Bioresource Technology最新文献_第8页

Reliable biochemical methane potential testing: insights and recommendations from global interlaboratory study 可靠的生化甲烷潜力测试：来自全球实验室间研究的见解和建议

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-02-01 DOI: 10.1016/j.biortech.2026.134136

Sasha D. Hafner , Estelle Leca , Cecilia Sambusiti , Sergi Astals , Olivier Azam , Konrad Koch , Jing Liu , Henrik B. Møller , Mihaela Nistor , Mario Olaya-Rincon , Christine Peyrelasse , Alastair J. Ward , Bastien Zennaro , Florian Monlau

Thirty-one laboratories measured biochemical methane potential (BMP) of six substrates in a project aimed at improving BMP measurement quality. Laboratories used their established measurement procedures, with limited standardization. As in earlier studies, reproducibility was quantified, but here laboratories came from a wide geographic range, and the accuracy of BMP measurements was assessed by comparison with 6 experienced “reference” laboratories. Measurement reproducibility between the remaining “evaluation” laboratories was poor in the first test; relative reproducibility standard deviation was 21–33% for four substrates. Values improved to 9–19% in the second test following review of a detailed analysis of submitted data, troubleshooting meetings, and method modifications by some laboratories. Reference laboratories performed better, with reproducibility standard deviation of 5–12%. Repeatability within laboratories was also better for the reference group, with relative standard deviation of 2–4% vs. 3–7% for evaluation laboratories. Evaluation laboratory results showed a persistent negative bias of 4–16% compared to reference laboratories. Application of cellulose validation criteria improved reproducibility but not accuracy. Most of the extreme BMP values were associated with abnormal methane production curves, and a subjective visual evaluation proved valuable as a screening tool. The cause of measurement error was generally unknown, although data processing and volatile solids measurement errors caused some extreme results, and changes in inoculum source improved performance in two cases. There were no consistent differences among the diverse measurement methods and the sources of error probably vary among laboratories. Consistently accurate BMP measurement is possible for laboratories that follow established guidelines and develop experience.

在一个旨在提高BMP测量质量的项目中，31个实验室测量了6种底物的生化甲烷势（BMP）。实验室使用其既定的测量程序，标准化程度有限。在早期的研究中，重复性是量化的，但这里的实验室来自广泛的地理范围，BMP测量的准确性是通过与6个经验丰富的“参考”实验室进行比较来评估的。在第一次测试中，其余“评估”实验室之间的测量重复性较差；4种底物的相对重现性标准偏差为21-33%。在对提交数据的详细分析、故障排除会议和一些实验室的方法修改进行审查后，在第二次测试中，数值提高到9-19%。参考实验室表现较好，重复性标准偏差为5-12%。参照组实验室内的重复性也更好，相对标准偏差为2-4%，而评价实验室为3-7%。评价实验室结果显示与参比实验室相比持续存在5-16%的负偏倚。纤维素验证标准的应用提高了重现性，但没有提高准确性。大多数极端的BMP值与异常的甲烷产量曲线有关，主观的视觉评价被证明是有价值的筛选工具。测量误差的原因通常是未知的，尽管数据处理和挥发性固体测量误差会导致一些极端的结果，并且在两种情况下，接种源的改变提高了性能。不同的测量方法之间没有一致的差异，误差来源可能因实验室而异。对于遵循既定指南和开发经验的实验室来说，始终准确的BMP测量是可能的。

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引用次数: 0

Novel incomplete nitritation-regulated multi-pathway anammox process for advanced nitrogen removal from real municipal wastewater 新型不完全硝化调控的多途径厌氧氨氧化工艺用于实际城市污水的深度脱氮

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-02-06 DOI: 10.1016/j.biortech.2026.134167

