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Harnessing Oxidative Stress to Obtain Natural Riboflavin Secreting Lactic Acid Bacteria for Use in Biofortification 利用氧化应激获取可分泌天然核黄素的乳酸菌用于生物强化
IF 6.1 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-14 DOI: 10.1021/acs.jafc.4c08881
Emmelie Joe Freudenberg Rasmussen, Norbert Acs, Peter Ruhdal Jensen, Christian Solem
Lactococcus lactis suffers from oxidative stress and riboflavin starvation at elevated temperatures due to dissolved oxygen, which can be relieved partially by exogenously supplied riboflavin. Here we explore whether this phenomenon can be harnessed to obtain riboflavin overproducing mutants. Using a riboflavin auxotrophic L. lactis strain as a riboflavin biosensor, we screened L. lactis cultures that had been exposed to temperature induced oxidative stress for up to one year. Riboflavin secreting mutants could readily be identified, some of which had arisen after just two weeks of exposure to oxidative stress. Whole genome sequencing revealed mutations in the riboswitch, which regulate riboflavin biosynthesis. Riboflavin secretion conferred a significant increase in tolerance to oxidative stress and enabled growth at high temperatures in the presence of dissolved oxygen. It was subsequently demonstrated that vigorous aeration at high temperature (37 °C) could prompt rapid emergence of riboflavin secreting mutants. The protective effect provided by riboflavin against oxidative stress may explain the natural occurrence of lactic acid bacteria (LAB) secreting riboflavin. By optimizing fermentation conditions and eliminating lactate formation, we achieved 64 mg/L riboflavin, the highest reported titer so far for LAB, which indicates great potential for use as a riboflavin fortification agent in food.
在高温条件下,乳酸乳球菌会因溶解氧而遭受氧化应激和核黄素饥饿。在此,我们探讨了能否利用这一现象获得核黄素过量产生的突变体。利用核黄素辅助营养型乳酸菌株作为核黄素生物传感器,我们筛选了暴露在温度诱导的氧化应激下长达一年的乳酸菌培养物。核黄素分泌突变体很容易被识别出来,其中一些突变体在暴露于氧化应激仅两周后就出现了。全基因组测序发现了核糖开关的突变,该开关能调节核黄素的生物合成。核黄素的分泌显著增强了对氧化应激的耐受性,并使其能够在有溶解氧的高温条件下生长。随后的研究证明,在高温(37 °C)下剧烈通气可促使核黄素分泌突变体迅速出现。核黄素对氧化应激的保护作用可以解释为什么天然乳酸菌(LAB)会分泌核黄素。通过优化发酵条件和消除乳酸盐的形成,我们获得了 64 mg/L 的核黄素,这是迄今为止报道的乳酸菌的最高滴度,这表明核黄素在食品中用作核黄素强化剂的巨大潜力。
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引用次数: 0
Comprehensive Overview of the Amide Linker Modification in the Succinate Dehydrogenase Inhibitors 琥珀酸脱氢酶抑制剂中酰胺连接体修饰的全面概述
IF 6.1 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-14 DOI: 10.1021/acs.jafc.4c05854
Ling-Qiang Zeng, Qi Chen, Ge Wei, Wei Chen, Xiao-Lei Zhu, Guang-Fu Yang
Succinate dehydrogenase inhibitors (SDHIs) have become one of the most important classes of agrochemical fungicides. According to the data from FRAC, the resistance risk for SDHIs had reached up to medium and even to high. In general, the chemical structure of SDHIs mainly contained three fragments: an acid core, a hydrophobic tail, and an amide linker, corresponding to three modification directions for each fragment. Among them, amide linker modification (ALM) has become a research hotspot for the design of novel SDHIs fungicides in recent years. We presented here a detailed review on the ALM strategy in the past decade, and some of them had entered the market. According to their chemical structures, ALM strategy were classified into four parts: (1) linked aliphatic chain between amide bond and hydrophobic tail, (2) introducing substituents to replacing hydrogen atom in the amide bond, (3) reverse extending the amide linker, and (4) changed with other bioisosteres. Moreover, the structure–activity relationship and the interaction mechanism of ALM-SDHI with SDH were discussed. This review aims to provide a global perspective on research and development of novel SDHIs, as well as suggestions for food safety management.
