Journal of biotechnology最新文献_第4页

Optical, structural, and antifungal properties of nanosilver borate bioactive glass synthesized using gamma rays on the survival of Candida albicans and Candida tropicalis 利用伽马射线合成的纳米硼酸银生物活性玻璃对白色念珠菌和热带念珠菌生存的光学、结构和抗真菌特性。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-21 DOI: 10.1016/j.jbiotec.2025.11.013

E.M. Abou Hussein , Rasha Mohammad Fathy

A New glass system with a composition of; 50.3 B₂O₃+ 21 CaO + 8 MgO + 8 K₂O + 8 Na₂O + 4 P₂O₅ + 0.7 Nano Ag₂O, was prepared by the traditional melting technique. Some of its structural, optical, and physical characteristics were investigated regarding the effect of gamma radiation. The XRD pattern revealed the amorphous nature of the fabricated glass sample. UV–visible absorption spectra revealed an obvious peak before 400 nm due to the absorption of Ag^2 + ions, and a photo-reduction reaction to Ag⁰ was expected due to the impact of gamma irradiation. FTIR absorption spectra displayed structural bands in the region 800–1600 cm⁻¹ recognized to the specific vibrations of BO₃, BO₄ and PO₂. Results displayed optical and structural stability against gamma irradiation doses of 25 and 50 kGy, and revealed a positive effect of irradiation to enhance the antifungal efficiency of the glass or in sterilization processes before usage. The antifungal activity of borate bioglass doped with silver nanoparticles (BG/Ag) against Candida albicans and Candida tropicalis was assessed. BG/Ag was mostly effective against C. albicans. Furthermore, at a dosage of 40.0 mg/mL, BG/Ag considerably reduced biofilm formation of C. albicans and C. tropicalis (90.79 and 75.07 %). At 40 mg/mL, BG/Ag significantly reduced Candida viability, colony counts, and phospholipase activity. BG/Ag can be successfully applied as an attractive biomaterial for the medication of serious fungal infections. The novelty in the study is the synthesis of borate bioactive glass containing nanosilver- and the use of gamma irradiation to improve its biological properties.

一种新型玻璃体系，其组成为；采用传统熔融法制备了50.3 B2O3+ 21 CaO + 8 MgO + 8 K2O + 8 Na2O+ 4 P2O5 + 0.7纳米Ag2O。在伽玛辐射的影响下，研究了它的一些结构、光学和物理特性。XRD图显示了制备的玻璃样品的非晶态性质。由于Ag2+离子的吸收，紫外可见吸收光谱在400nm前出现明显的峰值，并且由于伽马辐照的影响，预计会发生Ag0的光还原反应。FTIR吸收光谱显示出800-1600cm-1区域的结构带，这是BO3、BO4和PO2特有的振动特征。结果表明，在25和50 kGy的伽马辐射剂量下，玻璃具有光学稳定性和结构稳定性，并显示辐照对提高玻璃的抗真菌效率或在使用前的灭菌过程中具有积极作用。研究了纳米银掺杂硼酸盐生物玻璃对白色念珠菌和热带念珠菌的抑菌活性。BG/Ag对白色念珠菌最有效。此外，在40.0mg/mL的剂量下，BG/Ag显著减少了白色念珠菌和热带念珠菌的生物膜形成（90.79%和75.07%）。在40mg/mL浓度下，BG/Ag显著降低念珠菌活力、菌落计数和磷脂酶活性。BG/Ag可以作为一种有吸引力的生物材料成功应用于严重真菌感染的治疗。这项研究的新颖之处在于合成了含有纳米银的硼酸盐生物活性玻璃，并使用伽马射线照射来改善其生物特性。

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引用次数: 0

Halotolerant black yeast Neophaeotheca triangularis as a source of melanin 耐盐黑酵母菌，作为黑色素的来源。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-12 DOI: 10.1016/j.jbiotec.2025.11.011

Mercedes Peña , Patricia Gómez-Villegas , Jose Prados , Consolación Melguizo , Carla C.C.R. de Carvalho

Archaea and bacteria are the most studied extremophiles, but fungi also demonstrate remarkable tolerance, particularly in hypersaline environments such as solar salterns. Among salt-adapted fungi, black yeasts have been shown to be adapted to such environments, having developed defense mechanisms such as the production of melanin, a pigment that plays a crucial role in environmental stress protection. Melanin is a complex, high-molecular-weight polymer widely found across biological kingdoms. During the isolation of microorganisms from samples collected in crystallization ponds in a saltern, a black yeast was found able to produce significant amounts of melanin. The yeast was identified as belonging to the species Neophaeotheca triangularis. The aim of this study was to optimize the cultivation conditions of the strain, to characterize the produced melanin, and to assess its biological activities, including its antitumor and antioxidant properties.