Chengkun Kao, Qiong Zhang, Jinjin Liu, Wenyu Li, Yingtong Chen, Yongzhen Peng

Mainstream anammox continues to face fundamental operational bottlenecks, due to unstable nitrite supply and the limited competitive strength of autotrophic anammox bacteria in municipal wastewater. In this study, a novel strategy based on incomplete nitritation was established to initiate and regulate the multi-pathway anammox process. Short-term hydroxylamine induction (5 mg/L) rapidly established coexistence of NH₄⁺, NO₂⁻, and NO₃⁻, sustaining nitrite supply via nitritation (PN) and denitritation (PD). Advanced nitrogen removal (NRE = 93.3%) was achieved at extremely low C/N (2.5) during real municipal wastewater treatment. The activity and abundance of anammox bacteria increased markedly (6.2 → 16.7 mg NH₄⁺-N/gMLVSS/d; 5.6 × 10⁷→9.0 × 10⁸ gene copies/g VSS). Genomic and transcriptomic analyses revealed enriched Candidatus Brocadia (0.02%→0.18%) enhanced nitrogen removal while suppressing nitrite competition from nitrite oxidizing bacteria and denitrifiers, thereby strengthening both PN and PD pathways. Functional gene ratios demonstrated strong coordination between substrate supply pathways, with AOB/NOB increasing from 1.3 to 6.2 and (narG + napA)/(nirS + nirK) rising from 0.2 ± 0.1 to 1.0 ± 0.1. Importantly, incomplete rather than full nitrite oxidizing bacteria suppression was identified as a sustainable control point, ensuring robustness. Overall, multi-pathway anammox regulated by incomplete nitritation offers superior efficiency and stability compared to single-path systems, providing a promising strategy for mainstream anammox applications.

由于亚硝酸盐供应不稳定以及城市污水中自养厌氧氨氧化菌的竞争力有限，主流厌氧氨氧化仍然面临根本性的操作瓶颈。本研究建立了一种基于不完全硝化的新策略来启动和调节多途径厌氧氨氧化过程。短期羟胺诱导（5mg /L）迅速建立了NH4+、NO2−和NO3−的共存，通过硝化（PN）和反硝化（PD）维持亚硝酸盐的供应。在实际城市污水处理中，在极低的C/N（2.5）条件下实现了深度脱氮（NRE = 93.3%）。厌氧氨氧化菌活性和丰度显著增加（6.2→16.7 mg NH4+-N/gMLVSS/d; 5.6 × 107→9.0 × 108基因拷贝/g VSS）。基因组学和转录组学分析显示，富集的Candidatus Brocadia（0.02%→0.18%）增强了氮的去除，同时抑制了亚硝酸盐氧化菌和反硝化菌对亚硝酸盐的竞争，从而加强了PN和PD途径。AOB/NOB从1.3增加到6.2,(narG + napA)/(nirS + nirK)从0.2±0.1增加到1.0±0.1。重要的是，不完全而不是完全亚硝酸盐氧化细菌抑制被确定为可持续控制点，确保稳健性。总的来说，不完全硝化调控的多途径厌氧氨氧化比单途径系统具有更高的效率和稳定性，为主流厌氧氨氧化应用提供了一个有前景的策略。

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引用次数: 0

Potential of 2,3-butanediol, volatile fatty acid and methane production from medium and low-value forest industry side streams 中低值林产侧流生产2,3-丁二醇、挥发性脂肪酸和甲烷的潜力

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-24 DOI: 10.1016/j.biortech.2026.134089

Elina Tampio , Lucia Blasco , Milla Tynkkynen , Ilmari Laaksonen , Summaira Saghir , Petri Kilpeläinen , Reijo Lappalainen

Medium- and low-value side streams from the forest industry, such as sawdust and biosludge, are typically used for energy production, yet they offer potential for higher-value products via biological conversion. This study explores the combined production of 2,3-butanediol (BDO) and volatile fatty acids (VFAs) from birch sawdust and sludges from kraft mill pulping and wastewater treatment, alongside anaerobic digestion for methane. Integration of BDO and VFA production is evaluated by valorising BDO process residues. Pretreated birch sawdust yielded BDO 0.12 g/g of total solids (TS), and VFA up to 0.68 g/gTS, indicating economic feasibility. Combining both processes in a biorefinery could enhance revenues. Kraft mill sludges produced BDO up to 0.88 g/gTS, VFAs between 0.45–0.64 g/gTS, and methane 430 Nm³/t volatile solids (VS) without pretreatment. These findings demonstrate the potential of forest industry side streams for producing bio-based chemicals and methane, supporting integrated biorefineries for improved resource efficiency and value creation.

