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Riboflavin overproduction on lignocellulose hydrolysate by the engineered yeast Candida famata. 工程酵母法氏念珠菌在木质纤维素水解物上过量产生核黄素。
IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foae020
Ljubov S Dzanaeva, Dominik Wojdyła, Dariya V Fedorovych, Justyna Ruchala, Kostyantyn V Dmytruk, Andriy A Sibirny

Lignocellulose (dry plant biomass) is an abundant cheap inedible residue of agriculture and wood industry with great potential as a feedstock for biotechnological processes. Lignocellulosic substrates can serve as valuable resources in fermentation processes, allowing the production of a wide array of chemicals, fuels, and food additives. The main obstacle for cost-effective conversion of lignocellulosic hydrolysates to target products is poor metabolism of the major pentoses, xylose and L-arabinose, which are the second and third most abundant sugars of lignocellulose after glucose. We study the oversynthesis of riboflavin in the flavinogenic yeast Candida famata and found that all major lignocellulosic sugars, including xylose and L-arabinose, support robust growth and riboflavin synthesis in the available strains of C. famata. To further increase riboflavin production from xylose and lignocellulose hydrolysate, genes XYL1 and XYL2 coding for xylose reductase and xylitol dehydrogenase were overexpressed. The resulting strains exhibited increased riboflavin production in both shake flasks and bioreactors using diluted hydrolysate, reaching 1.5 g L-1.

木质纤维素(干植物生物质)是农业和木材工业中大量廉价的不可食用残留物,具有作为生物技术工艺原料的巨大潜力。木质纤维素基质可作为发酵过程中的宝贵资源,生产出多种化学品、燃料和食品添加剂。将木质纤维素水解物转化为目标产品的成本效益的主要障碍是木质纤维素中仅次于葡萄糖的第二和第三大糖--主要戊糖木糖和 L-阿拉伯糖的代谢不良。我们研究了产黄酵母家庭念珠菌核黄素的过度合成,发现所有主要木质纤维素糖类,包括木糖和 L-阿拉伯糖,都能支持家庭念珠菌现有菌株的旺盛生长和核黄素合成。为了进一步提高木糖和木质纤维素水解物的核黄素产量,过量表达了编码木糖还原酶和木糖醇脱氢酶的基因 XYL1 和 XYL2。由此产生的菌株在使用稀释水解物的摇瓶和生物反应器中都表现出核黄素产量增加,达到 1.5 g L-1。
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引用次数: 0
Correction to: Yca1 metacaspase: diverse functions determine how yeast live and let die. 更正:Yca1元天冬酶:多种功能决定酵母如何生存和死亡
IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foae019
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引用次数: 0
Heterologous pulcherrimin production in Saccharomyces cerevisiae confers inhibitory activity on Botrytis conidiation. 在酿酒酵母中异源生产 pulcherrimin 可对肉毒菌分生孢子产生抑制作用。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foad053
Florian M Freimoser, Marina Mahler, Mark McCullough, Alexander O Brachmann, Lukas Nägeli, Maja Hilber-Bodmer, Jörn Piel, Stefan A Hoffmann, Yizhi Cai

Pulcherrimin is an iron (III) chelate of pulcherriminic acid that plays a role in antagonistic microbial interactions, iron metabolism, and stress responses. Some bacteria and yeasts produce pulcherriminic acid, but so far, pulcherrimin could not be produced in Saccharomyces cerevisiae. Here, multiple integrations of the Metschnikowia pulcherrima PUL1 and PUL2 genes in the S. cerevisiae genome resulted in red colonies, which indicated pulcherrimin formation. The coloration correlated positively and significantly with the number of PUL1 and PUL2 genes. The presence of pulcherriminic acid was confirmed by mass spectrometry. In vitro competition assays with the plant pathogenic fungus Botrytis caroliana revealed inhibitory activity on conidiation by an engineered, strong pulcherrimin-producing S. cerevisiae strain. We demonstrate that the PUL1 and PUL2 genes from M. pulcherrima, in multiple copies, are sufficient to transfer pulcherrimin production to S. cerevisiae and represent the starting point for engineering and optimizing this biosynthetic pathway in the future.