古细菌和细菌是研究最多的极端微生物，但真菌也表现出非凡的耐受性，特别是在高盐环境中，如太阳盐沼。在适应盐的真菌中，黑酵母已经被证明能够适应这样的环境，已经发展出防御机制，如产生黑色素，一种在环境应激保护中起关键作用的色素。黑色素是一种复杂的高分子量聚合物，广泛存在于生物王国中。在从盐沼结晶池中收集的样品中分离微生物时，发现一种黑酵母能够产生大量的黑色素。经鉴定，该酵母菌属Neophaeotheca triangularis。本研究的目的是优化菌株的培养条件，表征所产生的黑色素，并评估其生物活性，包括抗肿瘤和抗氧化性能。

引用次数: 0

FRET two-hybrid assay-based target drug screening in living cells 基于FRET双杂交分析的活细胞靶标药物筛选。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-12 DOI: 10.1016/j.jbiotec.2025.11.010

Beini Sun , Gengqiang Cao , Tian Gan , Yue Wang , Rumeng Qu , Lin Hu , Yuan Chang , Min Hu , Xiaoping Wang , Tongsheng Chen

Precise targeted drug screening is the key to improve the efficiency of tumour targeted therapy. This report presents a live cell FRET two-hybrid assay-based targeted drug screening method (FRET-HBTDS). In FRET-HBTDS, the cells co-expressing donor- and acceptor-labelled targets were cultured in 96-well plates, quantitative FRET imaging was then performed on a self-built automated FRET microscope (FRETscope) well-by-well, and FRET two-hybrid assay was used to obtain the maximum donor-centre FRET efficiency (E_Dmax), maximum acceptor-centre FRET efficiency (E_Amax) and stoichiometric ratios (N_A/N_D). The FRET-HBTDS method was performed on the FRETscope with a 20 × objective for the cells co-expressing CFP-Bcl-xL and YFP-Bak to assess the action of eight compounds (A1331852, S63845, AC, DSF/Cu, Met, REGO, SOFA, ABT199) on the interaction between Bcl-xL and Bak. After 7 h of treatment with these compounds respectively, only the A1331852 group showed significantly lower E_Dmax and E_Amax compared to the control group, and a significant increase in N_A/N_D, suggesting that A1331852 unlocks the direct interaction between Bcl-xL and Bak, thus releasing Bak to induce cell death. In addition, the N_A/N_D of the DSF/Cu group was significantly higher than of the control group, suggesting that DSF/Cu altered the stoichiometry of the Bcl-xL-Bak complex. Our data firmly demonstrate that A1331852 unlocks the binding state of Bcl-xL and Bak, while DSF/Cu modifies the structure of the Bcl-xL-Bak complex. These findings demonstrate that FRET-HBTDS can be used to assess the efficacy of a drug by revealing the binding state and complex molecular structure of the target proteins using FRET technology in living cells, which may be a potential targeted drug screening method.

精确的靶向药物筛选是提高肿瘤靶向治疗效率的关键。本报告提出了一种基于活细胞FRET双杂交测定的靶向药物筛选方法（FRET- hbtds）。在FRET- hbtds中，将共表达供体和受体标记靶标的细胞培养在96孔板中，然后在自建的自动FRET显微镜（FRETscope）上逐孔进行定量FRET成像，并使用FRET双杂交法获得最大供体中心FRET效率（EDmax），最大受体中心FRET效率（EAmax）和化学计量比（NA/ND）。在FRETscope上对共表达CFP-Bcl-xL和YFP-Bak的细胞采用FRET-HBTDS法，以20倍的倍率评估8个化合物（A1331852、S63845、AC、DSF/Cu、Met、REGO、SOFA、ABT199）对Bcl-xL和Bak相互作用的影响。分别用这些化合物处理7h后，只有A1331852组EDmax和EAmax较对照组显著降低，NA/ND显著升高，提示A1331852开启了Bcl-xL与Bak的直接相互作用，从而释放Bak诱导细胞死亡。此外，DSF/Cu组的NA/ND显著高于对照组，表明DSF/Cu改变了Bcl-xL-Bak复合物的化学计量。我们的数据有力地证明了A1331852解锁了Bcl-xL和Bak的结合状态，而DSF/Cu修饰了Bcl-xL-Bak复合物的结构。这些发现表明，FRET- hbtds可以利用FRET技术在活细胞中揭示靶蛋白的结合状态和复杂的分子结构，从而评估药物的疗效，这可能是一种潜在的靶向药物筛选方法。

