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Metabolic reconstruction of the human pathogen Candida auris: using a cross-species approach for drug target prediction. 人类病原体耳念珠菌的代谢重建:使用跨物种方法预测药物靶点。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad045
Romeu Viana, Tiago Carreiro, Diogo Couceiro, Oscar Dias, Isabel Rocha, Miguel Cacho Teixeira

Candida auris is an emerging human pathogen, associated with antifungal drug resistance and hospital candidiasis outbreaks. In this work, we present iRV973, the first reconstructed Genome-scale metabolic model (GSMM) for C. auris. The model was manually curated and experimentally validated, being able to accurately predict the specific growth rate of C. auris and the utilization of several sole carbon and nitrogen sources. The model was compared to GSMMs available for other pathogenic Candida species and exploited as a platform for cross-species comparison, aiming the analysis of their metabolic features and the identification of potential new antifungal targets common to the most prevalent pathogenic Candida species. From a metabolic point of view, we were able to identify unique enzymes in C. auris in comparison with other Candida species, which may represent unique metabolic features. Additionally, 50 enzymes were identified as potential drug targets, given their essentiality in conditions mimicking human serum, common to all four different Candida models analysed. These enzymes represent interesting drug targets for antifungal therapy, including some known targets of antifungal agents used in clinical practice, but also new potential drug targets without any human homolog or drug association in Candida species.

耳念珠菌是一种新出现的人类病原体,与抗真菌耐药性和医院念珠菌感染暴发有关。在这项工作中,我们提出了iRV973,这是第一个重建的金黄色葡萄球菌基因组级代谢模型(GSMM)。该模型经过人工策划和实验验证,能够准确预测金合欢的比生长速率以及几种唯一碳和氮来源的利用率。将该模型与可用于其他致病性念珠菌物种的GSMM进行比较,并将其作为跨物种比较的平台,旨在分析其代谢特征,并确定最常见的致病性念珠菌常见的潜在新抗真菌靶标。从代谢的角度来看,与其他念珠菌物种相比,我们能够在耳念珠菌中鉴定出独特的酶,这可能代表了独特的代谢特征。此外,考虑到50种酶在模拟人类血清的条件下的重要性,它们被确定为潜在的药物靶点,这是所分析的所有四种不同念珠菌模型所共有的。这些酶代表了抗真菌治疗的有趣药物靶点,其中一些是临床实践中使用的抗真菌药物的已知靶点,但其他新的潜在药物靶点也很突出,在念珠菌中没有任何人类同源物或药物关联。
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引用次数: 0
Correction to: The prevalence of killer yeasts and double-stranded RNAs in the budding yeast Saccharomyces cerevisiae. 更正:芽殖酵母中致命酵母和双链 RNA 的普遍性。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad051
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引用次数: 0
Optimizing the balance between heterologous acetate- and CO2-reduction pathways in anaerobic cultures of Saccharomyces cerevisiae strains engineered for low-glycerol production. 优化用于低甘油生产的酿酒酵母菌株厌氧培养中异源乙酸盐和CO2还原途径之间的平衡。
IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad048
Aafke C A van Aalst, Ellen H Geraats, Mickel L A Jansen, Robert Mans, Jack T Pronk

In anaerobic Saccharomyces cerevisiae cultures, NADH (reduced form of nicotinamide adenine dinucleotide)-cofactor balancing by glycerol formation constrains ethanol yields. Introduction of an acetate-to-ethanol reduction pathway based on heterologous acetylating acetaldehyde dehydrogenase (A-ALD) can replace glycerol formation as 'redox-sink' and improve ethanol yields in acetate-containing media. Acetate concentrations in feedstock for first-generation bioethanol production are, however, insufficient to completely replace glycerol formation. An alternative glycerol-reduction strategy bypasses the oxidative reaction in glycolysis by introducing phosphoribulokinase (PRK) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO). For optimal performance in industrial settings, yeast strains should ideally first fully convert acetate and, subsequently, continue low-glycerol fermentation via the PRK-RuBisCO pathway. However, anaerobic batch cultures of a strain carrying both pathways showed inferior acetate reduction relative to a strain expressing only the A-ALD pathway. Complete A-ALD-mediated acetate reduction by a dual-pathway strain, grown anaerobically on 50 g L-1 glucose and 5 mmol L-1 acetate, was achieved upon reducing PRK abundance by a C-terminal extension of its amino acid sequence. Yields of glycerol and ethanol on glucose were 55% lower and 6% higher, respectively, than those of a nonengineered reference strain. The negative impact of the PRK-RuBisCO pathway on acetate reduction was attributed to sensitivity of the reversible A-ALD reaction to intracellular acetaldehyde concentrations.