Pulcherrimin 是 Pulcherriminic 酸的铁(III)螯合物,在拮抗微生物相互作用、铁代谢和应激反应中发挥作用。一些细菌和酵母会产生 pulcherriminic 酸,但迄今为止,酿酒酵母还不能产生 pulcherrimin。在这里,将 Metschnikowia pulcherrima PUL1 和 PUL2 基因多次整合到酿酒酵母基因组中,结果产生了红色菌落,这表明形成了 pulcherrimin。颜色与 PUL1 和 PUL2 基因的数量呈显著正相关。质谱分析证实了pulcherriminic酸的存在。与植物病原真菌 Botrytis caroliana 的体外竞争试验表明,经过改造的强产pulcherrimin的 S. cerevisiae 菌株对分生孢子有抑制作用。我们证明,多拷贝的浆果霉 PUL1 和 PUL2 基因足以将浆果芸香素的生产转移到 S. cerevisiae 上,这也是未来工程化和优化这一生物合成途径的起点。
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引用次数: 0
Data integration strategies for whole-cell modeling. 全细胞建模的数据整合策略。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foae011
Katja Tummler, Edda Klipp

Data makes the world go round-and high quality data is a prerequisite for precise models, especially for whole-cell models (WCM). Data for WCM must be reusable, contain information about the exact experimental background, and should-in its entirety-cover all relevant processes in the cell. Here, we review basic requirements to data for WCM and strategies how to combine them. As a species-specific resource, we introduce the Yeast Cell Model Data Base (YCMDB) to illustrate requirements and solutions. We discuss recent standards for data as well as for computational models including the modeling process as data to be reported. We outline strategies for constructions of WCM despite their inherent complexity.

数据让世界运转--高质量的数据是建立精确模型的先决条件,尤其是全细胞模型(WCM)。全细胞模型的数据必须可重复使用,包含准确的实验背景信息,并应全面覆盖细胞中的所有相关过程。在此,我们回顾了 WCM 数据的基本要求以及如何将它们结合起来的策略。作为特定物种的资源,我们介绍了酵母细胞模型数据库(YCMDB),以说明要求和解决方案。我们讨论了数据和计算模型的最新标准,包括作为数据报告的建模过程。尽管 WCM 本身很复杂,但我们还是概述了构建 WCM 的策略。
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引用次数: 0
Exploring yeast biodiversity and process conditions for optimizing ethylene glycol conversion into glycolic acid. 探索酵母生物多样性和工艺条件,优化乙二醇转化为乙醇酸的过程。
IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foae024
Vittorio Giorgio Senatore, Riccardo Milanesi, Fiorella Masotti, Letizia Maestroni, Stefania Pagliari, Ciro Cannavacciuolo, Luca Campone, Immacolata Serra, Paola Branduardi

Plastics have become an indispensable material in many fields of human activities, with production increasing every year; however, most of the plastic waste is still incinerated or landfilled, and only 10% of the new plastic is recycled even once. Among all plastics, polyethylene terephthalate (PET) is the most produced polyester worldwide; ethylene glycol (EG) is one of the two monomers released by the biorecycling of PET. While most research focuses on bacterial EG metabolism, this work reports the ability of Saccharomyces cerevisiae and nine other common laboratory yeast species not only to consume EG, but also to produce glycolic acid (GA) as the main by-product. A two-step bioconversion of EG to GA by S. cerevisiae was optimized by a design of experiment approach, obtaining 4.51 ± 0.12 g l-1 of GA with a conversion of 94.25 ± 1.74% from 6.21 ± 0.04 g l-1 EG. To improve the titer, screening of yeast biodiversity identified Scheffersomyces stipitis as the best GA producer, obtaining 23.79 ± 1.19 g l-1 of GA (yield 76.68%) in bioreactor fermentation, with a single-step bioprocess. Our findings contribute in laying the ground for EG upcycling strategies with yeasts.

