Hao Zhang , Hairui Tong , Qiang Yin , Yibin Qiu , Hong Xu , Sha Li
{"title":"Efficient production of ectoine from Jerusalem artichoke using engineered Escherichia coli","authors":"Hao Zhang , Hairui Tong , Qiang Yin , Yibin Qiu , Hong Xu , Sha Li","doi":"10.1016/j.biortech.2025.132589","DOIUrl":"10.1016/j.biortech.2025.132589","url":null,"abstract":"<div><div>In this study, a recombinant <em>Escherichia coli</em> strain was constructed to produce ectoine from Jerusalem artichoke through modular pathway engineering. First, a promoter-optimized ectoine synthesis module was integrated into the chromosome using multiple copies. Then, the introduction and expression of inulin hydrolase was optimized because inulin cannot be directly utilized. Subsequently, Fructose transport and phosphorylation, glycolysis, and oxaloacetate supply module were enhanced separately and in combination to improve ectoine production and substrate utilization. The strain ETC16 (co-expression of <em>gapA</em>, <em>ppc</em>, and <em>fruK</em>, Δ<em>iclR</em>) produced 6.51 g/L ectoine with 0.13 g/g inulin. Furthermore, the raw inulin extract and monosodium glutamate (MSG) residue were optimized for ectoine production. Finally, 35.60 g/L of ectoine with a yield of 0.36 g/g inulin was achieved in a 7.5 L fermenter. This study revealed a potential method of non-food fermentation to produce high-value products.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"431 ","pages":"Article 132589"},"PeriodicalIF":9.7,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143900120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Wanying Li, Rui Huo, Yiling Di, Chun Liu, Shilei Zhou
{"title":"Efficient nitrogen removal by the aerobic denitrifying bacterium Pseudomonas stutzeri RAS-L11 under triple stresses of high alkalinity, high salinity, and tetracycline: From performance to mechanism","authors":"Wanying Li, Rui Huo, Yiling Di, Chun Liu, Shilei Zhou","doi":"10.1016/j.biortech.2025.132590","DOIUrl":"10.1016/j.biortech.2025.132590","url":null,"abstract":"<div><div>Efficient aerobic denitrification bacteria are rarely reported under triple stresses of high alkalinity, high salinity, and tetracycline. Here, strain <em>Pseudomonas stutzer</em>i RAS-<em>L</em>11 was isolated, under the optimal reaction conditions of C/N = 6, sodium acetate as carbon source, and pH 7.0–11.0. Moreover, RAS-<em>L</em>11 showed perfect nitrogen removal performance under dual and triple stresses. Specifically, the mean removal efficiency of total dissolved nitrogen for different medium (nitrate, nitrite, ammonia, nitrate and ammonia, and nitrite and ammonia) reached 92.35 %, 66.85 %, 71.33 %, 89.42 %, and 68.76 % under triple stresses. Nitrogen balance results indicated that biomass nitrogen accounted for a small percentage (4.48 % to 20.79 %). Furthermore, the nitrogen metabolism pathways and tetracycline, salinity, and alkaline tolerance-associated genes were also confirmed. Strain RAS-<em>L</em>11 achieved 42.67–70.72 % NO<sub>3</sub><sup>−</sup>-N and 83.72–88.53 % NH<sub>4</sub><sup>+</sup>-N removal efficiencies in both sterilized and actual systems treating pharmaceutical wastewater. Our characterization of the RAS-<em>L</em>11 provides a reference for nitrogen removal of pharmaceutical wastewater.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132590"},"PeriodicalIF":9.7,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143882612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mi Tang , Zhenhui Pan , Minghui Jin , Hengwei Zhang , Xuewei Pan , Zhiming Rao
{"title":"Systems metabolic engineering of Escherichia coli for the high-level production of deoxyviolacein, a natural colorant","authors":"Mi Tang , Zhenhui Pan , Minghui Jin , Hengwei Zhang , Xuewei Pan , Zhiming Rao","doi":"10.1016/j.biortech.2025.132584","DOIUrl":"10.1016/j.biortech.2025.132584","url":null,"abstract":"<div><div>Deoxyviolacein is a natural colorant with various biological properties, widely applied in cosmetic and pharmaceutical fields. However, current methods of deoxyviolacein production by natural producers may cause highly lethal infections in humans, limiting the sustainable production of deoxyviolacein. Here, an <span>l</span>-tryptophan-producing <em>Escherichia coli</em> strain was engineered for efficient deoxyviolacein production. First, the