在厌氧酿酒酵母培养物中,通过甘油形成的NADH辅因子平衡限制了乙醇产量。基于异源乙酰化乙醛脱氢酶(A-ALD)的乙酸盐-乙醇还原途径的引入可以取代甘油作为“氧化还原库”的形成,并提高含乙酸盐介质中的乙醇产量。然而,第一代生物乙醇生产原料中的乙酸盐浓度不足以完全取代甘油的形成。另一种甘油还原策略通过引入磷酸二激酶(PRK)和核酮糖-1,5-二磷酸羧化酶/加氧酶(RuBisCO)绕过糖酵解中的氧化反应。为了在工业环境中获得最佳性能,理想情况下,酵母菌株应首先完全转化乙酸盐,然后通过PRK-RuBisCO途径继续低甘油发酵。然而,与仅表达a-ALD途径的菌株相比,携带两种途径的菌株的厌氧分批培养显示出较差的乙酸盐还原。在50 g L-1葡萄糖和5 mmol L-1乙酸盐上厌氧生长的双途径菌株通过其氨基酸序列的C末端延伸降低PRK丰度后,实现了A-ALD介导的乙酸盐的完全还原。甘油和乙醇对葡萄糖的产量分别比非工程参考菌株低55%和高6%。PRK-RuBisCO途径对乙酸盐还原的负面影响归因于可逆A-ALD反应对细胞内乙醛浓度的敏感性。
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引用次数: 0
Current trends and future perspective of probiotic yeasts research in Indonesia. 印尼益生菌研究现状及未来展望
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad013
Rika Indri Astuti, Muhammad Eka Prastya, Rahayu Wulan, Khairul Anam, Anja Meryandini

Indonesia is a mega biodiversity country with various local wisdom, including the enormous variety of fermented foods and beverages. Indonesian researchers have conducted an intensive study to understand the diversity of microbes on those fermented products, one of which shows probiotic properties. Compared to that lactic acid bacteria, the study on probiotic yeasts is less explored. Probiotic yeast isolates are commonly isolated from traditional Indonesian fermented products. Saccharomyces, Pichia, and Candida are among Indonesia's most popular genera of probiotic yeasts, primarily applied in poultry and human health. The exploration of functional probiotic characteristics, such as antimicrobial, antifungal, antioxidant, and immunomodulator, has been widely reported from these local probiotic yeast strains. In vivo studies in a model organism such as mice conclude the prospective functional probiotic characteristics of the yeast isolates. Employment of current technology, such as omics, is essential in elucidating those functional properties. Advanced research and development of probiotic yeasts in Indonesia are gaining significant attention currently. For instance, probiotic yeasts-mediated fermentation in the production of kefir and kombucha are among the trend with promising economic value. The future trends of probiotic yeasts research in Indonesia are discussed in this review to give valuable sight into the application of indigenous probiotic yeasts in various fields.