塑料已成为人类活动中许多领域不可或缺的材料,产量逐年增加;然而,大部分塑料垃圾仍被焚烧或填埋,只有 10%的新塑料被回收利用过一次。在所有塑料中,聚对苯二甲酸乙二醇酯(PET)是全球产量最大的聚酯;乙二醇(EG)是 PET 生物再循环过程中释放的两种单体之一。大多数研究都集中在细菌的乙二醇新陈代谢上,而这项工作报告了酿酒酵母和其他九种常见的实验室酵母不仅能消耗乙二醇,还能产生乙醇酸(GA)作为主要副产品。通过实验设计方法,对酿酒酵母将 EG 转化为 GA 的两步生物转化进行了优化,从 6.21 ± 0.04 g L-1 的 EG 中获得了 4.51 ± 0.12 g L-1 的 GA,转化率为 94.25 ± 1.74%。为了提高滴度,对酵母生物多样性进行了筛选,确定了 Scheffersomyces stipitis 是 GA 的最佳生产者,它在生物反应器发酵中获得了 23.79 ± 1.19 g L-1 的 GA(产量为 76.68%),采用的是单步生物工艺。我们的研究结果为利用酵母菌实现 EG 循环利用战略奠定了基础。
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引用次数: 0
Sphingolipid diversity in Candida auris: unraveling interclade and drug resistance fingerprints. 白色念珠菌的鞘脂多样性:揭示支链间和耐药性指纹。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foae008
Basharat Ali, Mohit Kumar, Praveen Kumar, Anshu Chauhan, Sana Akhtar Usmani, Shivaprakash M Rudramurthy, Jacques F Meis, Arunaloke Chakrabarti, Ashutosh Singh, Naseem A Gaur, Alok K Mondal, Rajendra Prasad

In this study, we explored the sphingolipid (SL) landscape in Candida auris, which plays pivotal roles in fungal biology and drug susceptibility. The composition of SLs exhibited substantial variations at both the SL class and molecular species levels among clade isolates. Utilizing principal component analysis, we successfully differentiated the five clades based on their SL class composition. While phytoceramide (PCer) was uniformly the most abundant SL class in all the isolates, other classes showed significant variations. These variations were not limited to SL class level only as the proportion of different molecular species containing variable number of carbons in fatty acid chains also differed between the isolates. Also a comparative analysis revealed abundance of ceramides and glucosylceramides in fluconazole susceptible isolates. Furthermore, by comparing drug-resistant and susceptible isolates within clade IV, we uncovered significant intraclade differences in key SL classes such as high PCer and low long chain base (LCB) content in resistant strains, underscoring the impact of SL heterogeneity on drug resistance development in C. auris. These findings shed light on the multifaceted interplay between genomic diversity, SLs, and drug resistance in this emerging fungal pathogen.

在这项研究中,我们探索了白色念珠菌中的鞘磷脂(SL)结构,它在真菌生物学和药物敏感性方面发挥着关键作用。在各支系分离物中,鞘脂的组成在鞘脂类别和分子物种水平上都表现出很大的差异。利用主成分分析法,我们成功地根据 SL 种类的组成将五个支系区分开来。虽然植酸酰胺(PCer)在所有分离物中都是含量最高的 SL 类,但其他类别也有显著差异。这些变化不仅限于 SL 类,因为不同分离物中脂肪酸链中含有不同碳数的不同分子种类的比例也不相同。比较分析还发现,对氟康唑敏感的分离物中含有大量的神经酰胺和葡萄糖基甘油酰胺。此外,通过比较第四支系中的耐药分离物和易感分离物,我们发现耐药菌株的关键 SL 类(如 PCer 含量高、长链基(LCB)含量低)在支系内存在显著差异,这凸显了 SL 异质性对蛔虫耐药性发展的影响。这些发现揭示了这种新兴真菌病原体的基因组多样性、SL 和耐药性之间的多方面相互作用。
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引用次数: 0
Application of unimodal probability distribution models for morphological phenotyping of budding yeast. 单模态概率分布模型在芽殖酵母形态表型中的应用
IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-01-09 DOI: 10.1093/femsyr/foad056
Yoshikazu Ohya, Farzan Ghanegolmohammadi, Kaori Itto-Nakama

Morphological phenotyping of the budding yeast Saccharomyces cerevisiae has helped to greatly clarify the functions of genes and increase our understanding of cellular functional networks. It is necessary to understand cell morphology and perform quantitative morphological analysis (QMA) but assigning precise values to morphological phenotypes has been challenging. We recently developed the Unimodal Morphological Data image analysis pipeline for this purpose. All true values can be estimated theoretically by applying an appropriate probability distribution if the distribution of experimental values follows a unimodal pattern. This reliable pipeline allows several downstream analyses, including detection of subtle morphological differences, selection of mutant strains with similar morphology, clustering based on morphology, and study of morphological diversity. In addition to basic research, morphological analyses of yeast cells can also be used in applied research to monitor breeding and fermentation processes and control the fermentation activity of yeast cells.