deoxyviolacein synthesis pathway was introduced and optimized to construct a base strain. Second, multi-modular engineering was conducted for further optimization, including engineering of the glucose uptake system and central metabolism and enhancement of precursor supply. To coordinate metabolic flux distribution, the optimal expression of <em>aroG</em><sup>Q151F</sup>, <em>ppsA</em> and <em>tktA</em> was tuned by generating libraries of tunable intergenic regions coupled with a novel <span>l</span>-tryptophan biosensor. Finally, the best-performing strain successfully accumulated 12.18 g/L of deoxyviolacein from glucose, showing a competitive deoxyviolacein titer reported to date and providing a paradigm for the production of value-added aromatic compounds in <em>E. coli</em>.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"431 ","pages":"Article 132584"},"PeriodicalIF":9.7,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143894429","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yanming Qiao , Ehsan Kargaran , Hao Ji , Meysam Madadi , Saeed Rafieyan , Dan Liu
{"title":"Data-driven insights for enhanced cellulose conversion to 5-hydroxymethylfurfural using machine learning","authors":"Yanming Qiao , Ehsan Kargaran , Hao Ji , Meysam Madadi , Saeed Rafieyan , Dan Liu","doi":"10.1016/j.biortech.2025.132582","DOIUrl":"10.1016/j.biortech.2025.132582","url":null,"abstract":"<div><div>Converting cellulose into 5-Hydroxymethylfurfural (HMF) provides a promising strategy for creating bio-based chemicals, offering sustainable alternatives to petroleum-based materials in polymers, biofuels, and pharmaceuticals. However, the efficient production of HMF from cellulose is challenged by the complex interplay of numerous operational variables. This study develops a machine learning (ML) model to optimize HMF production and conducts a feature importance analysis to identify the key factors affecting HMF yield. Additionally, a Bayesian optimization is employed for multi-objective optimization aimed at maximizing HMF yield. A comprehensive dataset, sourced from existing literature, was subjected to statistical analysis to elucidate the influence of each factor on HMF production. Among the eight models evaluated, the CatBoost Regressor emerged as the most effective, delivering robust predictive performance with R<sup>2</sup> of 0.76 during testing and exhibiting low RMSE (4.72) and MAE (5.2) values. Feature importance analysis revealed that operational conditions, particularly time and temperature, were the most significant, accounting for 41.0% of the variability, followed by catalyst properties at 33.0% and solvent properties at 26.0%. The ML-based optimization achieved an HMF yield of 48.1%, with relative errors of −1% and 2.5% in the first (47.6%) and second (49.3%) runs of experimental validation, respectively. This research showcases ML’s ability to address challenges in cellulose-to-HMF conversion, offering insights for optimizing production and advancing sustainable manufacturing.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132582"},"PeriodicalIF":9.7,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143882713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Investigation of mannitol as a potential substrate for production of 2′-fucosyllactose in Yarrowia lipolytica","authors":"Yan Zhang, Haiyan Liu, Mengmeng Liu, Qingsheng Qi","doi":"10.1016/j.biortech.2025.132583","DOIUrl":"10.1016/j.biortech.2025.132583","url":null,"abstract":"<div><div>Mannitol, boasting abundant availability in marine biomass resource, represents a promising feedstock for biomanufacturing. Herein, <em>Yarrowia lipolytica</em> was found to synthesize 2′-fucosyllactose (2′-FL) more efficiently when using mannitol and lactose as substrate, compared to glucose and lactose. Notably, the strain maintained robust growth and 2′-FL production even when cultivated in brown algae processing wastewater. Transcriptome analysis of <em>Y. lipolytica</em> grown on mannitol revealed that two endogenous hexose transporters (Yht1 and Yht3) potentially modulated mannitol uptake. By optimizing the mannitol transporter and the GDP-<span>d</span>-mannose synthesis pathway, a 2′-FL producing strain was constructed with a titer of 9 g/L. Fed-batch fermentation in an inorganic salt medium supplemented with mannitol and lactose elevated the 2′-FL titer to 27.6 g/L. Collectively, this study demonstrates the efficacy of mannitol for 2′-FL biosynthesis and advances the sustainable utilization of algal resources.