Sophorolipids

槐糖脂
  • 文章类型: Journal Article
    表面活性剂是诱导表面张力降低的两亲性物质,washing,和乳化,并用于各种目的。最近,由可再生资源制造的具有高生物相容性的生物表面活性剂越来越受到关注。Sophorolipid(SL),一种来自Starmerellabombicola的生物表面活性剂,具有去污力和乳化性能,使其适用于家庭和个人护理应用。然而,SLs的毒理学数据有限。在这项研究中,我们使用SL进行了细胞毒性和皮肤刺激测试,揭示由SL引起的细胞毒性和皮肤刺激极低(logIC50=4.76mg/L),与个人护理产品中使用的溶剂和油相当。
    Surfactants are amphiphilic substances that induce surface tension reduction, washing, and emulsification and are used for various purposes. Recently, biosurfactants manufactured from renewable resources and with high biocompatibility have gained increasing attention. Sophorolipid (SL), a type of biosurfactant derived from Starmerella bombicola, possesses detergency and emulsification properties, making it suitable for household and personal care applications. However, there are limited toxicological data on SLs. In this study, we conducted cytotoxicity and skin-irritation tests using SLs, revealing that cytotoxicity and skin irritation induced by SLs were extremely low (logIC 50 = 4.76 mg/L) and equivalent to those of solvents and oils used in personal care products.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    由于与合成表面活性剂相比具有多种优势,因此不断发展的生物技术行业将大量注意力集中在生物表面活性剂上。这些好处包括全球公共卫生,环境可持续性,以及各部门对环保产品的需求不断增加。用生物表面活性剂替代可以减少高达8%的终生二氧化碳排放,避免释放到大气中的约150万吨温室气体。因此,对生物表面活性剂的需求急剧上升,占世界表面活性剂产量的10%(约1000万吨/年)。生物表面活性剂独特的两亲结构,由亲水和疏水成分组成,使这些分子在乳化中发挥基本功能,泡沫形成,去污力,和石油分散——所有这些都是各个部门高度重视的特征。今天,各种生物表面活性剂以商业规模生产用于食品,石油,和农业产业,以及制药和化妆品行业。我们提供了关于微生物生物表面活性剂的知识体系的全面分析,这些知识是在这项研究中随着时间的推移而获得的。我们还讨论了有效开发和使用生物表面活性剂需要克服的好处和障碍,以及它们现在和未来的工业用途。
    The growing biotechnology industry has focused a lot of attention on biosurfactants because of several advantages over synthetic surfactants. These benefits include worldwide public health, environmental sustainability, and the increasing demand from sectors for environmentally friendly products. Replacement with biosurfactants can reduce upto 8% lifetime CO2 emissions avoiding about 1.5 million tons of greenhouse gas released into the atmosphere. Therefore, the demand for biosurfactants has risen sharply occupying about 10% (∼10 million tons/year) of the world production of surfactants. Biosurfactants\' distinct amphipathic structure, which is made up of both hydrophilic and hydrophobic components, enables these molecules to perform essential functions in emulsification, foam formation, detergency, and oil dispersion-all of which are highly valued characteristic in a variety of sectors. Today, a variety of biosurfactants are manufactured on a commercial scale for use in the food, petroleum, and agricultural industries, as well as the pharmaceutical and cosmetic industries. We provide a thorough analysis of the body of knowledge on microbial biosurfactants that has been gained over time in this research. We also discuss the benefits and obstacles that need to be overcome for the effective development and use of biosurfactants, as well as their present and future industrial uses.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    不可避免的食物废物可能是工业生物技术的重要原料,而它们的增值可以为食品加工者提供附加值。然而,尽管它们丰富且成本低,食品加工商和消费者产生的这些食品废物的异质/混合性质导致碳和氮含量的高度可变性,以及特定的底物,在食物垃圾水解物中。这限制了它们在生物制品合成中的应用。这些废物通常用于厌氧消化和混合微生物培养,在通过纯微生物和单一微生物培养用于更高价值的生化生产中创造了显著的知识差距。为了直接调查这种知识差距,单个食品加工机产生的各种废物流被酶水解并表征,以及基材的可变性程度,碳,氮被量化。然后在实验室规模上阐明了使用细菌(Cupriavidusnecator)和古细菌(Haloferaxmediterranei)以及酵母(Starmerellabombicola)生产槐糖脂生物表面活性剂对水解产物变异性的活力和性能的影响。之后,在这项实验性概念验证研究中实施的策略,超越,深入讨论了改善工业规模的价值,解决了食物垃圾水解产物的高变异性,包括培养基标准化和高非选择性微生物生长相关产品的合成。所提供的见解将有益于旨在利用食物废物作为工业生物技术原料的未来努力。