印度尼西亚是一个生物多样性丰富的国家,拥有各种各样的地方智慧,包括种类繁多的发酵食品和饮料。印度尼西亚的研究人员进行了深入的研究,以了解这些发酵产品上微生物的多样性,其中一种产品显示出益生菌的特性。与乳酸菌相比,益生菌酵母的研究较少。分离的益生菌酵母通常是从传统的印度尼西亚发酵产品中分离出来的。酵母菌、毕赤酵母和念珠菌是印度尼西亚最受欢迎的益生菌酵母属,主要用于家禽和人类健康。对益生菌抗菌、抗真菌、抗氧化、免疫调节等功能特性的探索已被广泛报道。在模型生物(如小鼠)的体内研究总结了酵母分离物的潜在功能益生菌特性。使用当前的技术,如组学,对于阐明这些功能特性是必不可少的。目前,印尼对益生菌酵母的先进研究和开发受到了极大的关注。例如,益生菌酵母介导发酵生产克菲尔和康普茶是具有良好经济价值的趋势之一。本文对印尼益生菌研究的未来发展趋势进行了展望,以期对本土益生菌在各个领域的应用提供有益的参考。
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引用次数: 1
PCR-based gene targeting in Hanseniaspora uvarum. 基于pcr的兔痘菌基因靶向研究。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad034
Jennifer Badura, Niël van Wyk, Kerstin Zimmer, Isak S Pretorius, Christian von Wallbrunn, Jürgen Wendland

Lack of gene-function analyses tools limits studying the biology of Hanseniaspora uvarum, one of the most abundant yeasts on grapes and in must. We investigated a rapid PCR-based gene targeting approach for one-step gene replacement in this diploid yeast. To this end, we generated and validated two synthetic antibiotic resistance genes, pFA-hygXL and pFA-clnXL, providing resistance against hygromycin and nourseothricin, respectively, for use with H. uvarum. Addition of short flanking-homology regions of 56-80 bp to these selection markers via PCR was sufficient to promote gene targeting. We report here the deletion of the H. uvarum LEU2 and LYS2 genes with these marker genes via two rounds of consecutive transformations, each resulting in the generation of auxotrophic strains (leu2/leu2; lys2/lys2). The hereby constructed leucine auxotrophic leu2/leu2 strain was subsequently complemented in a targeted manner, thereby further validating this approach. PCR-based gene targeting in H. uvarum was less efficient than in Saccharomyces cerevisiae. However, this approach, combined with the availability of two marker genes, provides essential tools for directed gene manipulations in H. uvarum.

缺乏基因功能分析工具限制了对葡萄和木薯上最丰富的酵母之一——葡萄芽孢酵母的生物学研究。我们研究了一种基于pcr的快速基因靶向方法,用于二倍体酵母的一步基因替换。为此,我们合成并验证了两个合成的抗生素耐药基因pFA-hygXL和pFA-clnXL,分别提供了对潮霉素和诺斯库霉素的耐药,用于uvarum。通过PCR在这些选择标记上添加56 ~ 80bp的短侧同源区域,足以促进基因靶向。我们在这里报道,通过两轮连续转化,uvarum的LEU2和LYS2基因与这些标记基因的缺失,每一轮都导致产生营养不良菌株(LEU2 / LEU2;lys2 / lys2)。由此构建的亮氨酸营养不良leu2/leu2菌株随后被有针对性地补充,从而进一步验证了该方法。在酿酒酵母中,基于pcr的基因靶向效率较低。然而,这种方法与两个标记基因的可用性相结合,为uvarum的定向基因操作提供了必要的工具。
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引用次数: 1
Red yeasts and their carotenogenic enzymes for microbial carotenoid production. 用于微生物类胡萝卜素生产的红酵母及其生胡萝卜素酶。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foac063
Akaraphol Watcharawipas, Weerawat Runguphan

Carotenoids are C40 isoprene-based compounds with significant commercial interests that harbor diverse bioactivities. Prominent examples of carotenoids are beta-carotene, a precursor to vitamin A essential for proper eye health, and lycopene and astaxanthin, powerful antioxidants implicated in preventing cancers and atherosclerosis. Due to their benefits to human health, the market value for carotenoids is rapidly increasing and is projected to reach USD 1.7 billion by 2025. However, their production now relies on chemical synthesis and extraction from plants that pose risks to food management and numerous biological safety issues. Thus, carotenoid production from microbes is considered a promising strategy for achieving a healthy society with more sustainability. Red yeast is a heterogeneous group of basidiomycetous fungi capable of producing carotenoids. It is a critical source of microbial carotenoids from low-cost substrates. Carotenogenic enzymes from red yeasts have also been highly efficient, invaluable biological resources for biotechnological applications. In this minireview, we focus on red yeast as a promising source for microbial carotenoids, strain engineering strategies for improving carotenoid production in red yeasts, and potential applications of carotenogenic enzymes from red yeasts in conventional and nonconventional yeasts.