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引用次数: 0

Corrigendum to “Substitution of reactive centre loop residues from C1 esterase inhibitor increases the inhibitory specificity of alpha-1 antitrypsin for plasma kallikrein” [J. Biotechnol. 405 (2025) 205–214] 从C1酯酶抑制剂中取代反应性中心环残基增加α -1抗胰蛋白酶对血浆钾激肽的抑制特异性[J]。生物工程学报，2005(5):389 - 389。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-08 DOI: 10.1016/j.jbiotec.2025.11.003

Sangavi Sivananthan , S. Ameer Ahmed , Ammaar M. Baig , Varsha Bhakta , William P. Sheffield

引用次数: 0

Ternary deep eutectic solvent (T-CuCl2-EG) for lignin removal from corn straw 三元深共熔溶剂（T-CuCl2-EG）脱除玉米秸秆木质素。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-07 DOI: 10.1016/j.jbiotec.2025.11.009

Kang Zhao , Sha-Sha Zhao , Quan-Wei Liu , Zhen Liu

To address the challenge of lignin removal while preserving cellulose in lignocellulosic biomass valorization, this study developed a novel ternary deep eutectic solvent (DES) system composed of triethylbenzyl ammonium chloride (T, as hydrogen bond acceptor), copper chloride (Lewis acid catalyst), and ethylene glycol (EG, hydrogen bond donor) for microwave-assisted pretreatment of corn straw. The DES was designed to leverage synergistic effects between Lewis acid catalysis and hydrogen bond disruption for selective lignin extraction. Optimization via Box-Behnken experimental design revealed that optimal delignification (96.52 %) and hemicellulose removal (94.60 %) with 94.79 % cellulose retention were achieved under microwave conditions of 130°C, 40 min, liquid-solid ratio 13.1, and CuCl₂/T molar ratio 0.198. Comparative characterization of raw and pretreated biomass using SEM, FT-IR, and XRD demonstrated that T-CuCl₂-EG disrupted the lignocellulose matrix by breaking ether/ester bonds in lignin-carbohydrate complexes, selectively solubilizing lignin/hemicellulose while preserving cellulose crystallinity. This work provides a sustainable and efficient strategy for biomass fractionation, with potential applications in biorefineries.

为了解决木质素脱除的挑战，同时保留木质纤维素生物质的价值，本研究开发了一种新型的三元深共晶溶剂（DES）体系，该体系由三乙基苄基氯化铵（T，作为氢键受体）、氯化铜（Lewis酸催化剂）和乙二醇（EG，氢键供体）组成，用于微波辅助预处理玉米秸秆。DES旨在利用Lewis酸催化和氢键破坏之间的协同效应来选择性提取木质素。Box-Behnken实验优化结果表明，在130℃、40min、液固比13.1、CuCl₂/T摩尔比0.198的微波条件下，脱木质素率为96.52%，半纤维素去除率为94.60%，纤维素保留率为94.79%。利用SEM、FT-IR和XRD对未处理和预处理的生物质进行对比表征，结果表明，T-CuCl 2 -EG通过破坏木质素-碳水化合物复合物中的醚/酯键来破坏木质纤维素基质，在保持纤维素结晶度的同时选择性地溶解木质素/半纤维素。这项工作为生物质分馏提供了一种可持续和有效的策略，在生物炼制中具有潜在的应用前景。

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引用次数: 0

Differential metabolic reprogramming due to nitrogen starvation leads to remobilization of storage metabolites in microalgal strains 氮饥饿引起的差异代谢重编程导致微藻菌株储存代谢物的重新动员。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-07 DOI: 10.1016/j.jbiotec.2025.11.008

Lidiane Covell , Mariana Machado , Marcelo Gomes Marçal Vieira Vaz , Auxiliadora Oliveira Martins , Carolina Cassano Monte Bello , Umarah Mubeen , Dilson Novais Rocha , Marcio Arêdes Martins , Camila Caldana , Wagner L. Araújo , Adriano Nunes-Nesi

