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Mutagenesis and fluorescence-activated cell sorting of oleaginous Saccharomyces cerevisiae and the multi-omics analysis of its high lipid accumulation mechanisms. 油酸酵母菌的突变和荧光激活细胞分选及其高脂积累机制的多组学分析。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-07-02 DOI: 10.1016/j.biortech.2024.131062
Xiaotong Ji, Lin Chen, Guanpin Yang, Chunlei Tang, Wenjun Zhou, Tianzhong Liu, Xuefeng Lu

Acquiring lipid-producing strains of Saccharomyces cerevisiae is necessary for producing high-value palmitoleic acid. This study sought to generate oleaginous S. cerevisiae mutants through a combination of zeocin mutagenesis and fluorescence-activated cell sorting, and then to identify key mutations responsible for enhanced lipid accumulation by multi-omics sequencing. Following three consecutive rounds of mutagenesis and sorting, a mutant, MU310, with the lipid content of 44%, was successfully obtained. Transcriptome and targeted metabolome analyses revealed that a coordinated response involving fatty acid precursor biosynthesis, nitrogen metabolism, pentose phosphate pathway, ethanol conversion, amino acid metabolism and fatty acid β-oxidation was crucial for promoting lipid accumulation. The carbon fluxes of acetyl-CoA and NADPH in lipid biosynthesis were boosted in these pathways. Certain transcriptional regulators may also play significant roles in modulating lipid biosynthesis. Results of this study provide high-quality resource for palmitoleic acid production and deepen the understanding of lipid synthesis in yeast.

获得产脂酵母菌株是生产高价值棕榈油酸的必要条件。本研究试图通过玉米素诱变和荧光激活细胞分选相结合的方法产生产油酿酒酵母突变体,然后通过多组学测序鉴定导致脂质积累增强的关键突变。经过连续三轮诱变和分选,成功获得了脂质含量为 44% 的突变体 MU310。转录组和靶向代谢组分析表明,脂肪酸前体生物合成、氮代谢、磷酸戊糖途径、乙醇转化、氨基酸代谢和脂肪酸β-氧化的协调反应是促进脂质积累的关键。在这些途径中,脂质生物合成过程中乙酰-CoA 和 NADPH 的碳通量增加。某些转录调节因子也可能在调节脂质生物合成中发挥重要作用。这项研究的结果为棕榈油酸的生产提供了优质资源,并加深了人们对酵母脂质合成的认识。
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引用次数: 0
Sustainable production of a biotechnologically relevant β-galactosidase in Escherichia coli cells using crude glycerol and cheese whey permeate. 利用粗甘油和奶酪乳清渗透物在大肠杆菌细胞中可持续地生产与生物技术相关的 β-半乳糖苷酶。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-07-02 DOI: 10.1016/j.biortech.2024.131063
Bianchi Greta, Pessina Alex, Ami Diletta, Signorelli Samuele, de Divitiis Marcella, Natalello Antonino, Lotti Marina, Brambilla Luca, Brocca Stefania, Mangiagalli Marco

Responsible use of natural resources and waste reduction are key concepts in bioeconomy. This study demonstrates that agro-food derived-biomasses from the Italian food industry, such as crude glycerol and cheese whey permeate (CWP), can be combined in a high-density fed-batch culture to produce a recombinant β-galactosidase from Marinomonas sp. Ef1 (M-βGal). In a small-scale process (1.5 L) using 250 mL of crude glycerol and 300 mL of lactose-rich CWP, approximately 2000 kU of recombinant M-βGal were successfully produced along with 30 g of galactose accumulated in the culture medium. The purified M-βGal exhibited high hydrolysis efficiency in lactose-rich matrices, with hydrolysis yields of 82 % in skimmed milk at 4 °C and 94 % in CWP at 50 °C, highlighting its biotechnological potential. This approach demonstrates the effective use of crude glycerol and CWP in sustainable and cost-effective high-density E. coli cultures, potentially applicable to recombinant production of various proteins.