类胡萝卜素是基于C40异戊二烯的化合物,具有重要的商业价值,具有多种生物活性。类胡萝卜素的典型例子是-胡萝卜素,它是维生素a的前体,对眼睛健康至关重要,还有番茄红素和虾青素,它们是有效的抗氧化剂,可以预防癌症和动脉粥样硬化。由于类胡萝卜素对人类健康有益,其市场价值正在迅速增长,预计到2025年将达到17亿美元。然而,它们的生产现在依赖于化学合成和从植物中提取,这对食品管理和许多生物安全问题构成了风险。因此,从微生物中生产类胡萝卜素被认为是实现更具可持续性的健康社会的一种有希望的策略。红曲是一种能够产生类胡萝卜素的担子菌真菌。它是从低成本基质中提取微生物类胡萝卜素的重要来源。从红酵母中提取的胡萝卜素酶也是生物技术应用的高效、宝贵的生物资源。在这篇综述中,我们重点介绍了红酵母作为微生物类胡萝卜素的有前途的来源,菌株工程策略,以提高红酵母类胡萝卜素的产量,以及红酵母的胡萝卜素酶在常规和非常规酵母中的潜在应用。
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引用次数: 3
Application of the fission yeast Schizosaccharomyces pombe in human nutrition. 裂糖酵母在人体营养中的应用。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foac064
Ee Sin Chen

Fission yeast Schizosaccharomyces pombe (S. pombe) is renowned as a powerful genetic model for deciphering cellular and molecular biological phenomena, including cell division, chromosomal events, stress responses, and human carcinogenesis. Traditionally, Africans use S. pombe to ferment the beer called 'Pombe', which continues to be consumed in many parts of Africa. Although not as widely utilized as the baker's yeast Saccharomyces cerevisiae, S. pombe has secured several niches in the food industry for human nutrition because of its unique metabolism. This review will explore three specific facets of human nutrition where S. pombe has made a significant impact: namely, in wine fermentation, animal husbandry and neutraceutical supplementation coenzyme Q10 production. Discussions focus on the current gaps in these areas, and the potential research advances useful for addressing future challenges. Overall, gaining a better understanding of S. pombe metabolism will strengthen production in these areas and potentially spearhead novel future applications.

分裂酵母pombe Schizosaccharomyces (S. pombe)被认为是一种强大的遗传模型,可以破译细胞和分子生物学现象,包括细胞分裂、染色体事件、应激反应和人类癌变。传统上,非洲人用S. pombe来发酵一种名为“pombe”的啤酒,这种啤酒在非洲的许多地方仍然被消费。虽然不像面包酵母那样广泛使用,但由于其独特的新陈代谢,s.p ombe在食品工业中为人类营养提供了几个利基。本文将探讨pombe在葡萄酒发酵、畜牧业和中性保健品辅酶Q10生产等三个方面对人类营养的影响。讨论的重点是这些领域目前的差距,以及对解决未来挑战有用的潜在研究进展。总的来说,更好地了解S. pombe代谢将加强这些领域的生产,并有可能引领新的未来应用。
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引用次数: 1
Identification of European isolates of the lager yeast parent Saccharomyces eubayanus. 欧洲大酵母亲本真bayanus Saccharomyces鉴定。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2022-12-07 DOI: 10.1093/femsyr/foac053
Sean A Bergin, Stephen Allen, Conor Hession, Eoin Ó Cinnéide, Adam Ryan, Kevin P Byrne, Tadhg Ó Cróinín, Kenneth H Wolfe, Geraldine Butler

Lager brewing first occurred in Bavaria in the 15th century, associated with restrictions of brewing to colder months. The lager yeast, Saccharomyces pastorianus, is cold tolerant. It is a hybrid between Saccharomyces cerevisiae and Saccharomyces eubayanus, and has been found only in industrial settings. Natural isolates of S. eubayanus were first discovered in Patagonia 11 years ago. They have since been isolated from China, Tibet, New Zealand, and North America, but not from Europe. Here, we describe the first European strains UCD646 and UCD650, isolated from a wooded area on a university campus in Dublin, Ireland. We generated complete chromosome level assemblies of both genomes using long- and short-read sequencing. The UCD isolates belong to the Holarctic clade. Genome analysis shows that isolates similar to the Irish strains contributed to the S. eubayanus component of S. pastorianus, but isolates from Tibet made a larger contribution.

