Biochemical Engineering Journal最新文献_第2页

Hairpin-locked modular polymerase/nickase cycle amplification enables ultrasensitive urinary miRNA detection for acute kidney injury diagnosis 发夹锁定模块化聚合酶/镍酶循环扩增使超灵敏的尿miRNA检测用于急性肾损伤诊断

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-02-03 DOI: 10.1016/j.bej.2026.110107

Keying Xu , Xinru Liu , Juan Zhu , Fenghe Li , Xiaoyan Liu , Jie Wang

The polymerase/nickase cycling amplification is a classic method for isothermal nucleic acid amplification, praised for its efficiency. However, its sensitivity and specificity are lacking for trace miRNA analysis. Herein, we have developed a novel modular polymerase/nickase cycling amplification technology (M-PNP) based on traditional polymerase/nickase probes (PNP), enhancing detection efficiency, sensitivity, and specificity. Specifically, M-PNP is formed by introducing a hairpin structure at the 5’ end of PNP and adding an auxiliary probe at the 3’ end. The hairpin structure acts like a lock, inhibiting non-specific nucleic acid amplification. In the presence of miRNA, the hairpin structure recognizes the miRNA and undergoes a conformational change, transforming into a new micro-hairpin structure. Through the action of polymerase and nickase, this transformation activates the polymerase/nickase cycling amplification reaction and promotes miRNA cycling, thereby enhancing detection sensitivity. The introduction of the auxiliary probe enables immediate activation of fluorescence signals during the target-triggered polymerase/nickase cycling amplification, significantly increasing detection speed and reducing time consumption. We used miR-21 from the urine of clinical kidney injury patients as the detection target and employed M-PNP for practical testing. The results showed that this platform can accurately distinguish kidney injury patients. This optimization and improvement injects new vitality and innovative ideas into the design of traditional polymerase/nickase cycling amplification probes.

聚合酶/镍酶循环扩增是一种经典的等温核酸扩增方法，因其高效而备受赞誉。然而，该方法在痕量miRNA分析中缺乏敏感性和特异性。在此，我们开发了一种基于传统聚合酶/镍酶探针（PNP）的新型模块化聚合酶/镍酶循环扩增技术（M-PNP），提高了检测效率、灵敏度和特异性。具体来说，M-PNP是通过在PNP的5 ‘端引入发夹结构，在3 ’端添加辅助探针形成的。发夹结构像锁一样，抑制非特异性核酸扩增。在miRNA存在的情况下，发夹结构识别miRNA并发生构象变化，转变为新的微发夹结构。这种转化通过聚合酶和镍酶的作用，激活聚合酶/镍酶循环扩增反应，促进miRNA循环，从而提高检测灵敏度。辅助探针的引入使荧光信号在目标触发的聚合酶/镍酶循环扩增过程中立即激活，显著提高了检测速度并减少了时间消耗。我们以临床肾损伤患者尿液中的miR-21作为检测靶点，并采用M-PNP进行实际检测。结果表明，该平台能够准确识别肾损伤患者。这种优化和改进为传统的聚合酶/镍酶循环扩增探针的设计注入了新的活力和创新思路。

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引用次数: 0

Platelet membrane-modified liquid metal nanoparticles enhanced cellular uptake and tumor photothermal therapy 血小板膜修饰的液态金属纳米颗粒增强细胞摄取和肿瘤光热治疗

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-02-03 DOI: 10.1016/j.bej.2026.110108

Siyuan Pang , Zhihang Wang , Yang Zou , Li Zhang , Yonggang Lv

Liquid metal (LM) nanoparticles have been widely used in photothermal therapy (PTT), but they are susceptible to oxidative inactivation and have poor targeting ability. Platelets (PLT) have many abundant membrane proteins on their surface that can be used to modify LM nanoparticles. Here, a gallium (Ga)-based LM-based nanoparticle delivery system was developed. The conducting polymer polypyrrole (PPy) was first grown in situ on the surface of LM nanoparticles by polymerization (named as LM@PPy). The oxidation resistance and photothermal stability of LM were improved. Subsequently, PLT membrane (PM) was extracted and coated on the surface of LM@PPy to prepare LM@PPy/PM. The antitumor effect of LM@PPy/PM was investigated through in vitro and in vivo experiments. It was demonstrated that the LM@PPy had better photothermal stability and their photothermal conversion efficiency reached 55 ± 2 %, which was higher than that of unmodified LM nanoparticles (31 ± 2 %). Most of the membrane proteins from PLT were retained on the prepared LM@PPy/PM. The PM coating effectively enhanced the tumor-targeting ability of the nanoparticles, leading to better tumor accumulation and antitumor effects in in vitro and in vivo. The findings showed that this nanoparticle delivery system provided a new technological solution to improve the antitumor ability of LM nanoparticles.