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132583"},"PeriodicalIF":9.7,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143877265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Optimization of synergistic microwave and zero–valent iron co–pretreatment for anaerobic digestion of waste activated sludge","authors":"Seung–han Ha , Seung Gu Shin , Johng–Hwa Ahn","doi":"10.1016/j.biortech.2025.132568","DOIUrl":"10.1016/j.biortech.2025.132568","url":null,"abstract":"<div><div>This study optimized co<strong>-</strong>pretreatment of microwave temperature (<em>T</em><sub>MW</sub>) and zero-valent iron dosage ([ZVI]) to enhance anaerobic digestion (AD) of waste activated sludge (WAS). WAS was pretreated at <em>T</em><sub>MW</sub> = 100, 150, or 200 °C and [ZVI] = 1, 3, or 5 g/L using a central composite design. Optimal co-pretreatment (<em>T</em><sub>MW</sub> = 168 °C and [ZVI] = 5 g/L) reduced the ratio of volatile solids (VS) to total solids by 21.5 %, increased the solubilization ratio seven–fold, removed 53.5 % of phosphate compared to WAS partly because of lignin fragmentation. Biochemical methane potential identified optimal conditions (<em>T</em><sub>MW</sub> = 164 °C and [ZVI] = 4.8 g/L), enhancing VS removal by 70.9 %, methane yield by 60 %, and reducing hydrogen sulfide by 82.4 % compared to Control. Kinetic analysis indicated 61 – 108 % increase in maximum methane production rate. Microbial analysis revealed increased acetoclastic methanogens and decreased hydrogenotrophic methanogens. Thus, microwave–ZVI co-pretreatment enhanced WAS biodegradability and AD efficiency.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132568"},"PeriodicalIF":9.7,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhiyuan Du , Sai Bai , Jin Qian , Peng Zhan , Fengping Hu , Xiaoming Peng
{"title":"Iron-carbon enhanced constructed wetland microbial fuel cells for tetracycline wastewater treatment: Efficacy, power generation, and the role of iron-carbon","authors":"Zhiyuan Du , Sai Bai , Jin Qian , Peng Zhan , Fengping Hu , Xiaoming Peng","doi":"10.1016/j.biortech.2025.132578","DOIUrl":"10.1016/j.biortech.2025.132578","url":null,"abstract":"<div><div>Tetracycline (TC) antibiotics wastewater is a serious threat to human health and environment. In this study, four groups of laboratory-scale constructed wetlands (CWs) with different configurations were constructed to evaluate the removal efficiency of iron-carbon (Ic) coupled constructed wetland microbial fuel cells (CW-MFC) system for different pollutants removal and bioelectricity production. The results showed that the addition of Ic significantly promoted the removal of contaminants. The maximum removal rates of COD, TN, NH<sub>4</sub><sup>+</sup>-N, and TP were 86.13 %, 81.60 %, 79.07 %, and 97.35 %, respectively. In particular, the removal rates of TC reached 100 %. 3D-EEM analysis further confirmed the role of Ic in promoting organic degradation. The Ic-CW-MFC system also showed superiority in power generation performance with peak power density of 7.90 mW/m<sup>2</sup> (internal resistance is 10 Ω), 88.07 % higher than the traditional CW-MFC, while the internal resistance was 68.21 % lower. Therefore, when Ic is used as the substrate of CW-MFC system, its decontamination and electricity generation performance is the best. Analysis of RDA was used to elucidate the relationship of four CWs, dominant strains and environmental factors (pH, ORP and DO). The performance of traditional CWs decreased significantly after TC addition (5–20 mg/L), but Ic-CW-MFC could effectively alleviate the inhibition effect caused by high-concentration TC wastewater. The working mechanism of Ic-CW-MFC in TC wastewater was further analyzed through typical cycle experiment and characterization. The results showed that Ic-CW-MFC is an efficient and economical wastewater treatment technology, which has great potential application value in the treatment of wastewater containing TC.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132578"},"PeriodicalIF":9.7,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143868932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Luigi Marra , Elena Aurino , Francesca Raganati , Antonino Pollio , Antonio Marzocchella
