobic digestion enhancement: characterization and pretreatment effects.","authors":"J B Kemirembe, A Ayor, T Kayondo, R Kayiwa","doi":"10.1186/s40643-025-00947-9","DOIUrl":"https://doi.org/10.1186/s40643-025-00947-9","url":null,"abstract":"<p><p>As an alternative to environmentally unfriendly disposal of spent activated carbon (SAC) from point-of-use water treatment systems, this study harnessed SAC as a co-substrate in anaerobic digestion for biogas production. The study investigated how SAC pre-treatment methods, namely, washing, particle size (milled vs. granular), and dosing levels (2.5%, 5%, and 7.5%), influenced biogas yield, methane content, and retention time using cow dung as the primary substrate. SAC from chlorinated water purification (KC) and the other from borehole water treatment (KZ) were investigated in this study. KZ had a higher %TOC (5.4955%) compared to < LOD for KC, indicating it retained more usable carbon for microbial activity. However, KC had a higher surface area (110.58 m²/g) than KZ (78.41 m²/g). suggesting better microbial support. Digesters dosed with 2.5% and 5% SAC generally maintained the most stable retention time, sustaining active digestion across the full 30-day period. Overall, unwashed granular KZ dosages of 2.5%, 5.0%, and 7.5% respectively, supported the highest and most stable biogas output at 5047 mL, 2605 mL, and 1685 mL, respectively. A 2.5% KZ dosage outperformed the control (pure cow dung), which produced 2624 mL. Milled washed KC dosages of 2.5%, 5.0%, and 7.5% generally showed lower biogas outputs, yielding 217.8 mL, 540.8 mL, and 300 mL, respectively. Their relatively low performance suggests that even after washing, the milled coconut husk SAC, at these concentrations, did not significantly enhance microbial activity, or perhaps the fine particle size led to aggregation or hindered mass transfer. KZ-dosed digesters' highest methane composition range was 14.90-37.70% whereas for KZ digesters it was 47.73-52.53% compared to the control's 33.70%. These findings underscore the complex interplay between particle size, washing, and dosage of SAC for enhancing anaerobic digestion, necessitating optimization.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"111"},"PeriodicalIF":5.1,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145257264","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Agro-industrial by-products valorization for fructooligosaccharide production with Zymomonas mobilis.","authors":"Adelaide Braga, Ana Benedita Maia, Lígia R Rodrigues","doi":"10.1186/s40643-025-00887-4","DOIUrl":"10.1186/s40643-025-00887-4","url":null,"abstract":"<p><p>Fructooligosaccharides (FOS) have gained attention due to their prebiotic properties and potential health benefits. This study explores the production of FOS using Zymomonas mobilis ZM4, a promising candidate for biotechnological processes, utilizing corn steep liquor (CSL) and sugarcane molasses as alternative and sustainable carbon and nitrogen sources. Two distinct media formulations were investigated, namely one composed of CSL supplemented with yeast extract (YE), and another utilizing sugarcane molasses. CSL was evaluated at concentrations of 10 g L^-1 and 12 g L^-1 in combination with YE. The optimal combination, 12 g L^-1 CSL and 8 g L^-1 YE, yielded 60.00 ± 0.44 g L^-1 FOS with a productivity of 1.250 ± 0.009 g L^-1 h^-1, comparable to synthetic media. Molasses, another agro-industrial by-product, was tested at sucrose-equivalent concentrations of 150, 200, and 350 g L^-1. The highest FOS concentration, 58.67 ± 1.64 g L^-1, was achieved with 200 g L^-1 of molasses. Combining CSL and molasses (CSLM media) resulted in 58.15 ± 0.21 g L^-1 of FOS with a yield of 0.307 ± 0.003 g_FOS g_sucrose^-1. The FOS mixture included 1-kestose, 6-kestose, nystose, and neokestose. Although scaling up to a bioreactor led to a lower FOS concentration of 42.31 ± 0.16 g L^-1, the yield remained promising at 0.482 ± 0.008 g_FOS g_sucrose^-1. This study not only highlights the efficient production of FOS using Z. mobilis ZM4 but also demonstrates the potential of using CSL and molasses, byproducts of agro-industrial processes, as cost-effective and sustainable substrates for industrial-scale FOS production. The findings provide valuable insights for the development of bio-based processes for functional oligosaccharide production.