琥珀酸脱氢酶抑制剂(SDHIs)已成为最重要的农用杀菌剂之一。根据 FRAC 的数据,SDHIs 的抗药性风险已达到中度甚至高度。一般来说,SDHIs 的化学结构主要包括三个片段:酸核、疏水尾和酰胺连接体,每个片段对应三个修饰方向。其中,酰胺连接体修饰(ALM)已成为近年来设计新型SDHIs杀菌剂的研究热点。我们在此对近十年来的 ALM 策略进行了详细回顾,其中一些已进入市场。根据其化学结构,ALM策略可分为四个部分:(1)在酰胺键和疏水尾之间连接脂肪族链;(2)引入取代基取代酰胺键中的氢原子;(3)反向延伸酰胺连接体;(4)与其他生物异构体发生变化。此外,还讨论了 ALM-SDHI 与 SDH 的结构-活性关系和相互作用机制。本综述旨在为新型 SDHIs 的研究和开发提供一个全球视角,并为食品安全管理提供建议。
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引用次数: 0
Reactivity of the 2-Methylfuran Phase I Metabolite 3-Acetylacrolein Toward DNA. 2-Methylfuran Phase I Metabolite 3-Acetylacrolein 与 DNA 的反应性。
IF 5.7 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 Epub Date: 2024-11-04 DOI: 10.1021/acs.jafc.4c07280
Verena Schäfer, Simone Stegmüller, Hanna Becker, Elke Richling

2-Methylfuran (2-MF) is a well-known industrial chemical and also formed via thermal treatment of food. One main source of 2-MF in the human diet is coffee. 2-MF is known to form 3-acetylacrolein (AcA, 4-oxopent-2-enal) via cytochrome P 450 metabolism and further reacts with amino acids in vivo. Still the reactivity toward other biomolecules is rather scarce. Therefore, AcA was synthesized, and its reaction with 2'-deoxyadenosine (dA), 2'deoxyguanosine (dG), 2'deoxycytosine (dC), and 2'-deoxythymidine (dT) was tested. For this purpose, adduct formation was performed by acid hydrolysis of 2,5-dihydro-2,5-dimethoxy-2-methylfuran (DHDMMF) as well as pure AcA. The structures of these adducts were confirmed by UPLC-ESI+-MS/MS fragmentation patterns and 1H-/13CNMR spectra. Except for dT, which showed no reactivity, all adducts of AcA were characterized, which enabled the development of sensitive quantification methods via (U)HPLC-ESI±-MS/MS. Pure AcA was synthesized by oxidation of 2-MF using dimethyldioxirane (DMDO), and its behavior in aqueous medium was studied. Incubations of AcA and isolated DNA of primary rat hepatocytes (pRH) showed time- and dose-dependent formation of the identified DNA adducts dA-AcA, dG-AcA, or dC-AcA. In contrast, the DNA adducts dA-AcA, dG-AcA, or dC-AcA were not detected on a cellular level when pRH were incubated with 2-MF or AcA. This indicates an efficient detoxification or reaction with biomolecules in the cell, although the induction of other DNA damage, possibly also by other metabolites, cannot be ruled out in principle.