Microalgal stress-related responses to nitrogen (N) deprivation are determined by genetically defined metabolic changes, which are further influenced by cultivation and nutritional conditions. In this study, we evaluated the metabolic responses of five chlorophyte strains to varying N concentrations. The strains included Chlamydomonas reinhardtii and Scenedesmus obliquus, which accumulated higher levels of starch, and Monoraphidium sp., Chlamydomonas sp., and Chlorella vulgaris, characterized by differences in both the levels and compositions of accumulated lipids. Under N-starved condition, all strains exhibited increased starch and neutral lipid content, with strain-specific variations. In this treatment, the content of branched-chain amino acids decreased in Monoraphidium sp., likely due to amino acid catabolism associated with the autophagy and triacylglycerol (TAG) accumulation. Furthermore, we observed a remodeling of lipid classes in S. obliquus, C. vulgaris, Chlamydomonas sp., and Monoraphidium sp., including reductions in digalactosyl diacylglycerol, monogalactosyl diacylglycerol, and sulfoquinovosyl diacylglycerol, accompanied by an increase in TAGs. Under N-starvation several fatty acids of C. vulgaris, Chlamydomonas sp., and C reinhardtii, including α-linolenic acid (C18:3), oleic acid (C18:1), and linoleic acid (C18:2) increase. In contrast, Monoraphidium sp. accumulated long-chain fatty acids such as lumequic acid (C30:1), nervonic acid (C24:1), docosanoic acid (C22:0), and erucic acid (C22:1). Collectively, these results indicate that chlorphyte strains exhibit distinct metabolic responses to N depletion in the medium. These findings highlight the importance of strain-specific assessments when developing the N management strategies for microalgal cultivation and biotechnological applications.

微藻对氮(N)剥夺的应激相关反应是由遗传上确定的代谢变化决定的，这些变化进一步受到培养和营养条件的影响。在本研究中，我们评估了五种绿藻菌株对不同氮浓度的代谢反应。其中，莱茵衣单胞菌（Chlamydomonas reinhardtii）和斜状衣单胞菌（Scenedesmus obliquus）淀粉积累水平较高，单胞菌（Monoraphidium sp.）、衣单胞菌（Chlamydomonas sp.）和普通小球藻（Chlorella vulgaris）的脂肪积累水平和组成存在差异。在缺氮条件下，所有菌株的淀粉和中性脂含量均有所增加，但存在菌株特异性差异。在这种处理下，单霉的支链氨基酸含量下降，可能是由于氨基酸分解代谢与自噬和三酰甘油（TAG）积累有关。此外，我们还观察到斜螺旋藻、C. vulgaris、衣藻和单胞菌中脂类的重塑，包括双半乳糖二酰基甘油、单半乳糖二酰基甘油和磺基喹啉二酰基甘油的减少，同时伴随着标签的增加。在缺氮条件下，C. vulgaris、Chlamydomonas sp.和C. reinhardtii的α-亚麻酸（C18:3）、油酸（C18:1）和亚油酸（C18:2）等脂肪酸含量增加。相比之下，单紫藓属积累了长链脂肪酸，如光酸（C30:1）、神经酸（C24:1）、二十二酸（C22:0）和芥酸（C22:1）。综上所述，这些结果表明，绿藻菌株对培养基中氮的消耗表现出明显的代谢反应。这些发现强调了在制定微藻培养和生物技术应用的氮管理策略时，菌株特异性评估的重要性。