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"110"},"PeriodicalIF":5.1,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12500493/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145238173","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving the energy balance between microwave pretreatment and anaerobic digestion with centrifuged biosludge.","authors":"Sukru Aslan, Ali Alhraishawi","doi":"10.1186/s40643-025-00903-7","DOIUrl":"10.1186/s40643-025-00903-7","url":null,"abstract":"<p><p>The effect of pretreatment of raw biosludge (Bs) and centrifuged Bs (CBs) with microwave (MW) operated at constant conditions before an anaerobic digestion (AD) process on the energy requirement and production of hybrid system was investigated. With the successful breakdown of Bs and CBs by the MW irradiation, the soluble COD concentrations increased from average value of about 76 mg/L to 300 mg/L and 6400 mg/L for the Bs and CBs, respectively. After the MW disintegration, the protein and sugar concentrations were about 10 and 22 times higher in the pretreated samples compared to the biosludges. While, the total and volatile solids (TS and VS) concentrations in the liquor declined. The increase of readily bioavailable organics in pretreated samples improved about 22% CH₄ production in AD units, compared to the raw Bs and CBs. It was determined that the energy requirement of MW irradiation decreased about 3 times with the centrifugation of Bs. Although the applied whole models were well correlation, the kinetic analysis showed that Transference Function was the best model with the highest correlation coefficient (R² between 0.953 and 0.998) for predicting the most accurately CH₄ production. The nutrients concentrations in effluent of AD process was significantly higher than the liquors obtained after the MW irradiation was observed.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"109"},"PeriodicalIF":5.1,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12491126/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145205233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Divergent immunomodulatory and gut microbiota-modulating effects of Sargassum polysaccharides and oligosaccharides in delayed-type hypersensitivity mice.","authors":"Yang-Ching Chen, Shih-Yuan Fang, Chien-Li Chen, Ming-Chih Fang, Yu-Ying Yang, Meng-Chou Lee, Chung-Hsiung Huang","doi":"10.1186/s40643-025-00948-8","DOIUrl":"10.1186/s40643-025-00948-8","url":null,"abstract":"<p><p>Recent research has indicated that polysaccharides extracted from Sargassum (SP) possess promising activity in alleviating type I hypersensitivity reactions. However, effects of SP and Sargassum oligosaccharides (SO) on immune regulation and gut microbiota in type IV hypersensitivity remain unexplored. In this study, SP and SO were prepared and structurally characterized. SP contained high-molecular-weight fractions (866 kDa and 276 kDa), whereas SO was composed of low-molecular-weight components (3.74 kDa and 126 Da), lacked sulfate groups, and exhibited higher reducing sugar contents. The influence of SP and SO on immune regulation and the structure of gut microbial communities was examined using a mouse model of ovalbumin (OVA)-induced delayed-type hypersensitivity (DTH). Administration of SP and SO (25 or 250 mg/kg per day for 10 days) significantly attenuated DTH responses, evidenced by a decrease in footpad edema and a lower degree of cell infiltration. While both SP and SO had limited effects on serum IgG₁ levels and splenic TGF-β production, treatment with SO at 250 mg/kg significantly reduced serum total IgG, OVA-specific IgG, and splenic IL-2 levels, while increasing IL-10 production. Notably, SO exerted the most pronounced effect in lowering IgG_2a and IFN-γ levels. Additionally, SO treatment led to distinct shifts in gut microbial profile, marked by elevated levels of Muribaculaceae, Lactobacillus, and Bacteroides. These microbial changes were accompanied by elevated concentrations of short-chain fatty acids (SCFAs). Collectively, these results indicate that SO holds potential as a functional dietary component for the alleviation of type IV hypersensitivity responses, through modulation of gut microbiota and immunomodulation.