2-甲基呋喃(2-MF)是一种著名的工业化学品,也可通过热处理食物形成。人类饮食中 2-MF 的一个主要来源是咖啡。已知 2-MF 可通过细胞色素 P 450 代谢生成 3-乙酰基丙烯醛(AcA,4-氧代戊-2-烯醛),并进一步与体内的氨基酸发生反应。但是,这种物质对其他生物大分子的反应性还很少。因此,我们合成了 AcA,并测试了它与 2'-脱氧腺苷(dA)、2'-脱氧鸟苷(dG)、2'-脱氧胞嘧啶(dC)和 2'-脱氧胸苷(dT)的反应。为此,通过酸水解 2,5-二氢-2,5-二甲氧基-2-甲基呋喃(DHDMMF)和纯 AcA 来形成加合物。UPLC-ESI+-MS/MS 片段模式和 1H-/13CNMR 光谱证实了这些加合物的结构。除了 dT 没有反应外,所有 AcA 的加合物都得到了表征,从而可以通过 (U)HPLC-ESI±-MS/MS 开发出灵敏的定量方法。利用二甲基二氧环己烷(DMDO)氧化 2-MF 合成了纯 AcA,并研究了它在水介质中的行为。将 AcA 与原代大鼠肝细胞(pRH)分离出的 DNA 进行孵育,结果显示 DNA 加合物 dA-AcA、dG-AcA 或 dC-AcA 的形成具有时间和剂量依赖性。相反,当 pRH 与 2-MF 或 AcA 一起培养时,在细胞水平上检测不到 DNA 加合物 dA-AcA、dG-AcA 或 dC-AcA。这表明细胞内的解毒或与生物分子的反应是有效的,但原则上不能排除其他代谢物也可能诱发其他 DNA 损伤。
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引用次数: 0
Starch and Protein Characteristics of Chestnut Flours and Their Applications in Gluten-Free Products. 栗子粉的淀粉和蛋白质特性及其在无麸质产品中的应用
IF 5.7 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 Epub Date: 2024-11-04 DOI: 10.1021/acs.jafc.4c07201
Andrea Bresciani, Davide Russo, Mariasole Cervini, Chiara Magni, Gianluca Giuberti, Alessandra Marti

In the context of valorizing underutilized crops, this study investigated the starch and protein characteristics of chestnut varieties (Balestrera and Rossera) and their potential for use at 15% and 25% in reformulating gluten-free baked snacks and fresh pasta. Chestnut varieties differ in protein and starch content (Balestrera > Rossera), as well as in amylose, dietary fiber, and lipids (Balestrera < Rossera). Differences in starch and amylose content affected starch gelatinization (Balestrera < Rossera), pasting and retrogradation properties, and water absorption capacity at 90 °C (Balestrera > Rossera). No differences in water and oil absorption capacity and mixing properties were observed, neither in the protein profile nor in the nature of protein aggregates. Both varieties exhibited a good aptitude for producing gluten-free baked snacks and fresh pasta, even at enrichment levels of 25%. Overall, this study could provide valuable information for the development of gluten-free products using sustainable, underutilized crops.

在对未充分利用的作物进行价值评估的背景下,本研究调查了栗子品种(Balestrera 和 Rossera)的淀粉和蛋白质特性,以及它们在重新配制无麸质烘焙点心和新鲜面食中 15% 和 25% 的使用潜力。栗子品种在蛋白质和淀粉含量(Balestrera > Rossera)以及直链淀粉、膳食纤维和脂质(Balestrera < Rossera)方面存在差异。淀粉和直链淀粉含量的差异会影响淀粉糊化(巴勒斯特拉 < 罗塞拉)、糊化和逆变性以及 90 °C 时的吸水能力(巴勒斯特拉 > 罗塞拉)。在吸水率、吸油率和混合性能方面,无论是蛋白质还是蛋白质聚合体的性质都没有发现差异。这两个品种在生产无麸质烘焙点心和新鲜面食方面都表现出良好的适应性,即使富集度达到 25%。总之,这项研究可以为利用可持续的、未充分利用的作物开发无麸质产品提供有价值的信息。
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引用次数: 0
Lactobacillus rhamnosus 1.0320 Postbiotics Ameliorate Dextran Sodium Sulfate-Induced Colonic Inflammation and Oxidative Stress by Regulating the Intestinal Barrier and Gut Microbiota. 鼠李糖乳杆菌 1.0320 生后益生菌通过调节肠道屏障和肠道微生物群改善右旋糖酐硫酸钠诱发的结肠炎症和氧化应激。
IF 5.7 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 Epub Date: 2024-11-01 DOI: 10.1021/acs.jafc.4c06303
Jing Zhang, Xiaolei Duan, Xianhui Chen, Shanshan Qian, Jiage Ma, Zhanmei Jiang, Juncai Hou

Probiotics are increasingly being used as an adjunctive therapy for ulcerative colitis. However, some safety issues have been found in the clinical use of probiotics. Postbiotics have attracted much attention due to their storage stability, safety, and potential functions, but the dose required to exert a significant protective effect is unknown. Therefore, this study evaluated the potential mechanisms of different doses (200, 400, 600 mg/kg) of Lactobacillus rhamnosus 1.0320 postbiotics (1.0320P) in alleviating dextran sodium sulfate (DSS)-induced colitis. The study revealed that 1.0320P could mitigate DSS-induced colitis with signs of reductions in the disease activity index, amelioration of colon tissue damage, decreased secretion of proinflammatory cytokines, reduced oxidative stress levels, and lower bone marrow peroxidase activity. Furthermore, high dose of 1.0320P could upregulated the expression of key proteins in the Nrf2/ARE pathway (NQO1, Nrf2, and HO-1) and downregulated the expression of key proteins in the TLR4/NF-κB signaling pathway (TLR4, MyD88, and NF-κB p65). In addition, high dose of 1.0320P could upregulate the expression of tight junction (TJ) proteins including ZO-1, Occludin, and Claudin-1, contributing to the restoration of the intestinal mucosal barrier function. Additionally, 1.0320P was found to effectively correct imbalances in the intestinal microbiota and enhance the synthesis of short-chain fatty acids (SCFAs), thereby regulating homeostasis in the intestinal internal environment. Overall, our findings suggest that postbiotics could ameliorate colonic inflammation while being somewhat dose-dependent. This study provides new insights into postbiotics as a next-generation biotherapeutic agent for the treatment of ulcerative colitis and even other diseases.