负责任地利用自然资源和减少废物是生物经济的关键概念。本研究表明,来自意大利食品工业的农产食品衍生生物质,如粗甘油和奶酪乳清渗透物(CWP),可在高密度喂料批次培养中结合使用,以生产来自马林单胞菌 Ef1 的重组β-半乳糖苷酶(M-βGal)。在使用 250 mL 粗甘油和 300 mL 富含乳糖的 CWP 的小规模工艺(1.5 L)中,成功生产了约 2000 kU 重组 M-βGal,以及培养基中积累的 30 g 半乳糖。纯化的 M-βGal 在富含乳糖的基质中表现出很高的水解效率,在 4 °C 的脱脂奶中的水解率为 82%,在 50 °C 的 CWP 中的水解率为 94%,突显了其生物技术潜力。这种方法证明了在可持续和具有成本效益的高密度大肠杆菌培养物中有效利用粗甘油和 CWP,可用于重组生产各种蛋白质。
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引用次数: 0
Modelling anaerobic sulfide removal by sulfide shuttling bacteria. 利用硫化物穿梭细菌建立厌氧硫化物去除模型。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-07-02 DOI: 10.1016/j.biortech.2024.131064
Joris Bergman, Annemerel R Mol, Annemiek Ter Heijne, Karel J Keesman, Rikke Linssen

Sulfide oxidizing bacteria are used in industrial biodesulfurization processes to convert sulfide to sulfur. These bacteria can spatially separate sulfide removal from terminal electron transfer, thereby acting as sulfide shuttles. The mechanisms underlying sulfide shuttling are not yet clear. In this work, newly obtained sulfide removal data were used to develop a new model for anaerobic sulfide removal and this model was shown to be an improvement over two previously published models. The new model describes a fast chemical step and a consecutive slow enzymatic step. The improved model includes the effect of pH, with higher total sulfide removal at increasing pH, as well as partial sulfide removal at higher sulfide concentrations. The two-stage model is supported by recent developments in anaerobic sulfide removal research and contributes to a better understanding of the underlying mechanisms. The model is a step toward accurately modelling anaerobic sulfide removal in industrial systems.

