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Metabolic engineering of Mucor circinelloides to improve astaxanthin production. 通过对环褐藻(Mucor circinelloides)进行代谢工程改造,提高虾青素的产量。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-02 DOI: 10.1007/s11274-024-04181-x
Tahira Naz, Tariq Saeed, Samee Ullah, Yusuf Nazir, Molalign Assefa, Qing Liu, Zhaosen Fan, Hassan Mohamed, Yuanda Song

Astaxanthin is a bioactive natural pigment with antioxidant properties. It has extensive applications within the industrial sector as well as in human and animal health. Mucor circinelloides is a zygomycete fungus that accumulates β-carotene as the main carotenoid compound. M. circinelloides is a well-known model organism among Mucorales for studying carotenogenesis in fungi, which makes it a promising candidate for the biotechnological production of carotenoids. In this study, β-carotene hydroxylase (crtR-B) and ketolase (bkt) genes (codon-optimized) were coexpressed from Haematococcus pluvialis in M. circinelloides using two potent promoters gpd1 and zrt1 respectively to generate an astaxanthin-producing biofactory. Following 72 h of cultivation, the recombinant M. circinelloides Mc-57 obtained in this study produced 135 ± 8 µg/g of astaxanthin. This is the highest reported amount in M. circinelloides to date. The mRNA levels of crtR-B and bkt in Mc-57 were assayed using RT-qPCR. These levels showed a 5.7-fold increase at 72 h and a 5.5-fold increase at 24 h, respectively, compared to the control strain. This demonstrated the successful overexpression of both genes, which correlated with the production of astaxanthin in the Mc-57. Moreover, the addition of glutamate (2 g/L) and mevalonate (15 mM) resulted in an increase in astaxanthin production in the recombinant strain. The results showed that the combined addition of these metabolic precursors resulted in 281 ± 20 µg/g of astaxanthin, which is 2.08-fold higher than the control medium (135 ± 8 µg/g). The addition of metabolic precursors also positively impacted the biomass growth of Mc-57, reaching 11.2 ± 0.57 g/L compared to 9.1 ± 0.23 g/L (control medium). The study successfully addressed the challenge of balancing the accumulation of astaxanthin with biomass growth, which has been regarded as common bottleneck in the metabolic engineering of microbial cells. The development of a recombinant fungal strain of M. circinelloides not only increased astaxanthin content. Additionally, it provided a foundation for further improvement of the biotechnological production of astaxanthin in M. circinelloides.

虾青素是一种具有生物活性的天然色素,具有抗氧化特性。它在工业领域以及人类和动物健康方面有着广泛的应用。Mucor circinelloides 是一种子囊真菌,其主要类胡萝卜素化合物是β-胡萝卜素。环翅袋霉是真菌中研究类胡萝卜素生成的著名模式生物,这使其成为生物技术生产类胡萝卜素的理想候选者。在这项研究中,利用两个强效启动子 gpd1 和 zrt1,分别将血球菌的β-胡萝卜素羟化酶(crtR-B)和酮酶(bktolase)基因(经过密码子优化)在环丝菌中共表达,生成了一个生产虾青素的生物工厂。经过 72 小时的培养,本研究中获得的重组环带藻 M. Mc-57 产生了 135 ± 8 µg/g 的虾青素。这是迄今为止报道的最高数量。使用 RT-qPCR 对 Mc-57 中 crtR-B 和 bkt 的 mRNA 水平进行了检测。与对照菌株相比,72 小时和 24 小时的 mRNA 水平分别增加了 5.7 倍和 5.5 倍。这表明这两个基因的成功过表达与 Mc-57 产生虾青素有关。此外,添加谷氨酸(2 g/L)和甲羟戊酸(15 mM)可提高重组菌株的虾青素产量。结果显示,联合添加这些代谢前体后,虾青素的产量为 281 ± 20 µg/g,比对照培养基(135 ± 8 µg/g)高出 2.08 倍。添加代谢前体还对 Mc-57 的生物量增长产生了积极影响,使其达到 11.2 ± 0.57 克/升,而对照培养基为 9.1 ± 0.23 克/升。这项研究成功地解决了虾青素积累与生物量增长之间的平衡难题,而这一直被认为是微生物细胞代谢工程中的常见瓶颈。环带藻真菌重组菌株的开发不仅提高了虾青素的含量,还为其在生物质中的积累奠定了基础。此外,它还为进一步改进环带褐藻虾青素的生物技术生产奠定了基础。
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引用次数: 0
Exploration of the advantages of targeted isolation of deep-sea microorganisms and genetically engineered strains. 探索定向分离深海微生物和基因工程菌株的优势。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-02 DOI: 10.1007/s11274-024-04177-7
MengYao Liu, Yali Feng, Haoran Li, Yisong Yao, Yufeng Cui, Jianwei Wang