{"title":"Selection and characterization of acid-tolerant Chlorophyta division microalgae for potential metabolites production in a bubble column bioreactor: A ready-to-industrial scale screening","authors":"Luigi Marra , Elena Aurino , Francesca Raganati , Antonino Pollio , Antonio Marzocchella","doi":"10.1016/j.biortech.2025.132579","DOIUrl":"10.1016/j.biortech.2025.132579","url":null,"abstract":"<div><div>Microalgae are promising candidates for sustainable bioprocesses owing to their ability to capture/utilize CO<sub>2</sub> and produce valuable metabolites. However, strain-to-strain variability poses a challenge to industrial scalability. This paper reports a study on the screening of 28 Chlorophyta microalgal strains from six species cultivated under standardized semi-continuous conditions in a bubble column bioreactor. Growth kinetics, nitrogen uptake, and metabolite accumulation were analyzed to identify strains characterized by promising biomass/metabolite productivities.</div><div>The results indicated two distinct kinetic behaviors: a large fraction of the investigated strains was characterized by a common biomass/nitrate uptake behavior, and a subset of the strain pool was characterized by atypical nitrogen assimilation patterns. Strains characterized by high biomass growth rates are characterized by high lipid productivity. Strains with intermediate biomass growth rates are characterized by high carbohydrate accumulation. <em>Chlorella vulgaris</em> (ACUF 058) and <em>Chloroidium saccharophilum</em> (ACUF 050) emerged as strong candidates for carbohydrate and protein production, respectively.</div><div>This study proposed a systematic procedure for strain selection, bridging laboratory screening, and industrial feasibility. These results emphasize the role of tailored cultivation strategies in optimizing biomass yield and metabolite productivity. Further research integrating genomic and proteomic insights will enhance strain-specific process optimization, facilitating the large-scale deployment of microalgal bioprocesses.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132579"},"PeriodicalIF":9.7,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143868934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lan Xu , Hui-Min Wang , Feng-Yi Li , Wen-Hui Zhao , Dan Yue , Zhen-Ming Chi , Guang-Lei Liu
{"title":"Enhanced expression of glycolysis regulators KmGcr1p and KmGcr2p in Kluyveromyces marxianus: An efficient strategy for high-temperature ethanol production from inulin","authors":"Lan Xu , Hui-Min Wang , Feng-Yi Li , Wen-Hui Zhao , Dan Yue , Zhen-Ming Chi , Guang-Lei Liu","doi":"10.1016/j.biortech.2025.132559","DOIUrl":"10.1016/j.biortech.2025.132559","url":null,"abstract":"<div><div>Bioethanol production from non-food biomass has emerged as a sustainable alternative to satisfy future energy demands. <em>Kluyveromyces marxianus</em> stands out as a potential consolidated bioprocessing (CBP) strain for high-temperature ethanol production from inulin. Nevertheless, enhancing its fermentation efficiency remains crucial. Here, we characterized its glycolysis regulator KmGcr2p and demonstrated that KmGcr2p boosts the expression of target genes controlled by another glycolysis regulator, KmGcr1p, through their interaction. Furthermore, we engineered the strain Ogcr1 + 2 through co-overexpressing <em>KmGCR1</em> and <em>KmGCR2</em>, leading to a 55.8 % increase in inulinase activity, improved growth at 37 °C to 45 °C and under 10 % ethanol, and enhanced ethanol production at 42 °C by 36.0 % from inulin and 39.5 % from Jerusalem artichoke tuber flour. Our results highlight that co-overexpression of glycolysis regulators KmGcr1p and KmGcr2p can simultaneously enhance inulin saccharification, thermotolerance, ethanol tolerance, and ethanol production, presenting a promising and feasible strategy for high-temperature CBP ethanol production from inulin and inulin-rich feedstocks.</div></div>","PeriodicalId":258,"journal":{"name":"Bioresource Technology","volume":"430 ","pages":"Article 132559"},"PeriodicalIF":9.7,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143868935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}