啤酒酿造最早出现在15世纪的巴伐利亚,当时酿造的时间限制在较冷的月份。窖藏酵母,酵母,是耐寒的。它是酿酒酵母菌和真bayanus酵母菌的杂交品种,只在工业环境中被发现。真巴bayanus的天然分离株是11年前在巴塔哥尼亚首次发现的。它们已经从中国、西藏、新西兰和北美分离出来,但没有从欧洲分离出来。在这里,我们描述了第一个欧洲菌株UCD646和UCD650,从爱尔兰都柏林一所大学校园的树林中分离出来。我们使用长读和短读测序产生了两个基因组的完整染色体水平组装。UCD分离株属于全北极分支。基因组分析表明,与爱尔兰菌株相似的分离株贡献了巴斯德多利葡萄球菌的真bayanus成分,但来自西藏的分离株贡献更大。
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引用次数: 7
Saccharomyces eubayanus-a tale of endless mysteries. 真酵母——一个充满无尽谜团的故事。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2022-12-07 DOI: 10.1093/femsyr/foac058
José Paulo Sampaio
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引用次数: 1
Functional diversity and plasticity in the sugar preferences of Saccharomyces MALT transporters in domesticated yeasts. 驯化酵母中酵母菌MALT转运体对糖偏好的功能多样性和可塑性。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2022-12-07 DOI: 10.1093/femsyr/foac055
Haruyo Hatanaka, Hiromi Toyonaga, Yukiko Ishida, Eiichi Mizohata, Eiichiro Ono

Maltose and maltotriose, together with glucose, are the major carbohydrates found in malts. Thus, brewing yeasts grown in malt-based brewing processes with serial re-pitching have likely increased their ability to uptake these sugars during domestication by modulating the expression and copy number of maltose transporter genes (MALT, also known as Malx1). However, the molecular basis for and structural insights into the sugar preferences of MALT proteins remain to be elucidated. Here we report the functional evaluation of two novel Saccharomyces cerevisiae MALT proteins, ScMalt#2p and ScMalt#5p, from industrial brewing yeasts, focusing on their maltose and maltotriose preferences. Structural models of the MALT proteins generated by AlphaFold2 and functional analyses of substitution mutants revealed that a very small number of amino acid residues in two spatially adjacent transmembrane helixes, TMH7 and TMH11, appear to be crucial for sugar preference. Thus, subtle conformational alterations conferred by a small number of amino acid polymorphisms within MALTs would contribute to the adaptation of domesticated brewing yeasts to the constrained carbohydrate environment of industrial wort during beer brewing.

麦芽糖、麦芽糖和葡萄糖是麦芽中主要的碳水化合物。因此,在以麦芽为基础的酿造过程中生长的酿造酵母在驯化过程中通过调节麦芽糖转运基因(MALT,也称为Malx1)的表达和拷贝数,可能增加了它们摄取这些糖的能力。然而,MALT蛋白的糖偏好的分子基础和结构见解仍有待阐明。本文报道了从工业酿造酵母中提取的两种新型酿酒酵母MALT蛋白ScMalt#2p和ScMalt#5p的功能评价,重点研究了它们对麦芽糖和麦芽糖的偏好。由AlphaFold2生成的MALT蛋白的结构模型和对替代突变体的功能分析表明,在两个空间相邻的跨膜螺旋TMH7和TMH11中,极少量的氨基酸残基似乎对糖偏好至关重要。因此,在啤酒酿造过程中,由少量氨基酸多态性引起的细微构象改变将有助于驯化酿酒酵母适应工业麦芽汁的有限碳水化合物环境。
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引用次数: 0
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FEMS yeast research
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