来自森林工业的中低价值侧流，如锯末和生物污泥，通常用于能源生产，但它们通过生物转化提供了生产高价值产品的潜力。本研究探讨了从桦木木屑和硫酸盐厂制浆和废水处理的污泥中联合生产2,3-丁二醇（BDO）和挥发性脂肪酸（VFAs），以及甲烷的厌氧消化。通过对BDO过程残留物进行估值，对BDO和VFA生产的整合进行评估。预处理后的桦木木屑总固体（TS） BDO为0.12 g/g， VFA为0.68 g/gTS，具有经济可行性。在生物精炼厂中结合这两种工艺可以增加收入。未经预处理的卡夫磨污泥BDO最高可达0.88 g/gTS， VFAs在0.45-0.64 g/gTS之间，甲烷挥发性固体（VS）为430 Nm3/t。这些发现表明，森林工业侧流在生产生物基化学品和甲烷方面具有潜力，支持综合生物精炼厂提高资源效率和创造价值。

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引用次数: 0

Multi-objective optimization of oxygen transfer and hydrodynamic shear in dual-impeller stirred tanks via integrated computational fluid dynamics and machine learning framework 基于综合计算流体力学和机器学习框架的双叶轮搅拌槽氧传递和流体动力剪切多目标优化

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-02-07 DOI: 10.1016/j.biortech.2026.134166

Qingfeng Gu , Shuoyan Ji , Yongqiang Liu , Yongbo Wang , Junxiong Yu , Zejian Wang , Yingping Zhuang , Chao Li

Optimizing the structure of stirred bioreactors is crucial to improve the efficiency of biomanufacturing. This study proposes an intelligent design framework integrating computational fluid dynamics (CFD), machine learning (ML), and the non-dominated sorting genetic algorithm II (NSGA-II) to co-optimize the oxygenation and shear performance of a dual-impeller stirred tank. Initially, a parameterized CFD model was employed to systematically investigate the impacts of impeller inclination angles, dimensions, and blade numbers on the volumetric oxygen transfer coefficient (

k_{L} a

) and shear strain rate. Subsequently, six ML algorithms were evaluated using a dataset of 837 CFD-generated samples, where the artificial neural network (ANN) demonstrated superior predictive accuracy (R² > 0.97) and was selected as the surrogate model. Utilizing the NSGA-II algorithm coupled with the ideal point decision-making method, an optimal asymmetric impeller configuration was identified from the Pareto-optimal frontier. The results reveal that the optimized design enhanced

k_{L} a

by 98.4% (reaching 312.5 h⁻¹) while maintaining a shear intensity comparable to the baseline. Finally, fermentation experiments with Blakeslea trispora confirmed that the optimized configuration led to a 30.9% increase in β-carotene yield (attaining 3193 mg/L). This work provides a robust and efficient methodology for the rational design of bioreactors and offers a new pathway for the intensification of shear-sensitive bioprocesses.

搅拌反应器结构的优化是提高生物制造效率的关键。本研究提出了一种集成计算流体力学（CFD）、机器学习（ML）和非主导排序遗传算法II （NSGA-II）的智能设计框架，以共同优化双叶轮搅拌槽的充氧和剪切性能。首先，采用参数化CFD模型，系统研究了叶轮倾角、尺寸和叶片数对体积氧传递系数（kLa）和剪切应变率的影响。随后，使用837个cfd生成样本的数据集对六种ML算法进行了评估，其中人工神经网络（ANN）表现出更高的预测精度（R2 > 0.97），并被选为代理模型。利用NSGA-II算法结合理想点决策方法，从pareto最优边界出发，确定了非对称叶轮的最优配置。结果表明，优化后的kLa提高了98.4%（达到312.5 h−1），同时保持了与基线相当的剪切强度。最后，对三孢黑霉进行发酵实验，结果表明，优化后的配置可使β-胡萝卜素产量提高30.9%，达到3193 mg/L。本研究为生物反应器的合理设计提供了可靠有效的方法，并为强化剪切敏感生物过程提供了新的途径。

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引用次数: 0

Optimizing synergistic microalgae cultivation and photocatalysis processes for antibiotic and nutrient removal in wastewater 优化微藻培养和光催化协同去除废水中抗生素和营养物的工艺