液态金属（LM）纳米颗粒在光热治疗（PTT）中得到了广泛的应用，但其易氧化失活且靶向能力差。血小板（PLT）表面有许多丰富的膜蛋白，可用于修饰LM纳米颗粒。本文开发了一种基于镓（Ga）的纳米颗粒递送系统。首先通过聚合在LM纳米颗粒表面原位生长导电聚合物聚吡咯（PPy）（命名为LM@PPy）。提高了LM的抗氧化性和光热稳定性。随后，提取PLT膜（PM），涂覆在LM@PPy表面，制备LM@PPy/PM。通过体外和体内实验考察LM@PPy/PM的抗肿瘤作用。结果表明，LM@PPy具有较好的光热稳定性，其光热转换效率达到55 ± 2 %，高于未修饰的LM纳米粒子（31 ± 2 %）。大部分来自PLT的膜蛋白保留在制备的LM@PPy/PM上。PM包被有效地增强了纳米颗粒的肿瘤靶向能力，从而在体外和体内具有更好的肿瘤蓄积和抗肿瘤作用。研究结果表明，该纳米颗粒递送系统为提高LM纳米颗粒的抗肿瘤能力提供了新的技术解决方案。

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引用次数: 0

A KPI-based experimental design strategy for bioprocess development 基于kpi的生物工艺开发实验设计策略

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-02-02 DOI: 10.1016/j.bej.2026.110096

Okyanus Yazgin , Martin F. Luna , Peter Neubauer , Ernesto C. Martinez , M. Nicolas Cruz Bournazou

Bioprocess development can benefit significantly from the use of mathematical models for prediction and optimization, yet the uncertainty in these models can hinder reliable early-stage decision-making for industrial-scale processes. This study introduces a telescopic model-based design of experiments approach that directly targets the reduction of uncertainty in key performance indicators (KPIs) at the optimum process conditions rather than focusing solely on model parameter precision. Using a sugarcane-to-ethanol biorefinery use case, the proposed approach is benchmarked against a traditional parameter-focused approach. Results demonstrate that the proposed strategy reduces KPI uncertainty more efficiently, identifies economically favorable process conditions faster, and prioritizes the estimation of parameters most influential on the KPI.

生物工艺开发可以从使用数学模型进行预测和优化中获益良多，但这些模型中的不确定性可能会阻碍工业规模工艺的可靠早期决策。本研究引入了一种基于伸缩模型的实验设计方法，该方法直接针对在最佳工艺条件下减少关键绩效指标（kpi）的不确定性，而不仅仅是关注模型参数的精度。使用甘蔗到乙醇的生物炼制用例，提出的方法与传统的以参数为中心的方法进行了基准测试。结果表明，该策略更有效地降低了KPI的不确定性，更快地识别经济上有利的工艺条件，并优先估计对KPI影响最大的参数。

引用次数: 0

Advanced bioelectronic scaffolds with electrostimulation for enhanced muscle regeneration 先进的生物电子支架与电刺激增强肌肉再生

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-02-02 DOI: 10.1016/j.bej.2026.110105

Xi Yang , Jianmin Wang , Liang Zhang , Fengling Zhuo , Linyan Ge , Liuhang Zhang , Jie Li , Fei Han , Guanghui Song , Xiaozhi Wang