    Unavoidable food wastes could be an important feedstock for industrial biotechnology, while their valorization could provide added value for the food processor. However, despite their abundance and low costs, the heterogeneous/mixed nature of these food wastes produced by food processors and consumers leads to a high degree of variability in carbon and nitrogen content, as well as specific substrates, in food waste hydrolysate. This has limited their use for bioproduct synthesis. These wastes are often instead used in anaerobic digestion and mixed microbial culture, creating a significant knowledge gap in their use for higher value biochemical production via pure and single microbial culture. To directly investigate this knowledge gap, various waste streams produced by a single food processor were enzymatically hydrolyzed and characterized, and the degree of variability with regard to substrates, carbon, and nitrogen was quantified. The impact of hydrolysate variability on the viability and performance of polyhydroxyalkanoates biopolymers production using bacteria (Cupriavidus necator) and archaea (Haloferax mediterranei) as well as sophorolipids biosurfactants production with the yeast (Starmerella bombicola) was then elucidated at laboratory-scale. After which, strategies implemented during this experimental proof-of-concept study, and beyond, for improved industrial-scale valorization which addresses the high variability of food waste hydrolysate were discussed in-depth, including media standardization and high non-selective microbial organisms growth-associated product synthesis. The insights provided would be beneficial for future endeavors aiming to utilize food wastes as feedstocks for industrial biotechnology.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    发酵过程是动态变化的,通过对环境参数的实时监测可以掌握代谢状态。在这项研究中,开发了基于非接触式近红外(NIR)光谱技术的基板和产品检测实时在线监测实验平台。乳酸发酵过程监测的预测模型,基于偏最小二乘回归和内部交叉验证方法,建立了槐糖脂(SLs)和葡萄糖酸钠(SG)。通过发酵验证,NIR模型对复杂发酵环境的准确性和精度,不同的流变特性(均匀系统和多相不均匀系统)和不同的参数类型(底物,产品和营养素)具有良好的适用性,R2大于0.98,呈现良好的线性关系。预测的均方根误差表明该模型具有较高的可信度。通过NIR模型控制SG发酵中合适的葡萄糖浓度以及葡萄糖和油的浓度,SG和SL的滴度分别提高到11.8%和26.8%,分别。尽管NIR光谱仪的高成本是其在工业规模中广泛应用的关键问题。该工作为近红外光谱在复杂发酵体系中的应用提供了基础。
    The fermentation process is dynamically changing, and the metabolic status can be grasped through real-time monitoring of environmental parameters. In this study, a real-time and on-line monitoring experiment platform for substrates and products detection was developed based on non-contact type near-infrared (NIR) spectroscopy technology. The prediction models for monitoring the fermentation process of lactic acid, sophorolipids (SLs) and sodium gluconate (SG) were established based on partial least-squares regression and internal cross-validation methods. Through fermentation verification, the accuracy and precision of the NIR model for the complex fermentation environments, different rheological properties (uniform system and multi-phase inhomogeneous system) and different parameter types (substrate, product and nutrients) have good applicability, and R2 was greater than 0.98, exhibiting a good linear relationship. The root mean square error of prediction shows that the model has high credibility. Through the control of appropriate glucose concentration in SG fermentation as well as glucose and oil concentrations SLs fermentation by NIR model, the titers of SG and SLs were increased to 11.8% and 26.8%, respectively. Although high cost of NIR spectrometer is a key issue for its wide application in an industrial scale. This work provides a basis for the application of NIR spectroscopy in complex fermentation systems.