对出芽酵母进行形态学表型分析有助于极大地阐明基因的功能,加深我们对细胞功能网络的了解。了解细胞形态并进行定量形态分析(QMA)是必要的,但为形态表型分配精确的数值一直是个挑战。为此,我们最近开发了单模态形态数据图像分析管道。如果实验值的分布遵循单模态模式,所有真实值都可以通过应用适当的概率分布从理论上估算出来。这一可靠的管道可进行多种下游分析,包括检测细微的形态差异、选择形态相似的突变株、基于形态的聚类以及形态多样性研究。除基础研究外,酵母细胞形态分析还可用于应用研究,监测育种和发酵过程,控制酵母细胞的发酵活性。
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引用次数: 0
Exploring the antioxidant activity of Fe(III), Mn(III)Mn(II), and Cu(II) compounds in Saccharomyces cerevisiae and Galleria mellonella models of study 探究铁(III)、锰(III)、锰(II)和铜(II)化合物在酿酒酵母和黑僵菌模型中的抗氧化活性
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-12-21 DOI: 10.1093/femsyr/foad052
Larissa M M Mattos, Hyan M Hottum, Daniele C Pires, Bruna B Segat, Adolfo Horn, Christiane Fernandes, Marcos D Pereira
Reactive oxygen species (ROS) are closely related to oxidative stress, aging, and the onset of human diseases. To mitigate ROS-induced damages, extensive research has focused on examining the antioxidative attributes of various synthetic/natural substances. Coordination compounds serving as synthetic antioxidants have emerged as a promising approach to attenuate ROS toxicity. Herein, we investigated the antioxidant potential of a series of Fe(III) (1), Mn(III)Mn(II) (2) and Cu(II) (3) coordination compounds synthesized with the ligand N-(2-hydroxybenzyl)-N-(2-pyridylmethyl)[(3-chloro)(2-hydroxy)]-propylamine in Saccharomyces cerevisiae exposed to oxidative stress. We also assessed the antioxidant potential of these complexes in the alternative model of study, Galleria mellonella. DPPH analysis indicated that these complexes presented moderate antioxidant activity. However, treating Saccharomyces cerevisiae with 1, 2 and 3 increased the tolerance against oxidative stress and extended yeast lifespan. The treatment of yeast cells with these complexes decreased lipid peroxidation and catalase activity in stressed cells, whilst no change in SOD activity was observed. Moreover, these complexes induced the Hsp104 expression. In G. mellonella, complex administration extended larval survival under H2O2 stress and did not affect the insect's life cycle. Our results suggest that the antioxidant potential exhibited by these complexes could be further explored to mitigate various oxidative stress-related disorders.
活性氧(ROS)与氧化应激、衰老和人类疾病的发生密切相关。为了减轻 ROS 引起的损害,大量研究集中于研究各种合成/天然物质的抗氧化特性。作为合成抗氧化剂的配位化合物已成为一种有望减轻 ROS 毒性的方法。在此,我们研究了一系列与配体 N-(2-羟基苄基)-N-(2-吡啶基甲基)[(3-氯)(2-羟基)]-丙胺合成的 Fe(III) (1)、Mn(III)Mn(II) (2) 和 Cu(II) (3) 配位化合物在暴露于氧化应激的酿酒酵母中的抗氧化潜力。我们还评估了这些复合物在另一种研究模型--麦鸡中的抗氧化潜力。DPPH 分析表明,这些复合物具有中等程度的抗氧化活性。不过,用 1、2 和 3 处理酿酒酵母,可提高其对氧化应激的耐受性,并延长酵母的寿命。用这些复合物处理酵母细胞可降低受激细胞的脂质过氧化反应和过氧化氢酶活性,而 SOD 活性则没有变化。此外,这些复合物还能诱导 Hsp104 的表达。在 G. mellonella 中,施用复合物可延长幼虫在 H2O2 胁迫下的存活时间,而且不会影响昆虫的生命周期。我们的研究结果表明,可以进一步开发这些复合物的抗氧化潜力,以缓解各种与氧化应激相关的疾病。
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引用次数: 0
QTL mapping reveals novel genes and mechanisms underlying variations in H2S production during alcoholic fermentation in Saccharomyces cerevisiae QTL 图谱揭示了酿酒酵母在酒精发酵过程中产生 H2S 的新基因和新机制
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-12-16 DOI: 10.1093/femsyr/foad050
Irene De Guidi, Céline Serre, Jessica Noble, Anne Ortiz-Julien, Bruno Blondin, Jean-Luc Legras
Saccharomyces cerevisiae requirement for reduced sulfur to synthesise methionine and cysteine during alcoholic fermentation, is mainly fulfilled through the sulfur assimilation pathway. S. cerevisiae reduces sulfate into sulfur dioxide (SO2) and sulfide (H2S), whose overproduction is a major issue in winemaking, due to its negative impact on wine aroma. The amount of H2S produced is highly strain-specific and also depends on SO2 concentration, often added to grape must. Applying a Bulk Segregant Analysis to a 96 strain-progeny derived from two strains with different abilities to produce H2S, and comparing allelic frequencies along the genome of pools of segregants producing contrasting H2S quantities, we identified two causative regions involved in H2S production in the presence of SO2. A functional genetic analysis allowed the identification of variants in four genes able to impact H2S formation, viz; ZWF1, ZRT2, SNR2 and YLR125W, and involved in functions and pathways not associated with sulfur metabolism until now. These data point out that, in wine fermentation conditions, redox status and zinc homeostasis are linked to H2S formation while providing new insights into the regulation of H2S production, and a new vision of the interplay between the sulfur assimilation pathway and cell metabolism.
酿酒酵母(Saccharomyces cerevisiae)在酒精发酵过程中需要还原硫来合成蛋氨酸和半胱氨酸,这主要是通过硫同化途径来实现的。葡萄酿酒酵母将硫酸盐还原成二氧化硫(SO2)和硫化物(H2S),由于其对葡萄酒香气的负面影响,过量产生硫化物是酿酒过程中的一个主要问题。H2S 的产生量因菌株而异,也取决于二氧化硫的浓度,通常会添加到葡萄汁中。通过对产生 H2S 能力不同的两个菌株所产生的 96 个菌株后代进行大量分离分析,并比较产生 H2S 数量截然不同的分离株基因组的等位基因频率,我们确定了在 SO2 存在的情况下产生 H2S 的两个致病区域。通过功能基因分析,我们发现了四个能够影响 H2S 形成的基因变异,即 ZWF1、ZRT2、SNR2 和 YLR125W,它们参与了迄今为止与硫代谢无关的功能和途径。这些数据表明,在葡萄酒发酵条件下,氧化还原状态和锌平衡与 H2S 的形成有关,同时为 H2S 生成的调控提供了新的见解,并为硫同化途径与细胞代谢之间的相互作用提供了新的视角。
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引用次数: 0
My Journey with Yeast. 我的酵母之旅。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-06-22 DOI: 10.1093/femsyr/foad035
Charles Abbas