Tissue engineering scaffolds are essential for facilitating tissue regeneration, and electrical stimulation has emerged as a powerful complementary strategy to accelerate this process. However, the functionality and performance of current scaffold systems remain suboptimal, limiting their therapeutic potential. In this study, a novel graphene-boron nitride-poly(lactic-co-glycolic acid) (GR-BN-PLGA) scaffold was fabricated using 3D printing technology for the repair of abdominal wall hernias in rats. The scaffold was constructed using two materials with distinct electrical properties, conductive GR-PLGA and insulating BN-PLGA, through a layered printing process. By integrating electrodes and microneedles, the scaffold was designed to establish a centrally directed electric field in the abdominal defect area, enabling effective electrical stimulation therapy. The results demonstrated that the combined application of the scaffold and electrical stimulation significantly upregulated the expression of α-smooth muscle actin, type I collagen, and Cell Proliferation Marker Protein Ki-67, thereby facilitating tissue remodeling. Meanwhile, the lower expression levels of proliferating cell nuclear antigen and connective tissue growth factor effectively suppressed excessive proliferation and fibrosis, aiding in the formation of stable and functional regenerated tissue. The synergistic application of conductive scaffolds and electrical stimulation provides a novel strategy for tissue repair and highlights its tremendous potential in accelerating tissue regeneration and promoting the formation of functional tissues.

组织工程支架对于促进组织再生至关重要，电刺激已成为加速这一过程的有力补充策略。然而，目前支架系统的功能和性能仍然不够理想，限制了它们的治疗潜力。本研究采用3D打印技术制备了新型石墨烯-氮化硼-聚乳酸-羟基乙酸（GR-BN-PLGA）支架，用于大鼠腹壁疝修复。支架采用导电GR-PLGA和绝缘BN-PLGA两种具有不同电性能的材料，通过分层印刷工艺构建而成。通过整合电极和微针，该支架被设计用于在腹部缺陷区域建立一个中心定向电场，从而实现有效的电刺激治疗。结果表明，支架与电刺激联合应用可显著上调α-平滑肌肌动蛋白、I型胶原和细胞增殖标记蛋白Ki-67的表达，促进组织重塑。同时，增殖细胞核抗原和结缔组织生长因子的低表达水平有效抑制了过度增殖和纤维化，有助于形成稳定和功能的再生组织。导电支架和电刺激的协同应用为组织修复提供了一种新的策略，并突出了其在加速组织再生和促进功能组织形成方面的巨大潜力。

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引用次数: 0

Urea-based treatment promotes the propagation of antibiotic resistance genes during sludge fermentation: Insights into its multifaceted roles in structure disruption, microbial community reshaping, and metabolic regulation 基于尿素的处理促进了污泥发酵过程中抗生素抗性基因的繁殖：深入了解其在结构破坏，微生物群落重塑和代谢调节中的多方面作用

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-01-30 DOI: 10.1016/j.bej.2026.110101

Minjun Zhao , Shuaijie Jin , Wenzhuo Li , Yuke Xu , Qin Zhang

Urea has been documented as an excellent promoter for improving sewage sludge (SS) fermentation, considering its effectiveness and economic feasibility, yet its effects on the fates of antibiotic resistance genes (ARGs) during this process are still unknown. Herein, the responses of ARGs distribution to urea exposure were studied, and the results revealed that urea exacerbated ARGs propagation, as evidenced by an increase of 66.8 % total abundance. Mechanistic exploration demonstrated that the presence of urea and free ammonia (FA) stripped the extracellular polymeric substances (EPS) and increased the cell membrane permeability, contributing to ARGs and mobile genetic elements (MGEs) release and consequently improved their horizontal transfer. Also, urea exhibited “screening effects” to enrich some harboring ARGs carriers (e.g., Bacteroidetes_norank, Tissierella and Firmicutes_norank). Further analysis found that the generated FA induced oxidative stress (e.g., katE and SOD1) and activated the SOS response (e.g., recA, recO, and recR), promoting ARGs formation, which could be further improved by unhydrolyzed urea through upregulating the metabolic functions (e.g., TCA cycle) associated with energy production. The structural equation model suggested that the upregulation of key metabolic pathways was the predominant contributor to the ARGs propagation. Collectively, this work explored the effects and underlying mechanisms of urea on ARGs' fates during SS fermentation, highlighting the potential environmental risks of urea-based treatment on resource recovery from SS.