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    在这项研究中,研究了生物表面活性剂槐糖脂(SLs)对米曲霉根瘤菌脂肪酶(RML)发酵的影响。在初始培养基中外源添加0.3%(w/v)SLs,RML活性达到430.0U/mL,与对照组相比增加了25.0%。随后,进一步探讨了米曲霉对添加SLs的生理代谢反应。结果表明,尽管SL对RML分泌几乎没有影响,它会影响细胞的形态。在发酵后期,中间颗粒的比例,随着SL的添加,通常被认为是一种充满活力和稳定的酶生产状态。同时,发酵液的粘度降低,这促进了氧气转移的增加,从而提高RML产量。最后,可以发现,添加SLs可以显着增加前体氨基酸的含量,尤其是那些排名第一和第二的RML组成,能促进RML的合成。
    In this study, the effect of biosurfactant sophorolipids (SLs) on Rhizomucor miehei lipase (RML) fermentation by Aspergillus oryzae was investigated. With the exogenous addition of 0.3% (w/v) SLs in the initial medium, the RML activity reached 430.0 U/mL, an increase of 25.0% compared to the control group. Subsequently, the physiological metabolic responses of A. oryzae to the addition of SLs were further explored. The results showed that though SLs had almost no effect on the RML secretion, it would affect the morphology of the cells. During the late phase of the fermentation, the proportion of middle pellets, which was generally considered as an energetic and stable state for enzyme production was increased with the addition of SLs. Simultaneously, the viscosity of fermentation broth was reduced, which facilitated the increase of oxygen transfer, thereby improving the RML production. Finally, it could be found that the addition of SLs significantly increased the contents of precursor amino acids, especially for those rank first and second of the RML composition, and it could promote the synthesis of RML.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    用生物分子替代农用化学品对于减轻土壤污染和其核心微生物群的失活至关重要。在此背景下,这项研究旨在研究诸如哈茨木霉CCT2160(BF-Th)之类的生物防治剂与源自巴西本地酵母StarmorellabombicolaUFMG-CM-Y6419的生物表面活性剂(BS)之间的相互作用。此后,测试了它们在水稻种子萌发中的潜力。两种生物产品均在现场生产,并通过BS的HPLC-MS和GC-MS以及BF-Th的SDS-PAGE凝胶根据其化学组成进行表征。BS被证实是槐糖脂(SL),其是充分研究的具有抗微生物活性的化合物。通过在固体和深层发酵中添加0.1至2g/L的SL来培养真菌,以检查其生物相容性。在固态发酵中,补充SL可提高孢子产量,赋予两种生物产品的协同作用。对于发芽测定,由SL组成的生物制剂,BF-Th和组合(SLT)应用于O.sativaL种子的萌发,在形态方面,例如根和芽大小以及侧根的存在,提高了30%。假设SL能够在生长的早期调节植物激素的表达,例如生长素和赤霉素。指向他们新颖的植物生长刺激特性。因此,这项研究指出了由生物表面活性剂和活性内生真菌孢子组成的混合生物制剂的潜力,以增强植物适应性和可能的疾病控制。
    The replacement of agrochemicals by biomolecules is imperative to mitigate soil contamination and inactivation of its core microbiota. Within this context, this study aimed at the interaction between a biological control agent such as Trichoderma harzianum CCT 2160 (BF-Th) and the biosurfactants (BSs) derived from the native Brazilian yeast Starmerella bombicola UFMG-CM-Y6419. Thereafter, their potential in germination of Oryza sativa L. seeds was tested. Both bioproducts were produced on site and characterized according to their chemical composition by HPLC-MS and GC-MS for BSs and SDS-PAGE gel for BF-Th. The BSs were confirmed to be sophorolipids (SLs) which is a well-studied compound with antimicrobial activity. The biocompatibility was examined by cultivating the fungus with SLs supplementation ranging from 0.1 to 2 g/L in solid and submerged fermentation. In solid state fermentation the supplementation of SLs enhanced spore production, conferring the synergy of both bioproducts. For the germination assays, bioformulations composed of SLs, BF-Th and combined (SLT) were applied in the germination of O. sativa L seeds achieving an improvement of up to 30% in morphological aspects such as root and shoot size as well as the presence of lateral roots. It was hypothesized that SLs were able to regulate phytohormones expression such as auxins and gibberellins during early stage of growth, pointing to their novel plant-growth stimulating properties. Thus, this study has pointed to the potential of hybrid bioformulations composed of biosurfactants and active endophytic fungal spores in order to augment the plant fitness and possibly the control of diseases.