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-31 DOI: 10.1016/j.biortech.2026.134141

Lin-Lan Zhuang , Hongyang Liang , Weiyi Qian , Zhen Hu , Xiao-Xiong Wang , Jian Zhang

Coupling microalgae cultivation and photocatalysis processes exhibits potential for multi-pollutant removal from wastewater. However, the interaction between microalgae and photocatalysis has yet to be investigated. We revealed the indirect function of Chlorella Vulgaris to g-C₃N₄ could enhance up to 25 % sulfamerazine (SMZ) removal through the secretion of dissolved oxygen (DO), extracellular polymeric substances (EPS), and alkaline substances. Additionally, g-C₃N₄ degraded SMZ to reduce their toxicity to Chlorella Vulgaris growth. However, the shading effect of the Chlorella Vulgaris and the intrinsic toxicity of g-C₃N₄ could reduce the each other’s performance within a suspension system. Therefore, we designed an attachment-based photoreactor to avoid direct contact between them and maximize the mutual benefits of their secreted substances. Compared to the suspension system, this photoreactor significantly improved the removal of SMZ (5 mg/L), nitrogen (N) and phosphorous (P) by 57 %, 495 % and 112 %, respectively, with operation costs reduced by 66 %, revealing the superiority of this design.

微藻培养和光催化耦合过程具有去除废水中多种污染物的潜力。然而，微藻与光催化之间的相互作用尚未得到深入的研究。研究发现，小球藻对g-C3N4的间接作用可以通过分泌溶解氧（DO）、胞外聚合物质（EPS）和碱性物质，提高高达25%的磺胺嘧啶（SMZ）的去除率。此外，g-C3N4还能降解SMZ，降低其对小球藻生长的毒性。然而，小球藻的遮荫效应和g-C3N4的内在毒性会在悬浮体系中相互降低性能。因此，我们设计了一种基于附着的光反应器，以避免它们之间的直接接触，并最大限度地发挥它们的分泌物质的相互效益。与悬浮系统相比，该光反应器对SMZ （5 mg/L）、氮(N)和磷(P)的去除率分别提高了57%、495%和112%，运行成本降低了66%，显示了该设计的优越性。

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引用次数: 0

Integrated dynamic control and enzyme co-localization strategies enable high-efficiency stilbenoid biosynthesis 集成的动态控制和酶共定位策略实现了高效的二苯乙烯类生物合成

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-30 DOI: 10.1016/j.biortech.2026.134122

Guofu Zhao , Pengpai Li , Xinyao Ding , Jiugong Lv , Ying Yang , Lin Liu , Lihan Zi , Qiang Zhang , Liya Liang , Rongming Liu

Stilbenoids are plant-derived polyphenols with potent bioactivities, which makes them promising for pharmaceutical and nutraceutical use. Their production remains challenging due to their low natural abundance in plants and the inherent toxicity associated with chemical synthesis. Here, we developed an integrated strategy to engineer Escherichia coli for efficient biosynthesis of multiple stilbenoids. A p-coumaric acid-responsive CRISPRi system dynamically regulated malonyl-CoA allocation, balancing host metabolism and stilbenoid production. We systematically explored enzyme co-localization strategies and used structure-informed in silico analysis to support dual-enzyme fusion design. Orthogonal SpyTag/SpyCatcher and SnoopTag/SnoopCatcher systems enabled optimal intermediate channeling and significantly enhanced pathway flux. As a result, we achieved the highest reported titers of piceatannol (494.30 mg/L from glycerol, increased to 583.10 mg/L with l-tyrosine supplementation) and pterostilbene (563.89 mg/L from glycerol, reaching 1110.92 mg/L with l-tyrosine feeding). Techno-economic analysis indicated that combined optimization of metabolic regulation, enzyme co-localization, and process design contributes substantially to the overall feasibility of stilbenoid manufacturing. Altogether, this study demonstrates the potential of stilbenoid biosynthesis and provides a valuable paradigm for the efficient synthesis of other complex natural products.