考虑到其有效性和经济可行性，尿素已被证明是改善污水污泥（SS）发酵的良好促进剂，但在此过程中其对抗生素抗性基因（ARGs）命运的影响尚不清楚。研究了尿素胁迫下ARGs分布的变化规律，结果表明尿素胁迫下ARGs的繁殖速度加快，总丰度增加66.8% %。机制探索表明，尿素和游离氨（FA）的存在剥离了细胞外聚合物（EPS），增加了细胞膜的通透性，促进了ARGs和移动遗传元件（MGEs）的释放，从而促进了它们的水平转移。此外，尿素对一些携带ARGs的载体（如Bacteroidetes_norank、Tissierella和Firmicutes_norank）具有“筛选作用”。进一步分析发现，生成的FA诱导氧化应激（如katE和SOD1），激活SOS反应（如recA、recO和recR），促进ARGs的形成，而未水解尿素可通过上调与能量产生相关的代谢功能（如TCA循环）进一步改善ARGs的形成。结构方程模型表明，关键代谢途径的上调是ARGs繁殖的主要因素。总之，本研究探讨了尿素对SS发酵过程中ARGs命运的影响及其潜在机制，强调了尿素处理对SS资源回收的潜在环境风险。

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引用次数: 0

Comparative analysis of free and CLEA-immobilised α-amylase and α-glucosidase from Anoxybacillus flavithermus 2641 T 黄热无氧杆菌2641 游离α-淀粉酶和α-葡萄糖苷酶与固定化α-淀粉酶的比较分析

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-01-30 DOI: 10.1016/j.bej.2026.110106

Emel Alemdaroğlu , Fulya Ay , Dilşat Nigar Çolak , Halil İbrahim Güler , Ali Osman Belduz

α-Amylase (EC 3.2.1.1) and α-glucosidase (EC 3.2.1.20) are key enzymes in starch hydrolysis, widely applied in biotechnological and food industries. In this study, recombinant α-amylase (AflAmy) and α-glucosidase (AflGlu) from Anoxybacillus flavithermus 2641 ^T were purified using a cobalt affinity column, yielding proteins of approximately 50 kDa as confirmed by SDS-PAGE. Both enzymes were immobilised through the cross-linked enzyme aggregate (CLEA) method. Optimal CLEA preparation involved 96 % ammonium sulfate saturation at 4 °C for 30 min, followed by cross-linking with 5 mM glutaraldehyde for 2–3 h at room temperature. Free AflAmy exhibited optimal activity at pH 8.0 and 70 °C, while immobilization shifted its optimum to pH 9.0. Free AflGlu was most active at pH 8.0 and 60 °C, changing to 55 °C upon immobilization. CLEA forms displayed lower Km value for AflAmy-IM, indicating increased substrate affinity. Reusability tests showed immobilised AflAmy retained activity over nine cycles, AflGlu over three, and their combined form (Combi-CLEA) up to nine cycles. Thermal stability of immobilised AflAmy improved, maintaining activity for 150 min at 70 °C. Both free and immobilized forms of the enzymes achieved ∼50 % starch hydrolysis within 60 min, demonstrating comparable catalytic efficiency and enhanced operational stability after immobilization.

α-淀粉酶（EC 3.2.1.1）和α-葡萄糖苷酶（EC 3.2.1.20）是淀粉水解的关键酶，广泛应用于生物技术和食品工业。本研究利用钴亲和柱纯化了黄热无氧杆菌2641 T中的重组α-淀粉酶（AflAmy）和α-葡萄糖苷酶（AflGlu）， SDS-PAGE证实，所得蛋白约为50 kDa。两种酶通过交联酶聚集体（CLEA）法固定。最佳的CLEA制备方法是：96 %硫酸铵在4°C下饱和30 min，然后与5 mM戊二醛在室温下交联2-3 h。游离AflAmy在pH 8.0和70°C时表现出最佳活性，而固定化则在pH 9.0时表现出最佳活性。游离AflGlu在pH 8.0和60°C时最具活性，固定后为55°C。CLEA形式对AflAmy-IM显示较低的Km值，表明底物亲和力增加。可重用性测试表明，固定化AflAmy可保持9个循环的活性，AflGlu可保持3个循环，而它们的组合形式（Combi-CLEA）可保持9个循环。固定化AflAmy的热稳定性得到改善，在70°C下保持150 min的活性。游离形式和固定化形式的酶都在60 min内实现了~ 50% %的淀粉水解，显示出相当的催化效率和固定化后增强的操作稳定性。