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"108"},"PeriodicalIF":5.1,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12484449/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145197627","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enzymatic hydrolysis of almond hulls for cultivation of edible filamentous fungi.","authors":"Lin Cao, Allan Chio, Hamed M El Mashad, Zhongli Pan, Ruihong Zhang","doi":"10.1186/s40643-025-00940-2","DOIUrl":"10.1186/s40643-025-00940-2","url":null,"abstract":"<p><p>Enzymatic hydrolysis and liquefaction of the residual almond hull solids (RAHS) after sugar extraction were studied. The hydrolysis performance of three different enzymes (Cellic CTec2, Viscozyme L and Pectinex Ultra SPL) were investigated. The ability of Aspergillus awamori and Aspergillus oryzae in utilizing the hydrolysate was evaluated. In 50 mL Falcon tubes, the combination of 200 [Formula: see text]L/g RAHS of Cellic Ctec2 and 60 [Formula: see text]L/g RAHS of Viscozyme L enzymes resulted in the optimal performance, with total sugar yield of 47.21%, total fiber conversion of 86.01%, and liquefaction efficiency of 51.61%. Applying these optimal conditions in 250 mL glass bottles resulted in higher liquefaction efficiency of 72.53%, while total fiber conversion was same as that in Falcon tubes. Through a combination of sugar extraction and subsequent enzymatic hydrolysis, a total of 0.55 g of sugars could be released per gram of almond hulls (dry basis, d.b.). The cultivation of Aspergillus awamori and Aspergillus oryzae in hydrolysate for five days produced uniform fungal pellets, with biomass yields of 0.89 g total suspended solids (TSS)/g sugar and 0.43 g TSS/g sugar, respectively. Both fungal strains were able to utilize sugars in the hydrolysate, including galacturonic acid. This research demonstrates that enzymatic hydrolysis of RAHS enables the recovery of additional sugars from almond hulls, thereby increasing the yield of fungal biomass and improving the overall utilization of almond hulls.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"107"},"PeriodicalIF":5.1,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12477097/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145184608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an in vitro multi-enzyme system for efficient one-pot biosynthesis of sorbitol from fructose-6-phosphate.","authors":"Kai Shen, Chao-Nan Zhu, Jian-He Xu, Gao-Wei Zheng, Qi Chen","doi":"10.1186/s40643-025-00943-z","DOIUrl":"10.1186/s40643-025-00943-z","url":null,"abstract":"<p><p>Fructose-6-phosphate is a key ketose monophosphate intermediate that plays a central role in the metabolic pathway of glycolysis. Sorbitol is a naturally occurring sugar alcohol with diverse biological properties, which is widely applied in the food, cosmetic, and pharmaceutical industries due to its favorable physicochemical and physiological characteristics. Although a biosynthetic pathway from starch to fructose-6-phosphate has been well established in metabolic engineering studies, the downstream enzymatic route leading to the biosynthesis of sorbitol from this intermediate has not yet been elucidated in the literature. In this study, we performed the design and implementation of an artificial multi-enzymatic cascade system for the biosynthesis of sorbitol from fructose-6-phosphate. Firstly, candidate enzymes sorbitol-6-phosphate dehydrogenase (S6PDH) and sorbitol-6-phosphate dephosphatase (S6PDP) were selected with an initial activity of 147 U/mg and 1.7 U/mg, respectively. Secondly, molecular modifications were then carried out to improve the thermostability of EcS6PDH, EcS6PDH-M4 displays markedly improved thermostability, with a significant extension of half-life at 40 °C from less than 1 min to 375 min and an elevated T_m by 9.1 °C. Although multiple rounds of protein engineering were undertaken to improve the catalytic activity of EcS6PDP, no variant exhibiting substantially enhanced catalytic efficiency was identified. Finally, a one-pot biosynthetic system was established by integrating EcS6PDH-M4, EcS6PDP, and glucose dehydrogenase. Following systematic optimization of this multi-enzyme cascade system, 82.6 mM of sorbitol was efficiently produced from 200 mM fructose-6-phosphate. Our work provides a foundational framework for the further enzymatic synthesis of sorbitol directly from starch, offering a promising route for sustainable sugar alcohol production.