二苯乙烯类化合物是植物源性多酚类物质，具有较强的生物活性，在医药和营养保健方面具有广阔的应用前景。由于它们在植物中的天然丰度较低，并且与化学合成相关的固有毒性，它们的生产仍然具有挑战性。在这里，我们开发了一种整合的策略来改造大肠杆菌，使其高效地合成多种苯乙烯类化合物。对香豆酸敏感的CRISPRi系统动态调节丙二酰辅酶A的分配，平衡宿主代谢和二苯乙烯类化合物的产生。我们系统地探索了酶共定位策略，并使用结构信息的硅分析来支持双酶融合设计。正交SpyTag/SpyCatcher和SnoopTag/SnoopCatcher系统实现了最佳的中间通道，并显著增强了通路通量。结果，我们获得了最高的报道滴度（甘油为494.30 mg/L，添加L -酪氨酸后增加到583.10 mg/L）和紫菜芪（甘油为563.89 mg/L，添加L -酪氨酸后达到1110.92 mg/L）。技术经济分析表明，代谢调控、酶共定位和工艺设计的组合优化大大提高了二苯乙烯类化合物生产的总体可行性。总之，本研究证明了二苯乙烯类生物合成的潜力，并为其他复杂天然产物的高效合成提供了有价值的范例。

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引用次数: 0

Competitive nitrogen/phosphorus scarcity triggers multi-pathway nitrogen removal and phosphorus-stress response in microalgae-bacterial symbiosis: poly-metabolic synergy enhancement mechanisms 竞争性氮磷稀缺触发微藻-细菌共生多途径脱氮和磷胁迫反应：多代谢协同增强机制

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-30 DOI: 10.1016/j.biortech.2026.134138

Lili Yang , Wenqian Wei , Xin Sun , Xiang Li , Xiaowen Wen , Binchen Mao , Ran Hao , Weihan Qiu , Huipeng Li

This study elucidates the metabolic remodeling and synergistic mechanisms of the microalgae-bacteria symbiosis (MABS) under competitive nitrogen and phosphorus stress. Compared with the control, MABS increased removal efficiencies of total nitrogen, NH₄⁺-N, NO₃⁻-N, and total phosphorus by 5.9, 5.1, 1.5, and 1.7 times, respectively, while enhancing microalgal triacylglycerol production by 17.5%. Microalgae preferentially assimilated NH₄⁺-N and dominated phosphorus uptake, whereas the bacterial community strengthened denitrification through functional specialization, carbon metabolism optimization, and electron transport chain (ETC) remodeling. Specifically, bacterial carbon metabolism shifted from the conventional tricarboxylic acid cycle toward a frdABCD-dependent branch that supplies reducing power, accompanied by the enrichment of electron carrier ubiquinone-10 and upregulation of ETC complexes III/IV. Symbiotic bacteria further promoted more efficient ATP synthesis in microalgae, synergizing with improved carbon fixation and lipid-directed carbon partitioning. These findings reveal the metabolic plasticity and cross-kingdom coordination that underpin high-rate nutrient removal and lipid accumulation in MABS.

本研究旨在阐明竞争氮磷胁迫下微藻-细菌共生（MABS）的代谢重塑及其协同机制。与对照相比，MABS对总氮、NH4+-N、NO3−-N和总磷的去除率分别提高了5.9倍、5.1倍、1.5倍和1.7倍，微藻的甘油三酯产量提高了17.5%。微藻优先吸收NH4+-N并主导磷的吸收，而细菌群落通过功能特化、碳代谢优化和电子传递链（ETC）重塑加强反硝化作用。具体来说，细菌的碳代谢从传统的三羧酸循环转变为提供还原能力的依赖于frdabcd的分支，伴随着电子载体泛素-10的富集和ETC复合物III/IV的上调。共生细菌进一步促进了微藻中更有效的ATP合成，与改进的碳固定和脂质定向碳分配协同作用。这些发现揭示了代谢可塑性和跨界协调是maabs中高速率营养去除和脂质积累的基础。

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引用次数: 0

Start-up of novel pilot-scale plant based on two-stage expanded granular sludge bed reactors for pig slurry treatment: Performance and microbial dynamics 基于两级膨胀颗粒污泥床反应器的新型猪浆处理中试装置的启动：性能和微生物动力学

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-22 DOI: 10.1016/j.biortech.2026.134075