Advances in yeast biotechnology rely on the application of knowledge gained using modern biotechnological tools to harness the metabolic repertoire of various yeast genera that have been studied in detail. In my work, I have attempted to combine knowledge gained from academic research with industrial knowhow in practical cost-effective ways to scale up commercial yeast fermentations from one hundred thousand to greater than one million liters. Among the processes I scaled up and/or optimized to production scale include biofuels, chemicals, food and feed additives. During a long industrial career that spanned over three decades, I leveraged what was often very challenging work with many academic, government, and industrial scientists engaging them in collaborative research to ensure a successful outcome. Many of these collaborators responded in kind and are part of this narrative. In many ways, my journey was also theirs. However, I acquired the scientific foundation or starting point much earlier during my undergraduate studies learning from great professors that helped me understand the complexity of science while convincing me of the value of pursuing research as a career.

酵母生物技术的进步依赖于利用现代生物技术工具获得的知识来利用已经详细研究过的各种酵母属的代谢库。在我的工作中,我试图将从学术研究中获得的知识与工业技术结合起来,以实际经济有效的方式将商业酵母发酵从10万升扩大到100万升以上。在我扩大和/或优化生产规模的过程中,包括生物燃料、化学品、食品和饲料添加剂。在长达30多年的工业生涯中,我利用了许多学术、政府和工业科学家参与合作研究的挑战性工作,以确保取得成功。这些合作者中的许多人都以同样的方式回应了我,并成为了这个故事的一部分。在很多方面,我的旅程也是他们的。然而,在我的本科学习中,我更早地获得了科学基础或起点,从伟大的教授那里学习,帮助我理解科学的复杂性,同时让我相信追求研究作为一种职业的价值。
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引用次数: 0
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FEMS yeast research
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