硫化物氧化细菌用于工业生物脱硫过程,将硫化物转化为硫磺。这些细菌可以在空间上将硫化物脱除与终端电子转移分开,从而充当硫化物穿梭器。硫化物穿梭的内在机制尚不清楚。在这项工作中,新获得的硫化物去除数据被用于开发厌氧硫化物去除的新模型,该模型比之前发表的两个模型有了改进。新模型描述了一个快速化学步骤和一个连续的慢速酶步骤。改进后的模型包括 pH 值的影响,pH 值升高时硫化物的总去除率提高,硫化物浓度升高时硫化物的部分去除率也提高。两阶段模型得到了厌氧硫化物去除研究最新进展的支持,有助于更好地理解其基本机制。该模型朝着准确模拟工业系统中厌氧硫化物去除的方向迈出了一步。
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引用次数: 0
Engineering Bacillus licheniformis as industrial chassis for efficient bioproduction from starch 将地衣芽孢杆菌作为高效淀粉生物生产的工业底盘
IF 11.4 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-07-01 DOI: 10.1016/j.biortech.2024.131061
Jiang Zhu, Min Liu, Jianling Kang, Shiyi Wang, Ziyan Zha, Yangyang Zhan, Zhi Wang, Junhui Li, Dongbo Cai, Shouwen Chen
Starch is an attractive feedstock in biorefinery processes, while the low natural conversion rate of most microorganisms limits its applications. Herein, starch metabolic pathway was systematically investigated using DW2 as the host organism. Initially, the effects of overexpressing amylolytic enzymes on starch hydrolysis were evaluated. Subsequently, the transmembrane transport system and intracellular degradation module were modified to accelerate the uptake of hydrolysates and their further conversion to glucose-6-phosphate. The DW2-derived strains exhibited robust growth in starch medium, and productivity of bacitracin and subtilisin were improved by 38.5% and 32.6%, with an 32.3% and 22.9% increase of starch conversion rate, respectively. Lastly, the employment of engineering strategies enabled another WX-02 to produce poly-γ-glutamic acid from starch with a 2.1-fold increase of starch conversion rate. This study not only provided excellent chassis for sustainable bioproduction from starch, but shed light on researches of substrate utilization.
淀粉是生物精炼工艺中一种极具吸引力的原料,但大多数微生物的自然转化率较低,限制了其应用。本文以 DW2 为宿主生物,对淀粉代谢途径进行了系统研究。首先,评估了过表达淀粉分解酶对淀粉水解的影响。随后,对跨膜运输系统和细胞内降解模块进行了改造,以加速水解物的吸收并进一步转化为葡萄糖-6-磷酸。DW2 衍生菌株在淀粉培养基中生长旺盛,杆菌肽和枯草菌素的生产率分别提高了 38.5% 和 32.6%,淀粉转化率分别提高了 32.3% 和 22.9%。最后,利用工程策略,另一个 WX-02 从淀粉中生产聚-γ-谷氨酸,淀粉转化率提高了 2.1 倍。这项研究不仅为淀粉的可持续生物生产提供了很好的底盘,而且为底物利用的研究提供了启示。
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引用次数: 0
Selective production of high-value fuel via catalytic upgrading of bio-oil over nitrogen-doped carbon-alumina hybrid supported cobalt catalysts. 在掺氮碳-氧化铝杂化支撑钴催化剂上催化生物油升级,选择性生产高价值燃料。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-06-29 DOI: 10.1016/j.biortech.2024.131059
Qin Peng, Xia Jiang, Guangmei Cao, Tianqiao Xie, Ziheng Jin, Lingling Xie, Fengli Gan, Shenggui Ma, Mingming Peng

Bio-oil derived from biomass fast pyrolysis can be upgraded to gasoline and diesel alternatives by catalytic hydrodeoxygenation (HDO). Here, the novel nitrogen-doped carbon-alumina hybrid supported cobalt (Co/NCAn, n = 1, 2.5, 5) catalyst is established by a coagulation bath technique. The optimized Co/NCA2.5 catalyst presented 100 % conversion of guaiacol, high selectivity to cyclohexane (93.6 %), and extremely high deoxygenation degree (97.3 %), respectively. Therein, the formation of cyclohexanol was facilitated by stronger binding energy and greater charge transfer between Co and NC which was unraveled by density functional theory calculations. In addition, the appropriate amount of Lewis acid sites enhanced the cleavage of the C-O bond in cyclohexanol, finally resulting in a remarkable selectivity for cyclohexane. Finally, the Co/NCA2.5 catalyst also exhibited excellent selectivity (93.1 %) for high heating value hydrocarbon fuel in crude bio-oil HDO. This work provides a theoretical basis on N dopants collaborating alumina hybrid catalysts for efficient HDO reaction.

生物质快速热解产生的生物油可以通过催化加氢脱氧(HDO)升级为汽油和柴油替代品。在此,通过混凝浴技术建立了新型氮掺杂碳氧化铝杂化支撑钴(Co/NCAn,n = 1、2.5、5)催化剂。优化后的 Co/NCA2.5 催化剂的愈创木酚转化率为 100%,对环己烷的选择性高(93.6%),脱氧度极高(97.3%)。其中,Co 和 NC 之间更强的结合能和更大的电荷转移促进了环己醇的形成,密度泛函理论计算揭示了这一点。此外,适量的路易斯酸位点增强了环己醇中 C-O 键的裂解,最终导致了对环己烷的显著选择性。最后,Co/NCA2.5 催化剂对粗生物油 HDO 中的高热值碳氢化合物燃料也表现出卓越的选择性(93.1%)。这项研究为掺杂 N 的氧化铝杂化催化剂用于高效 HDO 反应提供了理论依据。
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引用次数: 0
Removal of artificial sweeteners in wastewater treatment plants and their degradation during sewage sludge composting with micro- and nano-sized kaolin 利用微纳米级高岭土去除污水处理厂中的人工甜味剂并在污水污泥堆肥过程中降解人工甜味剂。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-06-29 DOI: 10.1016/j.biortech.2024.131060
Jishao Jiang , Rui Hou , Huilin Cui , Zhuyu Tang , Yousif Abdelrahman Yousif Abdellah , Caspar C.C. Chater , Ke Cheng , Fuqiang Yu , Dong Liu