益生菌越来越多地被用作溃疡性结肠炎的辅助疗法。然而,在益生菌的临床应用中也发现了一些安全性问题。益生菌后因其储存稳定性、安全性和潜在功能而备受关注,但产生显著保护作用所需的剂量尚不清楚。因此,本研究评估了不同剂量(200、400、600 毫克/千克)鼠李糖乳杆菌 1.0320 后益生菌(1.0320P)在缓解右旋糖酐硫酸钠(DSS)诱导的结肠炎方面的潜在机制。研究发现,1.0320P 可减轻右旋糖酐硫酸钠(DSS)诱导的结肠炎,表现为疾病活动指数降低、结肠组织损伤改善、促炎细胞因子分泌减少、氧化应激水平降低和骨髓过氧化物酶活性降低。此外,大剂量 1.0320P 还能上调 Nrf2/ARE 通路关键蛋白(NQO1、Nrf2 和 HO-1)的表达,下调 TLR4/NF-κB 信号通路关键蛋白(TLR4、MyD88 和 NF-κB p65)的表达。此外,高剂量的 1.0320P 还能上调包括 ZO-1、Occludin 和 Claudin-1 在内的紧密连接(TJ)蛋白的表达,从而有助于恢复肠粘膜屏障功能。此外,1.0320P 还能有效纠正肠道微生物群的失衡,促进短链脂肪酸 (SCFA) 的合成,从而调节肠道内部环境的平衡。总之,我们的研究结果表明,益生菌后可以改善结肠炎症,同时具有一定的剂量依赖性。这项研究为将益生菌作为治疗溃疡性结肠炎甚至其他疾病的下一代生物治疗药物提供了新的视角。
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引用次数: 0
Naphthoquinone Derivatives from the Endophytic Fungus Fusarium solani Induce Pancreatic Cancer Cells Apoptosis via Targeting EGFR-Mediated PI3K/Akt Pathway 内生真菌镰刀菌中的萘醌衍生物通过靶向表皮生长因子受体介导的 PI3K/Akt 通路诱导胰腺癌细胞凋亡
IF 6.1 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 DOI: 10.1021/acs.jafc.4c08652
Lin-Lin Gao, Xu-Tong Fang, Shu-Hui Zhao, Chen-Xiao Hui, Wei-Wei Huang, Yu-Qi Gao, Jin-Ming Gao
Seven new polyketide compounds, four naphthoquinone derivatives, neofusarubins A–D (1, 3, 5, and 18) and three graminin-like compounds, fusofuranones A–C (1921), together with 14 known naphthoquinone derivatives, were isolated from the solid fermentation of Fusarium solani, an endophytic fungus obtained from medicinal plant as tea, Camellia chrysantha. The structures of new compounds were elucidated based on chemical evidence and spectral data analysis (1D and 2D-NMR, HR-ESI-MS, ECD, SC-XRD). Among the isolated compounds tested, 2-acetonyl-3-methyl-5-hydroxy-7-methoxy-naphthazarin (11) exhibited the most potent inhibitory activity against pancreatic cancer in PANC-1, MiaPaCa-2, and BxPC-3 cells. Network pharmacology analysis revealed that compound 11 inhibited cell proliferation and promotion of apoptosis by targeting epidermal growth factor receptor (EGFR), which were confirmed by cellular thermal shift assay (CETSA), microscale thermophoresis (MST) and EGFR stably knockdown cells model assay, respectively. In addition, molecular mechanism studies in vitro showed that 11 could suppress the growth of pancreatic cancer cells by targeting EGFR and effectively inhibit downstream PI3K/Akt signaling pathway. Collectively, these findings provide a new EGFR targeting natural product for the treatment of pancreatic cancer.