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    槐糖脂,糖脂生物表面活性剂来自微生物,如Starmerellabombicola,具有独特的表面活性和生物活性,持有化妆品的潜在应用,药物和生物修复。然而,野生型槐糖脂有限的结构变异性限制了它们的性质和应用。为了解决这个问题,代谢工程的努力已经允许创建一个分子组合。在这项研究中,我们通过化学修饰微生物产生的槐糖苷更进一步,由工程化的S.bombicola生产。合成了24种新的苦参苷衍生物,包括在氮原子上具有不同烷基链长度(乙基至十八烷基)的槐糖胺及其相应的季铵盐。此外,将六种不同的微生物产生的糖脂生物表面活性剂氢化以获得完全饱和的脂质尾巴。这些衍生物,以及微生物产生的糖脂和三种基准生物表面活性剂(双鼠李糖脂,烷基多葡糖苷,椰油酰胺丙基甜菜碱),对细菌(枯草芽孢杆菌,金黄色葡萄球菌,单核细胞增生李斯特菌,大肠杆菌,铜绿假单胞菌)和酵母(白色念珠菌)。结果表明,微生物产生的糖脂,比如Bolasophosides,酸性槐糖脂和酸性葡萄糖脂表现出对测试生物体的选择性抗微生物活性。相反,乳酸槐糖脂,槐苷胺和季铵盐显示出广泛的抗菌活性。正辛基,N-十二烷基和N-十八烷基衍生物表现出最低的最低抑制浓度,范围从0.014到20.0mgmL-1。这项研究证明了周到的生物技术和目标化学的潜在协同作用,可以精确地定制糖脂生物表面活性剂,以满足各种应用的特定要求。
    Sophorolipids, glycolipid biosurfactants derived from microorganisms such as Starmerella bombicola, possess distinctive surface-active and bioactive properties, holding potential applications in cosmetics, pharmaceuticals and bioremediation. However, the limited structural variability in wild-type sophorolipids restricts their properties and applications. To address this, metabolic engineering efforts have allowed to create a portfolio of molecules. In this study, we went one step further by chemically modifying microbially produced sophorosides, produced by an engineered S. bombicola. Twenty-four new sophoroside derivatives were synthesized, including sophoroside amines with varying alkyl chain lengths (ethyl to octadecyl) on the nitrogen atom and their corresponding quaternary ammonium salts. Additionally, six different microbially produced glycolipid biosurfactants were hydrogenated to achieve fully saturated lipid tails. These derivatives, along with microbially produced glycolipids and three benchmark biosurfactants (di-rhamnolipids, alkyl polyglucosides, cocamidopropyl betaine), were assessed for antimicrobial activity against bacteria (Bacillus subtilis, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa) and yeast (Candida albicans). Results indicated that microbially produced glycolipids, such as bola sophorosides, acidic sophorolipids and acidic glucolipids exhibit selective antimicrobial activity against the test organisms. Conversely, lactonic sophorolipids, sophoroside amines and quaternary ammonium salts display a broad antimicrobial activity. N-octyl, N-dodecyl and N-octadecyl derivatives exhibit the lowest minimal inhibitory concentrations, ranging from 0.014 to 20.0 mg mL-1. This study demonstrates the potential synergy of thoughtful biotechnology and targeted chemistry to precisely tailor glycolipid biosurfactants to meet specific requirements across applications.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    皮肤病,包括寻常痤疮,特应性皮炎,还有酒渣鼻,以生物膜的存在为特征,它们是微生物群落。生物膜的机械稳定性归因于微生物分泌的其成分之一-多糖。槐糖脂是具有生物膜破坏和去除能力的生物表面活性剂,有望成为化妆品中经典石化基表面活性剂的替代品。在这项研究中,我们研究了槐糖脂对β-葡聚糖的影响,如分散状态,相互作用机制,和构型改变为水溶液中生物膜的模型多糖。动态光散射测量表明,槐糖脂干扰β-葡聚糖在水溶液中的聚集。相比之下,十二烷基硫酸钠(SDS),用作典型的表面活性剂参考,促进β-葡聚糖的聚集。使用表面张力测量和等温滴定量热法(ITC)研究了槐糖脂与β-葡聚糖之间的相互作用。在存在β-葡聚糖的情况下,表面张力仅在槐糖脂的临界胶束浓度(CMC)附近增加。这表明相互作用发生在溶液中而不是在气液界面处。此外,ITC的结果表明疏水相互作用参与了这种相互作用。此外,旋光测量的结果表明,槐糖脂没有展开β-葡聚糖的三螺旋结构。当槐糖脂通过疏水相互作用与β-葡聚糖相互作用时,预期β-葡聚糖分散体会引起空间位阻和静电排斥,这归因于庞大的糖部分和羧基官能团的槐糖脂的独特分子结构。这些结果证明了槐糖脂对β-葡聚糖的独特性能,并提供了关于槐糖脂作为良好抗生物膜功效的更多见解。