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引用次数: 0

Enhanced anticancer activity through bidirectional fermentation of Epimedium brevicornum Maxim. with Morchella esculenta 短角淫羊藿双向发酵增强抗癌活性。羊肚菌

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-01-30 DOI: 10.1016/j.bej.2026.110103

Chengchuan Che , Lijun Sheng , Pingping Ding , Qi Liu , Huimin Zhao , Wenyu Han , Yunli Guo , Cuijuan Gao , Carol Sze Ki Lin , Xinxin Liang

Epimedium brevicornum Maxim. (EB), a traditional Chinese herbal medicine, exhibits notable anticancer and anti-aging activities attributed primarily to flavonoids such as epimedin C and icariin. However, its effective application is limited by low extraction efficiency and incomplete mechanistic understanding. Bidirectional fermentation is an effective microbial bioconversion strategy to enrich bioactive constituents or generate novel metabolites. Herein, the medicinal fungus Morchella esculenta (ME) was used to conduct bidirectional fermentation of EB. Compared with single-fermentation EB extract, the ME/EB extract had 2.39-fold and 1.44-fold greater epimedin C and icariin content, respectively. The ME/EB extract exhibited significant antiproliferative activity against cancer cells, with IC₅₀ values of 215 μg/mL for human cervical cancer HeLa cells and 235 μg/mL for non-small-cell lung cancer A549 cells. Moreover, the ME/EB extract induced cell-cycle arrest at G1/G0 phase in HeLa cells and S phase in A549 cells, while markedly suppressing cell migration and invasion. The ME/EB extract also down-regulated Bcl-2 protein expression and up-regulated Bax, PI3K, and Akt protein expression, thereby promoting cancer cell apoptosis. Therefore, bidirectional fermentation significantly enhanced EB anticancer activity, potentially through modulation of the PI3K/Akt signalling pathway. Overall, ME-mediated bidirectional fermentation of EB shows promise as a novel strategy for developing anticancer agents.

短角淫羊藿。中药叶黄素（EB）具有显著的抗肿瘤和抗衰老活性，其主要成分是淫羊藿苷和淫羊藿苷等黄酮类化合物。但萃取效率低，对其机理认识不全，限制了其有效应用。双向发酵是一种有效的微生物生物转化策略，可以丰富生物活性成分或产生新的代谢产物。本研究利用药用真菌羊肚菌（Morchella esculenta， ME）对EB进行双向发酵。与单发酵EB提取物相比，ME/EB提取物的淫羊藿苷C和淫羊藿苷含量分别提高了2.39倍和1.44倍。ME/EB提取物对癌细胞具有显著的抗增殖活性，对人宫颈癌HeLa细胞的IC₅0值为215 μg/mL，对非小细胞肺癌A549细胞的IC₅0值为235 μg/mL。此外，ME/EB提取物诱导HeLa细胞的G1/G0期和A549细胞的S期细胞周期停滞，同时显著抑制细胞迁移和侵袭。ME/EB提取物还能下调Bcl-2蛋白表达，上调Bax、PI3K和Akt蛋白表达，从而促进癌细胞凋亡。因此，双向发酵显著增强EB的抗癌活性，可能是通过调节PI3K/Akt信号通路。综上所述，me介导的EB双向发酵有望成为开发抗癌药物的新策略。

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引用次数: 0

A combination of domain fusion and homologous site transplantation improves the performance of Bst DNA polymerase in denaturation bubble-mediated strand exchange amplification 结构域融合和同源位点移植的结合提高了Bst DNA聚合酶在变性气泡介导的链交换扩增中的性能

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-01-30 DOI: 10.1016/j.bej.2026.110104

Quanling Dong , Qiming Chen , Keren Shang , Zhengrong Lu , Yuanlong Hu , Zhanmin Liu