{"title":"Differential metabolic reprogramming due to nitrogen starvation leads to remobilization of storage metabolites in microalgal strains","authors":"Lidiane Covell , Mariana Machado , Marcelo Gomes Marçal Vieira Vaz , Auxiliadora Oliveira Martins , Carolina Cassano Monte Bello , Umarah Mubeen , Dilson Novais Rocha , Marcio Arêdes Martins , Camila Caldana , Wagner L. Araújo , Adriano Nunes-Nesi","doi":"10.1016/j.jbiotec.2025.11.008","DOIUrl":"10.1016/j.jbiotec.2025.11.008","url":null,"abstract":"<div><div>Microalgal stress-related responses to nitrogen (N) deprivation are determined by genetically defined metabolic changes, which are further influenced by cultivation and nutritional conditions. In this study, we evaluated the metabolic responses of five chlorophyte strains to varying N concentrations. The strains included Chlamydomonas reinhardtii and Scenedesmus obliquus, which accumulated higher levels of starch, and Monoraphidium sp., Chlamydomonas sp., and Chlorella vulgaris, characterized by differences in both the levels and compositions of accumulated lipids. Under N-starved condition, all strains exhibited increased starch and neutral lipid content, with strain-specific variations. In this treatment, the content of branched-chain amino acids decreased in Monoraphidium sp., likely due to amino acid catabolism associated with the autophagy and triacylglycerol (TAG) accumulation. Furthermore, we observed a remodeling of lipid classes in S. obliquus, C. vulgaris, Chlamydomonas sp., and Monoraphidium sp., including reductions in digalactosyl diacylglycerol, monogalactosyl diacylglycerol, and sulfoquinovosyl diacylglycerol, accompanied by an increase in TAGs. Under N-starvation several fatty acids of C. vulgaris, Chlamydomonas sp., and C reinhardtii, including α-linolenic acid (C18:3), oleic acid (C18:1), and linoleic acid (C18:2) increase. In contrast, Monoraphidium sp. accumulated long-chain fatty acids such as lumequic acid (C30:1), nervonic acid (C24:1), docosanoic acid (C22:0), and erucic acid (C22:1). Collectively, these results indicate that chlorphyte strains exhibit distinct metabolic responses to N depletion in the medium. These findings highlight the importance of strain-specific assessments when developing the N management strategies for microalgal cultivation and biotechnological applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 231-246"},"PeriodicalIF":3.9,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145482251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High temperature drives torularhodin-dominant carotenoid overaccumulation in Rhodosporidiobolus odoratus XQR via coordinated transcriptomic–metabolomic reprogramming 高温通过协调的转录组-代谢组重编程驱动桃红孢子虫XQR中以环孢素为主的类胡萝卜素过度积累。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-06 DOI: 10.1016/j.jbiotec.2025.11.001

Die Zhao , Chunji Li , Nan Zeng , Dandan Wang , Anqi Zeng , Guohui Yu , Ning Zhang , Bingxue Li

Carotenoids are bioactive pigments widely used in food, pharmaceutical, and cosmetic industries. Rhodosporidiobolus odoratus co-produces commercially significant carotenoids, including β-carotene, torulene, and torularhodin. This study aimed to reveal how carotenoid biosynthesis in R. odoratus XQR, optimally grown at 20 °C, responds to low (10 °C) and high (30 °C) temperatures. After five days, total carotenoids reached 118.55 μg/g DCW and 0.40 μg/mL at 30 °C, ∼3-fold higher than the 20 °C control (41.25 μg/g DCW and 0.14 μg/mL). Torularhodin showed a pronounced increase at 30 °C (37.26 μg/g DCW and 0.12 μg/mL), ∼9-fold above the control (4.06 μg/g DCW and 0.014 μg/mL). At 10 °C, total carotenoids declined to 26.09 μg/g DCW, with a slight, non-significant rise in volumetric titer to 0.17 μg/mL. High temperature elevated reactive oxygen species (ROS) and superoxide dismutase (SOD) activity while reducing total protein, whereas low temperature maintained stable ROS, induced moderate SOD, and caused protein decline; catalase (CAT) activity changed minimally under both conditions. Our integrated data suggest that high temperature promotes carotenoid overaccumulation through upregulation of key terpenoid backbone biosynthetic genes (HMGCS, hmgA, and mvaD) and carotenogenic genes (crtYB, crtI, crtZ, and crtA), coupled with a metabolic shift that enhanced precursor supply. These coordinated responses explain torularhodin-dominant accumulation and differ from related yeasts, suggesting species-specific regulation. This work provides new mechanistic insights into temperature-driven carotenogenesis in R. odoratus XQR and highlights targets for metabolic engineering.