Oil, mineral processing and environmental restoration can be dangerous processes. Attempts are often made to apply microorganisms to reduce the risks, but the adaptability of terrestrial organisms is often weak. Although genetically engineered strains can improve their environmental adaptability through targeted modification, there are problems such as metabolite accumulation, poor plasmid stability and potential pathogenicity. Screening of extremophiles from the natural environment has become an inevitable choice. The special environment in the deep sea (high pressure, low temperature, low nutrition, high salinity) is a natural place for extremophiles to grow and survive, thus screening of extremophiles from the deep sea is conducive to the green and sustainable development of industry. In this paper, the application status and problems of genetically engineered strains are reviewed based on the microorganisms needed for extreme industry. This paper focuses on the application status and advantages of deep-sea microorganisms. It is found that their advantages are strong adaptability, stable gene, friendly environment, simple and convenient technology (compared with genetic engineering), which has a broad industry processes application prospect. This review broadens the scope of microbial applications.

石油、矿物加工和环境恢复可能是危险的过程。人们常常试图利用微生物来降低风险,但陆生生物的适应性往往很弱。虽然基因工程菌株可以通过定向改造提高环境适应能力,但也存在代谢物积累、质粒稳定性差和潜在致病性等问题。从自然环境中筛选嗜极生物已成为必然选择。深海的特殊环境(高压、低温、低营养、高盐度)是嗜极端生物生长和生存的天然场所,因此从深海筛选嗜极端生物有利于产业的绿色可持续发展。本文根据极端工业所需的微生物,综述了基因工程菌株的应用现状和存在的问题。本文重点探讨了深海微生物的应用现状和优势。研究发现,其优势在于适应性强、基因稳定、环境友好、技术简单方便(与基因工程相比),具有广阔的工业过程应用前景。这篇综述拓宽了微生物的应用范围。
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引用次数: 0
Deletion of both anaerobic regulator genes fnr and narL compromises the colonization of Salmonella Typhimurium in mice model. 缺失厌氧调节基因 fnr 和 narL 会影响鼠伤寒沙门氏菌的定植。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-02 DOI: 10.1007/s11274-024-04179-5
Swagatika Priyadarsini, Pashupathi Mani, Rohit Singh, K C Nikhil, Pravas Ranjan Sahoo, M Kesavan, Meeta Saxena, Monalisa Sahoo, Mohini Saini, Ajay Kumar

Salmonella Typhimurium (STM), a zoonotic pathogen, can adjust its metabolic pathway according to the variations in the partial pressure of atmospheric oxygen and nitrate via fumarate nitrate reductase regulator (Fnr) and NarL, the response regulator for nitrate reductase. Both Fnr and NarL have been individually reported to be the contributors of virulent phenotypes of STM. Hypoxia along with nitrate-rich environment are prevalent in macrophages and the Salmonella-induced inflammatory lumen of the host's large intestine activates both fnr and narL genes. In this study, the double (fnr and narL) knockout STM showed a synergistic reduction in the swimming (62%), swarming (84%) and biofilm density (86%) phenotypes anaerobically in association with its significant aerobic attenuation. The intracellular replication of the double mutant was reduced by 2.3 logs in chicken monocyte-derived macrophages. Furthermore, the competitive index of the double mutant in liver and spleen was found to be 0.3 and 0.44 respectively at 120 h post-infection (PI) in mice. Surprisingly, no double mutant could be recovered from the infected mouse liver 3 days PI. Histopathological findings showed moderate infiltration of mononuclear cells in the large intestine of mice infected with double mutant, but severe infiltration was seen with the wild-type strain.