Bst DNA polymerase has been widely used in isothermal nucleic-acid amplification platforms for pathogen detection and molecular diagnostics. Improving the catalytic efficiency and operational robustness of Bst DNA polymerase through protein engineering is therefore of substantial interest. In this study, we combined domain fusion with the introduction of a homologous site transplantation in the large fragment of Bst DNA polymerase (BstLF). Specifically, the double-stranded DNA-binding domain Sso7d was fused to BstLF via a flexible linker, while the F496H mutation was introduced. The resulting mutant, Sso7d-BstLF(F496H), demonstrated improved catalytic efficiency in isothermal amplification techniques of denaturation bubble-mediated strand exchange amplification (SEA). Compared with the wild-type, Sso7d-BstLF(F496H) reduced the SEA amplification time by approximately 50 %. In addition, the engineered polymerase retained robust amplification activity at 71°C and showed improved tolerance to pH fluctuations and to common inhibitory components (NaCl, EDTA, urea, ethanol, and SDS). Molecular docking and mutation energy calculations suggested that improved performance might be associated with additional hydrogen bonds between the R group and DNA, consistent with increased protein-DNA affinity. Molecular dynamics simulations further indicated that F496H preserves global structural stability while reducing local flexibility, providing a plausible structural basis for the observed activity enhancement. Collectively, these findings identify Sso7d–BstLF(F496H) as a promising polymerase for improved isothermal amplification–based molecular diagnostics.

Bst DNA聚合酶已广泛应用于病原体检测和分子诊断的等温核酸扩增平台。因此，通过蛋白质工程提高Bst DNA聚合酶的催化效率和操作稳健性具有重要意义。在这项研究中，我们将结构域融合与在Bst DNA聚合酶（BstLF）大片段中引入同源位点移植相结合。具体来说，双链dna结合域Sso7d通过一个柔性连接体融合到BstLF中，同时引入F496H突变。由此产生的突变体Sso7d-BstLF（F496H）在变性气泡介导链交换扩增（SEA）的等温扩增技术中表现出更高的催化效率。与野生型相比，Sso7d-BstLF（F496H）将SEA扩增时间缩短了约50% %。此外，工程聚合酶在71°C下保持了强大的扩增活性，并表现出对pH波动和常见抑制成分（NaCl， EDTA，尿素，乙醇和SDS）的耐受性增强。分子对接和突变能量计算表明，性能的提高可能与R基团和DNA之间额外的氢键有关，这与蛋白质-DNA亲和力的增加相一致。分子动力学模拟进一步表明，F496H保持了整体结构稳定性，同时降低了局部柔韧性，为观察到的活性增强提供了合理的结构基础。总的来说，这些发现确定Sso7d-BstLF （F496H）是一种有前途的聚合酶，用于改进等温扩增的分子诊断。

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引用次数: 0

Enhanced expression and purification strategy for β2-microglobulin in Escherichia coli β2微球蛋白在大肠杆菌中的增强表达及纯化策略

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-01-30 DOI: 10.1016/j.bej.2026.110100

Boram Kim , Junwoo Hwang , Dong-Hyun Seo , Sun Taek Kim , Ji-Hun Kim , Kyoung-Seok Ryu

β2-microglobulin (β2m) is a biomarker for various renal diseases and forms the major histocompatibility complex class I (MHC-I) with the heavy chain. The presenting antigenic peptide on the MHC-I is critical for the cytotoxic T-cell mediated immune responses, and thus extensive efforts have been paid to the in vitro reconstitution of the MHC-I through co-refolding of the heavy chain and β2m. Although β2m that contains a single disulfide bond is highly stable, it is typically expressed as an insoluble form in Escherichia coli (E. coli). In this study, native β2m was prepared via soluble expression in the E. coli SHuffle T7 Express strain, achieving a yield of 30–40 mg per liter of Luria-Bertani (LB) medium, more than 10-fold higher than that obtained through periplasmic expression. The refolding of the heavy chain with the purified β2m successfully produced the cognate peptide-loaded MHC-I (pMHC-I). The SHuffle T7 system offers a straightforward approach for producing functional β2m for future researches in immunology and structural biology.