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"104"},"PeriodicalIF":5.1,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12474828/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145147727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of fermentation parameters, volatile compounds and microbial community structure of fermented grains in the upper and lower layers during strong-flavor Baijiu fermentation.","authors":"Jie Deng, Chao-Jiu He, Chun-Hui Wei, Hui-Bo Luo, Zhi-Guo Huang","doi":"10.1186/s40643-025-00916-2","DOIUrl":"10.1186/s40643-025-00916-2","url":null,"abstract":"<p><p>Strong-flavor Baijiu (SFB) is produced using a solid-state fermentation system, but the impact of stratified fermented grains on fermentation remains unclear. Therefore, in a typical distillery plant, we evaluated the physical and chemical composition, volatile compound profile, and microbial community of fermented grains in SFB both above (FG-A) and below (FG-B) the Huangshui line. Significant differences in fermentation parameters between FG-A and FG-B were observed after 30 days of fermentation (P < 0.05). Additionally, the partial least squares projection to latent structure discriminant analysis (PLS-DA) revealed distinct differences in volatile compounds between FG-B and FG-A, identifying 26 discriminant markers. The diversity of short-chain fatty acids (SCFAs) and their esters were higher in FG-B compared to FG-A. Furthermore, microbial diversity and abundance have differed significantly between the two layers of fermented grains (P < 0.05), included 17 differential genera. Correlation and pathway enrichment analyses indicated that the higher SCFA content in FG-B could be attributed to the greater abundance of acid-producing microorganisms compared to FG-A. This study highlights the differences between the two layers of fermented grains in SFB fermentation, offering new insights into solid-state fermentation and expanding the current understanding of the traditional SFB fermentation process.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"105"},"PeriodicalIF":5.1,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12474805/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145147664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fruiting body-associated Pseudomonas contact triggers ROS-mediated perylenequinone biosynthesis in Shiraia mycelium culture.","authors":"Yan Jun Ma, Xin Ping Li, Jia Hui Li, Li Ping Zheng, Jian Wen Wang","doi":"10.1186/s40643-025-00946-w","DOIUrl":"10.1186/s40643-025-00946-w","url":null,"abstract":"<p><p>Perylenequinones (PQs) from Shiraia fruiting bodies serve as potent photosensitizers for anticancer and antimicrobial photodynamic therapy (PDT). Although these fruiting bodies harbor diverse endophytic bacteria, their interactions with the host fungus remain poorly understood. In this study, we used an in vitro confrontation bioassay to investigate the interaction between Shiraia sp. S9 and dominant Pseudomonas isolates, analyzing fungal transcriptional responses and PQ biosynthesis. Comparative assessment of co-cultures with freely suspended live P. fulva SB1 versus dialysis membrane-separated bacteria revealed that direct physical contact is essential for eliciting fungal PQ production, particularly extracellular secretion of hypocrellin A (HA), HC, and elsinochrome A-C. Bacterial elicitation with P. fulva SB1 at 400 cells/mL stimulated both intracellular PQ biosynthesis and extracellular secretion, resulting in a total PQ yield of 362.2 mg/L, a 2.4-fold increase over axenic cultures. RNA-seq analysis after 24 h of co-culture identified 646 differentially expressed genes (DEGs), with 445 upregulated and 201 downregulated, showing significant enrichment in oxidative stress defense, carbohydrate metabolism, and membrane transport functions. Bacterial contact induced reactive oxygen species (ROS) generation, specifically O₂^·- and H₂O₂, which mediated increased membrane permeability and enhanced HA production. This was achieved through upregulation of key genes involved in central carbon metabolism, polyketide synthase (PKS) for PQ biosynthesis, and major facilitator superfamily (MFS) transporter for PQ exudation. Our work provides the first evidence that the contact-dependent ROS signaling by endophytes within fruiting bodies regulates fungal secondary metabolism, offering novel insights into bacterial-fungal interactions and establishing an effective co-culture strategy for enhanced production of bioactive PQs.