    Skin disorders, including acne vulgaris, atopic dermatitis, and rosacea, are characterized by the presence of biofilms, which are communities of microorganisms. The mechanical stability of biofilms is attributed to one of their constituents-polysaccharides-which are secreted by microorganisms. Sophorolipids are biosurfactants with biofilm disruption and removal abilities and are expected to become alternatives for classical petrochemical-based surfactants in cosmetics. In this study, we investigated the influence of sophorolipids on β-glucan such as dispersion status, interaction mechanism, and configuration change as a model polysaccharide of biofilm in aqueous solution. Dynamic light scattering measurements showed that sophorolipids interfere with the aggregation of β- glucan in aqueous solutions. In contrast, sodium dodecyl sulfate (SDS), which is used as a typical surfactant reference, promotes the aggregation of β-glucan. The interaction between sophorolipids and β-glucan were investigated using surface tension measurements and isothermal titration calorimetry (ITC). Surface tension increased only near critical micelle concentration (CMC) region of sophorolipids in the presence of β-glucan. This suggests that the interaction occurred in the solution rather than at the air-liquid interface. Moreover, the results of ITC indicate that hydrophobic interactions were involved in this interaction. In addition, the results of optical rotation measurements indicate that sophorolipids did not unfold the triple helical structure of β-glucan. β-glucan dispersion was expected to be caused steric hindrance and electrostatic repulsion when sophorolipids interacted with β-glucan via hydrophobic interactions owing to the unique molecular structure of sophorolipids attributed by a bulky sugar moiety and a carboxyl functional group. These results demonstrated unique performances of sophorolipids on β-glucan and provided more insights on the efficacy of sophorolipids as good anti-biofilms.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    背景:阿霉素(DOX)是一种众所周知的化学治疗剂,由于多器官损伤而引起严重的不良反应,包括心脏毒性,肾毒性,神经毒性,和肝毒性。DOX诱导的器官毒性的机制可能归因于氧化应激(OS)和,因此,炎症信号通路的激活,凋亡,和阻断自噬。作为糖脂类型的生物表面活性剂,是天然产品,具有独特的性能和广泛的应用归因于其抗氧化和抗炎特性。
    目的:从香蕉果皮上生长的酿酒酵母生产低成本SL,并研究其对DOX诱导的肝毒性的可能保护作用。
    方法:对酵母进行局部分离和分子鉴定,然后用FTIR对产生的SL进行了表征,1HNMR和LC-MS/MS光谱。后期,将32只雄性Wistar大鼠随机分为4组;对照组(口服盐水),SLs(200mg/kg,p.o),DOX(10mg/kg;i.p.),和SL+DOX(200mg/kgp.o.,10mg/kg;i.p.分别)。肝功能测试(LFTs),氧化应激,炎症,细胞凋亡以及自噬标志物进行了研究。
    结果:SL的产量为49.04%,用SL处理可改善LFT,增强Nrf2和抑制NF-κB,IL-6,IL-1β,p38、caspase一3和Bax/Bcl2比值除增进自噬外,与DOX组比拟。
    结论:我们的研究结果揭示了SLs对抗DOX诱导的大鼠肝毒性的新的有希望的保护作用。
    BACKGROUND: Doxorubicin (DOX) is a well-known chemotherapeutic agent which causes serious adverse effects due to multiple organ damage, including cardiotoxicity, nephrotoxicity, neurotoxicity, and hepatotoxicity. The mechanism of DOX-induced organ toxicity might be attributed to oxidative stress (OS) and, consequently, activation of inflammatory signaling pathways, apoptosis, and blockage of autophagy. Sophorolipids (SLs) as a glycolipid type of biosurfactants, are natural products that have unique properties and a wide range of applications attributed to their antioxidant and anti-inflammatory properties.