鼠伤寒沙门氏菌(STM)是一种人畜共患的病原体,它可以通过富马酸硝酸还原酶调节器(Fnr)和硝酸还原酶的反应调节器 NarL,根据大气氧分压和硝酸分压的变化调整其代谢途径。据报道,Fnr 和 NarL 都是 STM 毒性表型的促成因素。巨噬细胞中普遍存在缺氧和富含硝酸盐的环境,而沙门氏菌诱发的宿主大肠炎症腔能同时激活 fnr 和 NarL 基因。在这项研究中,双(fnr 和 narL)基因敲除的 STM 在厌氧条件下表现出协同减少游动(62%)、成群(84%)和生物膜密度(86%)的表型,同时显著减少需氧条件下的表型。双突变体在鸡单核巨噬细胞中的细胞内复制减少了 2.3 logs。此外,在小鼠感染后 120 小时(PI),双突变体在肝脏和脾脏中的竞争指数分别为 0.3 和 0.44。令人惊讶的是,在感染后 3 天,小鼠肝脏中没有发现双突变体。组织病理学结果显示,感染了双突变株的小鼠大肠中出现了中度的单核细胞浸润,而野生型菌株则出现了严重的单核细胞浸润。
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引用次数: 0
Comparative genomic analysis and characterization of novel high-quality draft genomes from the coal metagenome. 来自煤炭元基因组的新型高质量基因组草案的比较基因组分析和特征描述。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-01 DOI: 10.1007/s11274-024-04174-w
Arunmozhi Bharathi Achudhan, Lilly M Saleena

Coal, a sedimentary rock harbours a complex microbial community that plays a significant role in its formation and characteristics. However, coal metagenome sequencing and studies were less, limiting our understanding of this complex ecosystem. This study aimed to reconstruct high-quality metagenome-assembled genomes (MAGs) from the coal sample collected in the Neyveli mine to explore the unrevealed diversity of the coal microbiome. Using Illumina sequencing, we obtained high-quality raw reads in FASTQ format. Subsequently, de novo assembly and binning with metaWRAP software facilitated the reconstruction of coal MAGs. Quality assessment using CheckM identified 10 High-Quality MAGs (HQ MAGs), 7 medium-quality MAGs (MQ MAGs), and 6 low-quality MAGs (LQ MAGs). Further analysis using GTDB-Tk revealed four HQ MAGs as known species like Dermacoccus abyssi, Sphingomonas aquatilis, Acinetobacter baumannii, and Burkholderia cenocepacia. The remaining six HQ MAGs were classified as Comamonas, Arthrobacter, Noviherbaspirillum, Acidovorax, Oxalicibacterium, and Bordetella and designated as novel genomes by the validation of digital DNA-DNA hybridization (dDDH). Phylogenetic analysis and further pangenome analysis across the phylogenetic groups revealed a similar pattern with a high proportion of cloud genes. We further analysed the functional potential of these MAGs and closely related genomes using COG. The comparative functional genomics revealed that novel genomes are highly versatile, potentially reflecting adaptations to the coal environment. BlastKOALA was used to conduct a detailed analysis of the metabolic pathways associated with the MAGs. This study highlights the comparative genomic analysis of novel coal genomes with their closely related genomes to understand the evolutionary relationships and functional properties.

煤炭是一种沉积岩,其中蕴藏着复杂的微生物群落,对煤炭的形成和特性起着重要作用。然而,煤的元基因组测序和研究较少,限制了我们对这一复杂生态系统的了解。本研究旨在从 Neyveli 煤矿采集的煤炭样本中重建高质量的元基因组组装基因组(MAGs),以探索煤炭微生物组中尚未揭示的多样性。我们利用 Illumina 测序技术获得了 FASTQ 格式的高质量原始读数。随后,使用 metaWRAP 软件进行从头组装和分选,促进了煤微生物群的重建。使用 CheckM 进行的质量评估确定了 10 个高质量 MAGs(HQ MAGs)、7 个中等质量 MAGs(MQ MAGs)和 6 个低质量 MAGs(LQ MAGs)。使用 GTDB-Tk 进行的进一步分析显示,4 个 HQ MAG 为已知物种,如 Dermacoccus abyssi、Sphingomonas aquatilis、Acinetobacter baumannii 和 Burkholderia cenocepacia。其余 6 个 HQ MAG 被归类为 Comamonas、Arthrobacter、Noviherbaspirillum、Acidovorax、Oxalicibacterium 和 Bordetella,并通过数字 DNA-DNA 杂交(dDDH)验证被指定为新基因组。系统发育分析和对各系统发育群的进一步庞基因组分析显示了类似的模式,云基因的比例很高。我们利用 COG 进一步分析了这些 MAG 和密切相关基因组的功能潜力。比较功能基因组学发现,新基因组具有高度多功能性,可能反映了对煤环境的适应性。利用 BlastKOALA 对与 MAGs 相关的代谢途径进行了详细分析。这项研究强调了对新型煤基因组及其近缘基因组进行比较基因组分析,以了解其进化关系和功能特性。
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引用次数: 0
Exploring Ganoderma lucidum: morphology, cultivation and market potential. 灵芝探索:形态、栽培和市场潜力。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-29 DOI: 10.1007/s11274-024-04180-y
Pratibha Thakur, Sonali Khanal, Ashwani Tapwal, Dinesh Kumar, Rachna Verma, Poonam Chauhan, Neha Sharma