Antonio J. Aragón-Barroso , Manuel J. Gallardo-Altamirano , Antonio Castellano-Hinojosa , Alejandro González-Martínez , Jesús González-López , Francisco Osorio

Pig manure management is an environmental challenge that can be improved through high-rate anaerobic digestion, enabling efficient biogas production and resource recovery. This study evaluated a pilot-scale two-stage expanded granular sludge bed (EGSB) system treating real pig slurry over a 150-day start-up. The reactors, operated in series with different working volumes (100 and 500 L), underwent progressive hydraulic retention times (HRT) reductions (R1: 3–1 d; R2: 12–7 d), increasing organic loading rates (OLR) (R1: 7.6–21.4 kg COD m⁻³ d⁻¹; R2: 1.0–2.5 kg COD m⁻³ d⁻¹). General chemical oxygen demand (COD) removal reached 68%, with system sensitivity to high solids, while biogas production and methane yield (MY) remained stable, reaching 264 L/d and 352 L CH₄/kg COD removed. Microbial analysis identified Clostridium sensu stricto_1, Methanosarcina, and Methanoculleus as key taxa supporting process stability. These results demonstrate the potential of two-stage EGSB systems for sustainable pig manure valorisation.

猪粪管理是一项环境挑战，可以通过高速率厌氧消化来改善，从而实现高效的沼气生产和资源回收。本研究评估了一个中试规模的两级膨胀颗粒污泥床（EGSB）系统，该系统在150天的启动过程中处理真实的猪浆。反应器以不同的工作容积（100和500 L）串联运行，经历了水力滞留时间（HRT）的逐步降低（R1: 3 - 1 d; R2: 12-7 d），提高了有机负载率（OLR）（R1: 7.6-21.4 kg COD m−3 d−1;R2: 1.0-2.5 kg COD m−3 d−1）。一般化学需氧量（COD）去除率达到68%，系统对高固体敏感，而沼气产量和甲烷产量（MY）保持稳定，分别达到264 L/d和352 L CH4/kg COD去除率。微生物分析表明，严格感梭菌（Clostridium sensu stricto_1）、产甲烷菌（Methanosarcina）和产甲烷菌（Methanoculleus）是支持工艺稳定性的关键类群。这些结果证明了两级EGSB系统在猪粪可持续增值方面的潜力。

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引用次数: 0

Engineering stress tolerance in Saccharomyces cerevisiae by overexpressing PIR3 and SPI1 for efficient ethanol production from high-concentration sugarcane molasses 通过过表达PIR3和SPI1，从高浓度甘蔗糖蜜中高效生产乙醇，来改造酿酒酵母的抗逆性

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-02-09 DOI: 10.1016/j.biortech.2026.134191

Wei-Yang Wang , Bei Liao , Ping Zheng , Ming-Yue Huang , Yu-Tuo Wei , Fu-Xing Niu

The coexistence of potassium and calcium ions has been identified as a major factor limiting high-yield ethanol fermentation by Saccharomyces cerevisiae in high-concentration sugarcane molasses. To identify key genes conferring tolerance to this stress, we employed an integrated strategy combining multi-omics analysis, CRISPR-mediated gene activation/repression, and targeted overexpression. This approach pinpointed four critical genes: PIR3, SPI1, AQR1, and GUT2. Functional analysis showed that while AQR1 and GUT2 enhance ethanol biosynthesis primarily by redirecting metabolic flux, PIR3 and SPI1 are crucial for maintaining cellular integrity and viability under stress. Overexpression of PIR3 and SPI1 in a wild-type strain increased ethanol production by 24.6%, achieving a final titer of 113.3 g/L in a 5-L fermenter, a performance comparable to robust industrial strains. Furthermore, this engineering strategy boosted the synthesis of other valuable compounds, exemplified by a 12.5% increase in cinnamic acid production. Our work thus identifies precise genetic targets for engineering stress-tolerant yeast and establishes a foundation for efficient bioconversion of high-concentration sugarcane molasses.