This study surveyed the fates of artificial sweeteners in influent, effluent, and sewage sludge (SS) in wastewater treatment plant, and investigated the effects of Micro-Kaolin (Micro-KL) and Nano-Kaolin (Nano-KL) on nitrogen transformation and sucralose (SUC) and acesulfame (ACE) degradation during SS composting. Results showed the cumulative rate of ACE and SUC in SS was ∼76 %. During SS composting, kaolin reduced NH3 emissions by 30.2–45.38 %, and N2O emissions by 38.4–38.9 %, while the Micro-KL and Nano-KL reduced nitrogen losses by 14.8 % and 12.5 %, respectively. Meanwhile, Micro-KL and Nano-KL increased ACE degradation by 76.8 % and 84.2 %, and SUC degradation by 75.3 % and 77.7 %, and significantly shifted microbial community structure. Furthermore, kaolin caused a positive association between Actinobacteria and sweetener degradation. Taken together, kaolin effectively inhibited nitrogen loss and promoted the degradation of ACE and SUC during the SS composting, which is of great significance for the removal of emerging organic pollutants in SS.

本研究调查了污水处理厂进水、出水和污水污泥(SS)中人工甜味剂的去向,并研究了微高岭土(Micro-KL)和纳米高岭土(Nano-KL)在 SS 堆肥过程中对氮转化以及三氯蔗糖(SUC)和安赛蜜(ACE)降解的影响。研究结果表明,蔗糖素(ACE)和安赛蜜(SUC)在 SS 中的累积降解率为 76%。在 SS 堆肥过程中,高岭土减少了 30.2%-45.38% 的 NH3 排放和 38.4%-38.9% 的 N2O 排放,而 Micro-KL 和 Nano-KL 则分别减少了 14.8% 和 12.5% 的氮损失。同时,微高岭土和纳米高岭土使 ACE 降解率分别提高了 76.8% 和 84.2%,SUC 降解率分别提高了 75.3% 和 77.7%,并显著改变了微生物群落结构。此外,高岭土还导致放线菌与甜味剂降解之间的正相关。综上所述,高岭土能有效抑制固体废弃物堆肥过程中氮的流失,促进 ACE 和 SUC 的降解,这对去除固体废弃物中新出现的有机污染物具有重要意义。
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引用次数: 0
Treatment of high ammonia anaerobically digested molasses wastewater using aerobic granular sludge reactor. 利用好氧颗粒污泥反应器处理高氨厌氧消化糖蜜废水。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-06-28 DOI: 10.1016/j.biortech.2024.131056
Xin Zou, Yiduo Yao, Mengjiao Gao, Yihui Zhang, Hengbo Guo, Yang Liu