    OBJECTIVE: Production of low-cost SLs from Saccharomyces cerevisiae grown on banana peels and investigating their possible protective effects against DOX-induced hepatotoxicity.
    METHODS: The yeast was locally isolated and molecularly identified, then the yielded SLs were characterized by FTIR, 1H NMR and LC-MS/MS spectra. Posteriorly, thirty-two male Wistar rats were randomly divided into four groups; control (oral saline), SLs (200 mg/kg, p.o), DOX (10 mg/kg; i.p.), and SL + DOX (200 mg/kg p.o.,10 mg/kg; i.p., respectively). Liver function tests (LFTs), oxidative stress, inflammatory, apoptosis as well as autophagy markers were investigated.
    RESULTS: SLs were produced with a yield of 49.04% and treatment with SLs improved LFTs, enhanced Nrf2 and suppressed NF-κB, IL-6, IL-1β, p38, caspase 3 and Bax/Bcl2 ratio in addition to promotion of autophagy when compared to DOX group.
    CONCLUSIONS: Our results revealed a novel promising protective effect of SLs against DOX-induced hepatotoxicity in rats.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    槐糖脂(SL)代表了非常有前途的生物表面活性剂。然而,由于涉及大量成本,其广泛的生产和应用遇到了障碍。这里,阐明了发酵过程与原位分离生产模式相结合的智能精确调节策略,实现具有成本效益的SLs生产。首先,构建了一个机制辅助的数据驱动模型,用于“按需喂养细胞”。此外,开发了一种基于细胞代谢能力需求的逐步氧气供应调节策略,完成了“细胞的按需氧气供应”,优化能耗控制。最后,通过将半连续发酵模式与SLs生产的原位分离技术相结合,实现了系统的方法。此策略提高了SL的生产率和产量,达到2.30g/L/h和0.57g/g,分别。与补料分批发酵相比,这些值代表40.2%和18.7%的增加。此外,分离后的粗SLs浓度达到793.12g/L,促进下游分离和纯化过程。
    Sophorolipids (SLs) represent highly promising biosurfactants. However, its widespread production and application encounter obstacles due to the significant costs involved. Here, an intelligent and precise regulation strategy was elucidated for the fermentation process coupled with in-situ separation production mode, to achieve cost-effective SLs production. Firstly, a mechanism-assisted data-driven model was constructed for \"on-demand feeding of cells\". Moreover, a strategy of step-wise oxygen supply regulation based on the demand for cell metabolic capacity was developed, which accomplished \"on-demand oxygen supply of cells\", to optimize the control of energy consumption. Finally, a systematic approach was implemented by integrating a semi-continuous fermentation mode with in-situ separation technology for SLs production. This strategy enhanced SLs productivity and yield, reaching 2.30 g/L/h and 0.57 g/g, respectively. These values represented a 40.2% and 18.7% increase compared to fed-batch fermentation. Moreover, the concentration of crude SLs after separation reached 793.12 g/L, facilitating downstream separation and purification processes.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

公众号