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"106"},"PeriodicalIF":5.1,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12474784/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145147724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancements in the application of reporter gene cell lines in bioactivity evaluation of biological products.","authors":"Kaijuan Yi, Can Wang, Huili Lu","doi":"10.1186/s40643-025-00932-2","DOIUrl":"10.1186/s40643-025-00932-2","url":null,"abstract":"<p><p>The assessment of biological product activity is a key aspect of quality control. Currently, in vitro assays serve as the primarily method employed by both companies and regulatory agencies to evaluate biological activity. Reporter Gene Assay (RGA) is a technique that investigates gene expression regulation and cellular signal transduction pathway activation through easily detectable reporter genes. RGA is highly dependent on drug mechanisms, offering high accuracy and precision, and has gained increasing recognition. The utilization of alternative analytical methods based on RGA have emerged as a prevailing trend, with a growing number of antibody drugs adopting corresponding RGA-based quality control approaches. Establishing stable expressing cell lines is essential to ensure the stability, reliability, and consistency of assays across diverse conditions when employing RGA techniques. CRISPR/Cas9 gene editing technology mediated site-specific gene integration allows for rapid and precise insertion of exogenous genes into specific genomic loci and enables the efficient construction of stable RGA cell lines, which would significantly propel the advancement of biological activity evaluation methods.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"103"},"PeriodicalIF":5.1,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12460214/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145136419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring green proteins from pumpkin leaf biomass: assessing their potential as a novel alternative protein source and functional alterations via pH-Shift treatment.","authors":"Marija Korićanac, Jelena Mijalković, Predrag Petrović, Neda Pavlović, Zorica Knežević-Jugović","doi":"10.1186/s40643-025-00945-x","DOIUrl":"10.1186/s40643-025-00945-x","url":null,"abstract":"<p><p>As sustainability continues to be a major global focus, the use of agro-industrial waste as a renewable resource presents a promising solution. In this study, the potential of pumpkin leaf biomass, a type of agro-industrial waste, will be explored as an alternative protein source using thermal coagulation. A key challenge is the accumulation of green proteins, a protein-rich byproduct. This waste fraction, which has been insufficiently investigated due to its limited functional properties, is the main focus of this research, aimed at enabling complete biomass utilization. The novelty of this research lies in the application of combined pH-shift and controlled heat treatment to improve the solubility of green proteins isolated from pumpkin leaf biomass - an underexplored protein fraction derived from agro-industrial waste. The obtained crude green protein powder, with a yield of 47.95 g/kg of leaf dry biomass, exhibited a satisfactory composition, containing 53.58% protein, a high-quality amino acid profile, and notable antioxidant properties. On the other hand, it exhibited low solubility, below 25%, across the pH range of 2-10, with the isoelectric point at pH 4.4. Following the application of an optimized pH-shift and heat treatment, green proteins solubility increased significantly, reaching 89.74% at pH 8, nearly 4.5 times higher than before treatment, showing low solubility only at the isoelectric point and excellent stability maintained in salt concentrations up to 1 M NaCl. A notable decrease in particle size was observed, from 1883 nm to 192 nm, leading to a more uniform particle size distribution. SDS-PAGE and FTIR deconvolution analyses indicated structural changes, possibly involving disulfide bond formation. The promising findings of this research, not only ensure complete biomass utilization, but also highlight the need for further investigation regarding the potential of green modified proteins, as the observed improvements open up numerous biotechnological applications. One potential direction is green protein-stabilized emulsions, supported by a significant increase in alpha-helix content associated with enhanced functional properties, which offers new possibilities for future research on stable and functional emulsions.</p>","PeriodicalId":9067,"journal":{"name":"Bioresources and Bioprocessing","volume":"12 1","pages":"102"},"PeriodicalIF":5.1,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12457265/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145124189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}