This study addressed the treatment of high ammonia, low biodegradable chemical oxygen demand (bCOD) anaerobically digested molasses wastewater, utilizing an aerobic granular sludge (AGS) reactor. The AGS achieved 99 % ammonia removal regardless of the bCOD supplementation. By adding low ammonia (<60 mg/L), high bCOD raw molasses wastewater (before anaerobic digestion) as a carbon source, enhanced nitrogen removal, increasing from 10 % to 97 %, and improved sludge settleability via bio-induced calcite precipitation were observed. Functional genes prediction suggested two potential denitrification pathways, including heterotrophic denitrification by Paracoccus and Thauera, and autotrophic denitrification, specifically sulfide-oxidizing autotrophic denitrification by Thiobacillus. An increase in the relative abundance of microorganisms involved in heterotrophic denitrification was observed with the addition of high bCOD raw molasses wastewater. Consequently, incorporating raw molasses wastewater into the AGS presents a sustainable approach to achieve mixotrophic denitrification, maintain stable granular sludge and ensure stable treatment performance when treating anaerobically digested molasses wastewater.

本研究利用好氧颗粒污泥(AGS)反应器处理高氨氮、低生物降解化学需氧量(bCOD)的厌氧消化糖蜜废水。无论是否添加 bCOD,AGS 的氨氮去除率均达到 99%。通过添加低氨 (
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引用次数: 0
Diversity in mechanisms of natural humic acid enhanced current production in soil bioelectrochemical systems. 天然腐植酸增强土壤生物电化学系统电流产生机制的多样性。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-06-28 DOI: 10.1016/j.biortech.2024.131057
Xintong Gao, Kaixuan Liu, Chong Zhang, Xian Cao, Takashi Sakamakic, Xianning Li

The quinoid component of humic acids (HAs) had been studied as exogenous electron mediators (EMs), but the redox-mediating abilities of other functional groups remained unclear. This study evaluated the effects of various HAs functional groups on cellular respiration and extracellular electron transfer. The three EMs increased the current density compared to the control. Current density increased significantly after adding ultraviolet-irradiated HAs (UV-HAs), suggesting that nitrogenous group-mediated redox reactions contributed to high-density current generation. Structural equation model (SEM) results indicated that the contribution of nitrogen-containing groups to electron transfer could exceed 20%. This study proposed a synergistic mechanism: in the soil microbial fuel cells (soil-MFCs), HAs accelerated their component evolution through irreversible redox reactions and promoted extracellular electron transfer. Additionally, HAs-induced high expression of c-Cyts could further enhance high-density current generation. This study demonstrates that humic acids enhance electron transfer and current in bioelectrochemical systems, aiding sustainable energy optimization.

腐殖酸(HAs)中的醌类成分作为外源电子介质(EMs)已被研究过,但其他功能基团的氧化还原介导能力仍不清楚。本研究评估了各种腐殖酸功能团对细胞呼吸和细胞外电子传递的影响。与对照组相比,三种EM都能提高电流密度。加入紫外线照射过的 HAs(UV-HAs)后,电流密度明显增加,这表明氮基介导的氧化还原反应促成了高密度电流的产生。结构方程模型(SEM)结果表明,含氮基团对电子转移的贡献率可超过 20%。本研究提出了一种协同机制:在土壤微生物燃料电池(soil-MFCs)中,HAs 通过不可逆的氧化还原反应加速了其组分的演化,并促进了胞外电子传递。此外,腐殖酸诱导的 c-Cyts 高表达可进一步提高高密度电流的产生。这项研究表明,腐植酸能增强生物电化学系统中的电子传递和电流,有助于可持续能源优化。
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引用次数: 0
Innovative insights into organic nitrogen degradation through protein family domains analysis in chicken and pig manure composting using metagenomic sequencing 利用元基因组测序技术,通过分析鸡粪和猪粪堆肥中的蛋白质族域,对有机氮降解进行创新性研究。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-06-28 DOI: 10.1016/j.biortech.2024.131048
Lin Zhu , Caihong Huang , Lipin Li , Simiao Wang , Xinxin Wu , Guangchun Shan , Yu Tian