从从药用植物茶花(Camellia chrysantha)中获得的内生真菌 Fusarium solani 的固体发酵中分离出了 7 个新的多酮化合物,其中包括 4 个萘醌衍生物--新柚木素 A-D(1、3、5 和 18)和 3 个类似柚木素的化合物--呋喃柚木素 A-C(19-21),以及 14 个已知的萘醌衍生物。根据化学证据和光谱数据分析(一维和二维 NMR、HR-ESI-MS、ECD、SC-XRD)阐明了新化合物的结构。在测试的分离化合物中,2-丙酮基-3-甲基-5-羟基-7-甲氧基萘甲萘醌(11)对 PANC-1、MiaPaCa-2 和 BxPC-3 细胞中的胰腺癌具有最强的抑制活性。网络药理学分析表明,化合物 11 通过靶向表皮生长因子受体(EGFR)抑制细胞增殖并促进细胞凋亡,这一点分别通过细胞热转移试验(CETSA)、微尺度热泳(MST)和 EGFR 稳定敲除细胞模型试验得到了证实。此外,体外分子机制研究表明,11 能通过靶向表皮生长因子受体抑制胰腺癌细胞的生长,并有效抑制下游 PI3K/Akt 信号通路。总之,这些发现为治疗胰腺癌提供了一种新的表皮生长因子受体靶向天然产物。
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引用次数: 0
Enhancing Manganese Peroxidase: Innovations in Genetic Modification, Screening Processes, and Sustainable Agricultural Applications 增强锰过氧化物酶:基因改造、筛选过程和可持续农业应用方面的创新
IF 6.1 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 DOI: 10.1021/acs.jafc.4c05878
Lu He, Mingchen Yan, Muhammad Naeem, Minghaonan Chen, Yong Chen, Zhong Ni, Huayou Chen
Manganese peroxidase (MnP), a vital extracellular enzyme for the degradation of lignin and other organic pollutants, has demonstrated immense potential for agricultural and environmental applications, including straw pretreatment, feed fermentation, mycotoxin degradation, and water treatment. However, current research remains in its exploratory phase, with naturally sourced MnP unable to meet industrial-scale demands and no mature commercial enzyme preparations available on the market. This comprehensive review innovatively constructs a framework for MnP research, probing into its molecular conformation and catalytic principles, while providing an overview of the advancements in high-throughput screening and In silco designing strategies. Specifically, this review focuses on the practical applications of MnP in sustainable agriculture, elaborating on its potential and challenges in straw resource utilization, efficient feed fermentation, mycotoxin control, and water quality improvement. Furthermore, this review summarizes the recent achievements in optimizing MnP activity through enzyme engineering techniques and discuss customized mutation strategies tailored to specific agricultural and environmental requirements, thereby laying a solid theoretical foundation and scientific basis for the industrial production and commercialization of MnP.