类胡萝卜素是一种生物活性色素，广泛应用于食品、医药、化妆品等行业。odoratus Rhodosporidiobolus共同生产具有商业意义的类胡萝卜素，包括β-胡萝卜素、托鲁烯和托鲁霍丁。本研究旨在揭示在20°C条件下生长的R. odoratus XQR的类胡萝卜素生物合成对低（10°C）和高（30°C）温度的响应。5 d后，30 ℃条件下，类胡萝卜素总量分别达到118.55μg/g DCW和0.40μg/mL，是20 ℃对照（41.25μg/g DCW和0.14μg/mL）的3倍左右。Torularhodin在30 °C时（37.26μg/g DCW和0.12μg/mL）显著增加，是对照组（4.06μg/g DCW和0.014μg/mL）的9倍。在10 °C时，总类胡萝卜素下降到26.09μg/g DCW，体积滴度略有上升，达到0.17μg/mL。高温升高活性氧（ROS）和超氧化物歧化酶（SOD）活性，降低总蛋白，低温维持稳定的ROS，诱导适度的SOD，导致蛋白下降；过氧化氢酶（CAT）活性在两种条件下变化最小。我们的综合数据表明，高温通过上调关键萜类主干生物合成基因（HMGCS、hmgA和mvaD）和致胡萝卜素基因（crtYB、crtI、crtZ和crtA），以及代谢转变促进了前体供应，从而促进了类胡萝卜素的过度积累。这些协调的反应解释了torularhodin显性积累，并与相关酵母不同，表明物种特异性调节。这项工作为温度驱动的胡萝卜素生成提供了新的机制见解，并突出了代谢工程的靶点。

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引用次数: 0

Metabolic engineering of carotenoid biosynthesis in Yarrowia lipolytica 脂溶耶氏菌类胡萝卜素生物合成的代谢工程。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-05 DOI: 10.1016/j.jbiotec.2025.11.005

Shuyao Ma , Yufan Jing , Fangyuan Liu , Xiaolong Zheng , Xing Zhang , Meng Qiao

As a non-conventional oleaginous yeast, Yarrowia lipolytica (Y. lipolytica) has emerged as a promising host for carotenoid production due to its robust metabolism capabilities and lipids accumulate ability. The metabolic engineering of Y. lipolytica necessitates genetic expression tools, genome editing technologies, and systematic strategies for optimizing the global biosynthesis pathway. In this review, we summarize key genetic regulatory elements and gene editing tools that have been employed to enhance carotenoid biosynthesis in Y. lipolytica. We also examine representative metabolic engineering cases involving the production of carotenoids, such as lycopene, β-carotene, astaxanthin, and lutein, by Y. lipolytica with a focus on precursor pathway enhancement, subcellular compartmentalization, as well as enzyme modification and expression tuning. Furthermore, perspectives are provided on future strategies to engineer high-performance Y. lipolytica strains for industrial carotenoid production.

作为一种非常规的产油酵母，聚脂耶氏酵母（Y. lipolytica）由于其强大的代谢能力和脂质积累能力而成为类胡萝卜素生产的有希望的宿主。脂质体酵母的代谢工程需要基因表达工具、基因组编辑技术和优化全局生物合成途径的系统策略。在这篇综述中，我们总结了关键的遗传调控元件和基因编辑工具，已被用来促进类胡萝卜素的生物合成。我们还研究了具有代表性的代谢工程案例，涉及类胡萝卜素的产生，如番茄红素、β-胡萝卜素、虾青素和叶黄素，通过脂质体Y.脂质体，重点关注前体途径增强、亚细胞区隔化以及酶修饰和表达调节。此外，展望了未来的策略，以工程高性能的脂质体芽孢杆菌菌株的工业类胡萝卜素生产。

引用次数: 0

Increased purity and refolding yield of bacterial inclusion bodies by recursive high pressure homogenization 通过递推高压均质提高细菌包涵体的纯度和再折叠率。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-05 DOI: 10.1016/j.jbiotec.2025.11.004

Robert Klausser , Lukas Veiter , Julian Kopp , Nikolaus Hammerschmidt , Tiffany Frierss , Florian Gisperg , Mohamed Elshazly , Eva Prada Brichtova , Michael Martinetz , Martin Voigtmann , Oliver Spadiut