The nitrogen loss in composting is primarily driven by the transformation of organic nitrogen, yet the mechanisms underlying the degradation process remain incompletely understood. This study employed protein family domains (Pfams) analysis based on metagenomic sequencing to investigate the functional characteristics, key microorganisms, and environmental parameters influencing organic nitrogen degradation in chicken manure and pig manure composting. 154 Pfams associated with ammonification function were identified. Predominant Pfams: proteolytic peptidase, followed by chitin/cell wall degraders, least involved in nucleic acid degradation. Ammonifying microbial diversity was basically consistent among compost types, particularly in the thermophilic stage with the peak of abundance of dominant ammonifying microorganisms. Viruses played an important role in ammonification process, especially Uroviricota. 26 key ammonifying genera were identified by the microbial network. pH dominated the metabolic activity of ammonifying microorganisms in various manure compost types, primarily consisting of protein-degrading bacteria with stable community structures.

堆肥过程中的氮损失主要是由有机氮的转化所驱动的,但人们对降解过程的内在机制仍不完全了解。本研究利用基于元基因组测序的蛋白质家族结构域(Pfams)分析,研究了鸡粪和猪粪堆肥过程中影响有机氮降解的功能特征、关键微生物和环境参数。结果发现了 154 个与氨化功能相关的 Pfams。主要的 Pfams:蛋白水解肽酶,其次是几丁质/细胞壁降解剂,最少参与核酸降解。不同堆肥类型的氨化微生物多样性基本一致,尤其是在嗜热阶段,优势氨化微生物的丰度达到顶峰。病毒在氨化过程中发挥了重要作用,尤其是 Uroviricota。在不同类型的粪肥堆肥中,pH 值主导了氨化微生物的代谢活动,主要包括具有稳定群落结构的蛋白质降解菌。
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引用次数: 0
Principles, Challenges, and optimization of Indigenous Microalgae-Bacteria consortium for sustainable swine wastewater treatment. 用于可持续猪废水处理的本土微藻-细菌联合体的原理、挑战和优化。
IF 9.7 1区 环境科学与生态学 Q1 AGRICULTURAL ENGINEERING Pub Date : 2024-06-27 DOI: 10.1016/j.biortech.2024.131055
Sheng Yu, Zhipeng Chen, Mengting Li, Shuang Qiu, Zhe Lv, Shijian Ge

Indigenous microalgae-bacteria consortium (IMBC) offers significant advantages for swine wastewater (SW) treatment including enhanced adaptability and resource recovery. In this review, the approaches for enriching IMBC both in situ and ex situ were comprehensively described, followed by symbiotic mechanisms for IMBC which involve metabolic cross-feeding and signal transmission. Strategies for enhancing treatment efficiencies of SW-originated IMBC were then introduced, including improving SW quality, optimizing system operating conditions, and adjusting microbial activities. Recommendations for maximizing treatment efficiencies were particularly proposed using a decision tree approach. Moreover, removal/recovery mechanisms for typical pollutants in SW using IMBC were critically discussed. Ultimately, a technical route termed SW-IMBC-Crop-Pig was proposed, to achieve a closed-loop economy for pig farms by integrating SW treatment with crop cultivation. This review provides a deeper understanding of the mechanism and strategies for IMBC's resource recovery from SW.

本土微藻-细菌联合体(IMBC)在猪废水(SW)处理方面具有显著优势,包括增强适应性和资源回收。在本综述中,全面介绍了原位和非原位富集 IMBC 的方法,随后介绍了 IMBC 的共生机制,其中包括代谢交叉供养和信号传输。然后介绍了提高源自 SW 的 IMBC 处理效率的策略,包括改善 SW 质量、优化系统运行条件和调整微生物活动。特别是利用决策树方法提出了最大化处理效率的建议。此外,还批判性地讨论了利用 IMBC 去除/回收污水中典型污染物的机制。最终,提出了一条名为 "SW-IMBC-作物-猪 "的技术路线,通过将污水处理与作物栽培相结合,实现养猪场的闭环经济。本综述加深了人们对 IMBC 从 SW 中回收资源的机制和策略的理解。
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引用次数: 0
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Bioresource Technology
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