锰过氧化物酶(MnP)是一种降解木质素和其他有机污染物的重要细胞外酶,在农业和环境应用方面具有巨大潜力,包括秸秆预处理、饲料发酵、霉菌毒素降解和水处理。然而,目前的研究仍处于探索阶段,天然来源的 MnP 无法满足工业规模的需求,市场上也没有成熟的商业酶制剂。本综述创新性地构建了 MnP 研究框架,探究其分子构象和催化原理,同时概述了高通量筛选和 In silco 设计策略的进展。具体而言,本综述侧重于 MnP 在可持续农业中的实际应用,阐述了它在秸秆资源利用、高效饲料发酵、霉菌毒素控制和水质改善方面的潜力和挑战。此外,本综述还总结了通过酶工程技术优化 MnP 活性的最新成果,并讨论了针对特定农业和环境要求的定制突变策略,从而为 MnP 的工业化生产和商业化奠定坚实的理论基础和科学依据。
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引用次数: 0
Global Profiling of Protein Phosphorylation, Acetylation, and β-Hydroxybutyrylation in Nannochloropsis oceanica 拟南芥蛋白质磷酸化、乙酰化和 β-羟基丁酰化的全球剖析
IF 6.1 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 DOI: 10.1021/acs.jafc.4c05869
Lingyu Ouyang, Wuxin You, Ansgar Poetsch, Li Wei
Protein post-translational modifications (PTMs) regulate protein functions but remain poorly characterized in Nannochloropsis. This study examined three PTMs: lysine acetylation (Kac), lysine β-hydroxybutyrylation (Kbhb), and phosphorylation. Using LC-MS/MS, we identified 4571 Kac sites, 7812 Kbhb sites, and 6237 phosphorylation sites across 2455, 3109, and 2786 proteins, respectively. Subcellular localization analysis revealed significant overlaps between Kac and Kbhb proteins, primarily in the chloroplast, cytosol, and nucleus, while phosphorylated proteins were predominantly located in the nucleus and chloroplast. Motif analysis highlighted specific amino acid enrichments around modification sites, with several motifs conserved. Additionally, 529 proteins harbored all three PTMs, underscoring the potential regulatory interplay. Kac, Kbhb, and phosphorylated proteins were particularly abundant in glycolysis, the TCA cycle, carbon fixation, and lipid metabolism pathways, influencing energy production and lipid accumulation. Based on previous transcriptome data under nutrient-limited conditions, these frequently modified key enzymes appear to be vital components in the response to abiotic stress. The presence of histone modifications related to Kac and Kbhb might also point to the epigenetic regulation in gene expression and stress adaptation. This comprehensive PTM landscape in N. oceanica provides a foundation of valuable insights into future metabolic engineering and biotechnological applications.
蛋白质翻译后修饰(PTMs)调控蛋白质功能,但在拟南芥中仍鲜为人知。本研究考察了三种 PTM:赖氨酸乙酰化(Kac)、赖氨酸β-羟基丁酰化(Kbhb)和磷酸化。通过 LC-MS/MS,我们在 2455、3109 和 2786 个蛋白质中分别发现了 4571 个 Kac 位点、7812 个 Kbhb 位点和 6237 个磷酸化位点。亚细胞定位分析表明,Kac 和 Kbhb 蛋白之间有明显的重叠,主要分布在叶绿体、细胞质和细胞核中,而磷酸化蛋白则主要分布在细胞核和叶绿体中。基团分析强调了修饰位点周围特定氨基酸的富集,其中有几个基团是保守的。此外,有 529 个蛋白质同时含有这三种 PTM,突显了潜在的调控相互作用。Kac、Kbhb和磷酸化蛋白在糖酵解、TCA循环、碳固定和脂质代谢途径中特别丰富,影响着能量的产生和脂质的积累。根据以往营养限制条件下的转录组数据,这些频繁修饰的关键酶似乎是非生物胁迫响应的重要组成部分。与 Kac 和 Kbhb 相关的组蛋白修饰的存在也可能表明了基因表达和胁迫适应的表观遗传调控。N. oceanica 全面的 PTM 图谱为未来的代谢工程和生物技术应用提供了宝贵的基础。
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引用次数: 0
Excessive Supplement of l-Arginine Induces Myopia via Orchestrating the MEK-ERK-NO Signaling Pathway 过量补充精氨酸可通过协调 MEK-ERK-NO 信号通路诱发近视
IF 6.1 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 DOI: 10.1021/acs.jafc.4c09346
Bo Bao, Jinpeng Liu, Jizhao Xin, Xiangkun Zhou, Ruixue Zhang, Jiawen Hao, Zhaohui Yang, Miao Zhang, Zhongyu Ma, Xuewei Yin, Hongsheng Bi, Dadong Guo
Myopia can impact an individual’s quality of life and induce multiple complications. In this study, we randomly divided guinea pigs into the normal control (NC) group, the lens-induced myopia (LIM) group, the normal control plus l-arginine injection (l-Arg) group, and the normal control plus saline injection (NaCl) group. The animals in each group received the relevant treatments. The results showed that myopia was deeper in the LIM and l-Arg groups compared with the NC and NaCl groups. Our experimental results suggest that the excessive supplement of l-Arg will induce myopia onset and a MEK-ERK cascade that stimulates the downstream nNOS-MMP2 and NRF2-KEAP1 signaling pathways, which is accompanied by inflammatory responses, fibrosis changes, and oxidative stress. After receiving excessive arginine supplements, the experimental individuals showed myopic shifts and a similar pathological state as the LIM group. In conclusion, the supplement of excessive arginine may be a danger factor for myopia onset.