钾离子和钙离子的共存是制约酿酒酵母在高浓度甘蔗糖蜜中高产乙醇发酵的主要因素。为了确定对这种胁迫具有耐受性的关键基因，我们采用了一种综合策略，结合多组学分析、crispr介导的基因激活/抑制和靶向过表达。这种方法确定了四个关键基因：PIR3、SPI1、AQR1和GUT2。功能分析表明，虽然AQR1和GUT2主要通过重定向代谢通量来促进乙醇生物合成，但PIR3和SPI1在逆境下维持细胞完整性和活力至关重要。PIR3和SPI1在野生型菌株中过表达可使乙醇产量提高24.6%，在5-L发酵罐中最终滴度达到113.3 g/L，性能与强劲的工业菌株相当。此外，这种工程策略促进了其他有价值化合物的合成，例如肉桂酸的产量增加了12.5%。因此，我们的工作确定了工程抗逆性酵母的精确遗传靶点，并为高浓度甘蔗糖蜜的高效生物转化奠定了基础。

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引用次数: 0

Occurrence, persistence and vertical distribution of high-risk antibiotic resistance genes in biogas slurry-amended soils across China 中国沼液改良土壤中高危抗生素耐药基因的发生、持续及垂直分布

IF 9 1区环境科学与生态学 Q1 AGRICULTURAL ENGINEERING

Bioresource Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-24 DOI: 10.1016/j.biortech.2026.134088

Liting Zhang , Hao Li , Jiwei Li , Yi Lu , Yanjun Chai , Zichuan Li , Lifeng Ping , Ming Hung Wong , Shengdao Shan

Various multidrug-resistant bacteria carrying high-risk antibiotic resistance genes (ARGs) occurred in agricultural soils amended with biogas slurry, threatening human health by exhibiting resistance to clinically significant antibiotics. However, it remains unclear whether high-risk ARGs occur and persist widely in biogas slurry-amended soils, and few studies have further focused on their vertical distribution. Thus, the present study investigated the occurrence, persistence and vertical distribution of high-risk ARGs in biogas slurry-amended soils from Hebei, Shaanxi, Gansu and Zhejiang districts across China. The obtained results indicate that high-risk ARGs and their associated MGEs were widely persisted and enriched in agricultural soils amended with biogas slurry, especially intI1, tetracycline, aminoglycoside and phenicol ARGs. The potentially vertical distribution of enriched high-risk ARGs depended on their subtypes and district background, and the tet(L), fexA, erm(A), tet(B), dfrA12, tet(A), aadA1, aac(6′)-Ib3 and floR were further enriched with increasing soil depth. ARG-associated bacteria dominated their persistence and vertical distribution in biogas slurry-amended soils. Furthermore, biogas slurry application conditions and regional climate both play important roles in shaping ARGs-associated bacteria community in the soil environment, and cropland characteristics, along with biogas slurry application conditions/regional climate, evidently influenced the vertical distribution of ARGs-associated bacteria. The obtained results help improving the treatments and disposal of animal manure, to limit the inputs and subsequent diffusion of high-risk ARGs from livestock farms into the agricultural soil–water environment.

在沼液改良的农业土壤中，多种携带高风险抗生素耐药基因（ARGs）的多重耐药细菌出现，对临床重要抗生素产生耐药性，威胁人类健康。然而，目前尚不清楚高风险ARGs是否会在沼气浆改性土壤中广泛发生和持续存在，并且很少有研究进一步关注其垂直分布。因此，本研究对中国河北、陕西、甘肃和浙江4个地区沼气浆改良土壤中高危ARGs的发生、持续和垂直分布进行了研究。结果表明，在沼液改良的农业土壤中，高风险ARGs及其相关的MGEs广泛存在并富集，特别是ti1、四环素、氨基糖苷类和酚类ARGs。高风险ARGs的潜在垂直富集分布取决于其亚型和地区背景，tet(L)、fexA、erm(A)、tet(B)、dfrA12、tet(A)、aadA1、aac（6′）-Ib3和floR随着土壤深度的增加而进一步富集。在沼液改良土壤中，arg相关菌的持久性和垂直分布占主导地位。此外，沼液施用条件和区域气候对土壤环境中args相关细菌群落的形成都有重要影响，农田特征以及沼液施用条件/区域气候对args相关细菌的垂直分布有明显影响。所得结果有助于改善动物粪便的处理和处置，以限制高风险ARGs从畜牧场进入农业土壤-水环境及其随后的扩散。

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