Therapeutic proteins are often produced intracellularly in the bacterium Escherichia coli in aggregated form, known as inclusion bodies, which must be solubilized and refolded to recover the biologically active product. However, the industrially established refolding method of batch dilution is still carried out at low protein concentrations, leading to high water consumption, large equipment footprints, and poor sustainability. An underexplored way to increase refolding yields and thereby enable higher product concentrations after refolding is the optimization of cell lysis and inclusion body washing. In this study, we developed and evaluated a “recursive high pressure homogenization” approach, wherein wash steps and additional homogenization cycles were combined to enhance the final purity of inclusion bodies before solubilization. The degree of cell lysis, nucleic acid release and final inclusion body purity were compared to conventional “linear washing”, where all homogenization cycles are completed prior to the washing of inclusion bodies. In total, seven process variations were compared and the resulting batches of inclusion bodies were solubilized and refolded to investigate the effect of early downstream processing on the product concentration in the final refolds. For the most efficient process variant, the recursive high pressure homogenization protocol led to the highest measured product concentration of 855 mg/L after refolding. Thus, the proposed recursive high pressure homogenization approach led to an estimated 18 % reduction of CO₂ footprint caused by urea, and increased the product yield per biomass from 5.17 g/kg to 7.84 g/kg compared to its linear wash counterpart — without introducing non-standard equipment or chemicals.

治疗性蛋白质通常在大肠杆菌的细胞内以聚集形式产生，称为包涵体，必须将其溶解并重新折叠以恢复生物活性产物。然而，工业上建立的分批稀释的折叠法仍然在低蛋白质浓度下进行，导致高用水量，设备占地面积大，可持续性差。提高再折叠产率从而使再折叠后的产物浓度更高的尚未开发的方法是优化细胞裂解和包涵体洗涤。在这项研究中，我们开发并评估了一种“递归高压均质”方法，其中洗涤步骤和额外的均质循环相结合，以提高溶解前包裹体的最终纯度。将细胞裂解程度、核酸释放程度和包涵体最终纯度与常规的“线性洗涤”进行比较，在常规的“线性洗涤”中，所有的均质循环都在包涵体洗涤之前完成。总共比较了7种工艺变化，并对所得到的包裹体进行了溶解和再折叠，以研究早期下游加工对最终再折叠时产品浓度的影响。对于最有效的工艺变体，递归高压均质方案导致折叠式后的最高测量产物浓度为855mg/L。因此，在不引入非标准设备或化学品的情况下，与线性洗涤方法相比，所提出的递归高压均质化方法估计可将尿素引起的二氧化碳足迹减少18%，并将每生物质的产品产量从5.17g/kg提高到7.84g/kg。

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引用次数: 0

CRISPR/anti-CRISPR genome editing in Clostridium beijerinckii 北京氏梭菌CRISPR/抗CRISPR基因组编辑。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-05 DOI: 10.1016/j.jbiotec.2025.11.002

Rémi Hocq , Gwladys Chartier, Nicolas Lopes Ferreira, François Wasels

The development of CRISPR technologies has revolutionized genome editing. However, in bacteria, CRISPR-based methods can be difficult to implement due to the cytotoxicity of CRISPR-associated proteins, which often impair or entirely prevent transformation. In this work, we combine inducible expression of classical CRISPR-Cas9 components with the anti-CRISPR protein AcrIIA4 from Listeria monocytogenes to tightly regulate Cas9 activity. Using this approach, we demonstrate efficient and iterative genome editing in the genetically recalcitrant Clostridium beijerinckii DSM 6423. While deletion of upp alone was not sufficient to render the strain sensitive to 5-fluorouracil, the additional deletion of a second gene involved in the uracil salvage pathway conferred resistance to the drug and validated our gene editing strategy. Collectively, our results show that CRISPR/anti-CRISPR systems can overcome a key limitation of CRISPR-based genome editing and may offer a broadly applicable strategy for engineering otherwise intractable bacterial species.

CRISPR技术的发展彻底改变了基因组编辑。然而，在细菌中，由于crispr相关蛋白的细胞毒性，基于crispr的方法可能难以实施，这通常会损害或完全阻止转化。在这项工作中，我们将经典CRISPR-Cas9组分的诱导表达与来自单核增生李斯特菌的抗crispr蛋白AcrIIA4结合起来，严格调节Cas9的活性。使用这种方法，我们证明了在遗传顽固性贝氏梭菌DSM 6423中有效和迭代的基因组编辑。虽然单独删除upp不足以使菌株对5-氟尿嘧啶敏感，但额外删除涉及尿嘧啶回收途径的第二个基因使其对药物产生耐药性，并验证了我们的基因编辑策略。总的来说，我们的研究结果表明，CRISPR/anti-CRISPR系统可以克服基于CRISPR的基因组编辑的一个关键限制，并可能为工程处理其他棘手的细菌物种提供广泛适用的策略。

引用次数: 0