近视会影响个人的生活质量并诱发多种并发症。在这项研究中,我们将豚鼠随机分为正常对照(NC)组、透镜诱发近视(LIM)组、正常对照加注射精氨酸(l-Arg)组和正常对照加注射生理盐水(NaCl)组。各组动物均接受了相关治疗。结果表明,与 NC 组和 NaCl 组相比,LIM 组和 l-Arg 组的近视度数更深。我们的实验结果表明,过量补充 l-Arg 会诱导近视发生,并引起 MEK-ERK 级联反应,刺激下游的 nNOS-MMP2 和 NRF2-KEAP1 信号通路,同时伴随炎症反应、纤维化变化和氧化应激。补充过量精氨酸后,实验者出现近视偏移,病理状态与 LIM 组相似。总之,补充过量精氨酸可能是近视发病的危险因素。
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引用次数: 0
Efficient 7-Dehydrocholesterol Production by Multiple Metabolic Engineering of Diploid Saccharomyces cerevisiae. 通过二倍体酿酒酵母的多重代谢工程高效生产 7-去氢胆固醇
IF 5.7 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Pub Date : 2024-11-13 Epub Date: 2024-10-31 DOI: 10.1021/acs.jafc.4c07609
Ziqi Ye, Xianhao Xu, Yaokang Wu, Yanfeng Liu, Jianghua Li, Guocheng Du, Long Liu, Xueqin Lv

7-Dehydrocholesterol (7-DHC), a direct precursor of vitamin D3, has attracted increasing attention in microbial fermentation recently. In this study, 7-DHC biosynthesis in diploid Saccharomyces cerevisiae with robust ergosterol production was achieved by heterologous 24-dehydrocholesterol reductase expression, generating 44.1 mg/L 7-DHC, whereas the titer of ergosterol decreased by 40.5%. The ergosterol biosynthetic pathway was completely blocked by knocking out ERG6 and ERG5, affording a 4.2-fold increase in the 7-DHC titer. Subsequently, the facilitation of the mevalonate and the postsqualene pathways accompanied by elimination of transcriptional repressors enhanced 7-DHC synthesis, and the 7-DHC titer reached 738.5 mg/L in a shake flask. Further validation in a 50 L fermenter demonstrated that the 7-DHC titer reached 3.80 g/L within just 24 h, with productivity reaching 158.3 mg/L/h, setting a new benchmark as the highest reported to date. This study paves the way toward a large-scale and cost-effective manufacture of 7-DHC.

7-脱氢胆固醇(7-DHC)是维生素 D3 的直接前体,近年来在微生物发酵中越来越受到关注。本研究通过异源表达 24-脱氢胆固醇还原酶,在二倍体酿酒酵母中实现了 7-DHC 的生物合成,并产生了大量麦角固醇,产生了 44.1 mg/L 的 7-DHC,而麦角固醇的滴度则下降了 40.5%。通过敲除 ERG6 和 ERG5,麦角甾醇生物合成途径被完全阻断,从而使 7-DHC 滴度增加了 4.2 倍。随后,在消除转录抑制因子的同时,促进了甲羟戊酸和后葵醛途径,增强了 7-DHC 的合成,在振荡烧瓶中 7-DHC 滴度达到 738.5 毫克/升。在 50 升发酵罐中的进一步验证表明,7-DHC 滴度在短短 24 小时内就达到了 3.80 克/升,生产率达到 158.3 毫克/升/小时,为迄今为止报道的最高生产率设定了新的基准。这项研究为大规模、经济高效地生产 7-DHC 铺平了道路。
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Journal of Agricultural and Food Chemistry
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