β2-微球蛋白（β2m）是多种肾脏疾病的生物标志物，与重链形成主要的组织相容性复合体I类（MHC-I）。MHC-I上的递呈抗原肽对于细胞毒性t细胞介导的免疫应答至关重要，因此通过重链和β2m的共重折叠来体外重建MHC-I已经付出了大量的努力。虽然含有单键二硫键的β2m高度稳定，但它在大肠杆菌（E. coli）中通常以不溶形式表达。本研究采用可溶性表达法在大肠杆菌SHuffle T7 Express菌株中制备了天然β2m，在LB培养基中获得了30-40 mg / l的产量，比采用周质表达法获得的产量高出10倍以上。重链与纯化的β2m重折叠成功地产生同源肽装载MHC-I （pMHC-I）。SHuffle T7系统为免疫学和结构生物学的未来研究提供了一种直接产生功能性β2m的方法。

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引用次数: 0

Nitrogen removal performance and low-temperature impact of hybrid membrane aerated biofilms reactor (H-MABR) 混合膜曝气生物膜反应器（H-MABR）脱氮性能及低温影响

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2026-01-29 DOI: 10.1016/j.bej.2026.110102

Youzhao Wang , Jie Han , Mingdong Chang , Yongguang Ma , Xiaoyan Dang , Shumin He , Zhipeng Wang , Rongxiao Zhang , Junnan Liu , Jinxiang Wang , Lin Zhai , Junting Wang , Zhenning Lv , Tong Zhu

Declining microbial activity under low-temperature conditions poses a persistent challenge for biological wastewater treatment. Conventional Membrane Aerated Biofilm Reactor (C-MABR) has the advantages of high mass transfer efficiency, stable treatment effect and low aeration energy consumption. However, treatment capacity of C-MABR is significantly compromised under low-temperature conditions. In this work, a novel hybrid membrane aerated biofilm reactor (H-MABR) was developed by integrating biomass carriers with membrane aeration to enhance system robustness in cold conditions. Performance, process optimization, and microbial community characteristics of H‑MABR were systematically evaluated at

10 \sim 12 ℃

. The results indicated that under approximately 200 mg/L of COD, approximately 50 mg/L of

{NH}_{4}^{+} - N

and 12 h of HRT, COD removal loading rates averaged 0.244 kg/(m³∙d),

{NH}_{4}^{+} - N

removal loading rates averaged 0.051 kg N/(m³∙d) and TN removal loading rates averaged 0.038 kg N/(m³∙d), respectively. Using response surface methodology (RSM), optimal operating window for pollutant removal was identified as followed: temperature

10 \sim 12 ℃

, pH 7.5, C/N ratio 3.71, and aeration pressure 0.02 MPa. Microbial community analysis demonstrated that exposure to low temperature markedly reshaped the community structure in H‑MABR, indicating a strong linkage between temperature and functional microbial community. Overall, this study elucidated impact of low-temperature operation on H‑MABR performance and microbial ecology and provided practical insights to guide the design and operation for wastewater treatment in cold environments.

低温条件下微生物活性下降是废水生物处理的一个长期挑战。常规膜曝气生物膜反应器（C-MABR）具有传质效率高、处理效果稳定、曝气能耗低等优点。然而，在低温条件下，C-MABR的处理能力明显受损。在这项工作中，通过将生物质载体与膜曝气相结合，开发了一种新型混合膜曝气生物膜反应器（H-MABR），以提高系统在寒冷条件下的稳健性。在10 ~ 12℃条件下对H‑MABR的性能、工艺优化和微生物群落特征进行了系统评价。结果表明，在约200 mg/L COD、约50 mg/L NH4+−N和12 h HRT条件下，COD去除率平均为0.244 kg/(m³∙d)， NH4+−N去除率平均为0.051 kg N/(m³∙d)， TN去除率平均为0.038 kg N/（m³∙d）。利用响应面法（RSM）确定了污染物去除的最佳操作窗口：温度10 ~ 12℃，pH值7.5，C/N比3.71，曝气压力0.02 MPa。微生物群落分析表明，低温显著重塑了H - MABR的群落结构，表明温度与功能微生物群落之间存在很强的联系。总体而言，本研究阐明了低温运行对H - MABR性能和微生物生态的影响，为指导低温环境下废水处理的设计和运行提供了实践见解。

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