Ganoderma lucidum, known as the "mushroom of immortality," is a white rot fungus renowned for its medicinal properties, attributed to its bioactive compounds. Although species with similar morphological traits to G. lucidum are found across the globe, precise identification is made possible through DNA barcoding and molecular phylogenetic analysis. Global cultivation and wild harvesting of G. lucidum are both done in response to the growing market needs. Artificial cultivation is typically performed on sawdust, but other woody substrates and the wood log method are also employed. This cultivation leverages the fungus's ecological role in converting industrial and agricultural solid wastes into biomass, thereby producing functional food and potential pharmaceutical sources. The review consolidates research on various aspects of, including cultivation methods (sawdust, agricultural waste, wood logs, and submerged fermentation), and the current global market conditions.

灵芝被称为 "长生不老的蘑菇",是一种白腐菌,因其生物活性化合物而具有药用价值。虽然全球各地都有与灵芝形态特征相似的物种,但通过 DNA 条形码和分子系统发育分析,可以对其进行精确鉴定。全球栽培和野生采摘绿巨人都是为了满足不断增长的市场需求。人工栽培通常在锯屑上进行,但也采用其他木质基质和原木方法。这种栽培充分利用了真菌将工业和农业固体废弃物转化为生物质的生态作用,从而生产出功能性食品和潜在的药物来源。本综述整合了各方面的研究,包括栽培方法(锯末、农业废料、原木和浸没发酵)以及当前的全球市场状况。
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引用次数: 0
Co-culture systems of microalgae and heterotrophic microorganisms: applications in bioproduction and wastewater treatment and elucidation of mutualistic interactions. 微藻和异养微生物共培养系统:在生物生产和废水处理中的应用,以及相互影响的阐释。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-26 DOI: 10.1007/s11274-024-04173-x
Miiku Takahashi, Ryosuke Yamada, Takuya Matsumoto, Hiroyasu Ogino

In recent years, reducing the concentration of carbon dioxide in the atmosphere has become an important issue. Microalgae have a higher photosynthetic efficiency and growth rate than higher plants; thus, biological carbon dioxide fixation using microalgae is attracting particular attention as an efficient carbon dioxide fixation method. However, under dilute atmospheric conditions, microalgae exhibit lower growth rates and reduced carbon dioxide fixation efficiency. In recent years, technology that can efficiently fix carbon dioxide, even in the atmosphere, using a microalgae co-culture system that co-cultivates microalgae and heterotrophic microorganisms has attracted attention. In such a co-culture system, it is believed that a mutualistic relationship is established between microorganisms through the exchange of various compounds. This review focuses on the application of a co-culture system of microalgae and heterotrophic microorganisms for bioproduction and wastewater treatment. In addition, research to elucidate the mutualistic relationships in microalgal co-culture systems using analytical methods that have been widely used in recent years, such as next-generation sequencing technology, is also discussed. In the future, it is expected that the use of microalgae co-culture systems will expand on an industrial scale through the development of key technologies, such as efficient genetic modification techniques for microalgae and their heterotrophic microorganism partners, large-scale cultivation facilities that can efficiently cultivate microalgae, and stable control techniques for co-culture systems using advanced technology.

近年来,降低大气中的二氧化碳浓度已成为一个重要问题。与高等植物相比,微藻具有更高的光合效率和生长速度,因此,作为一种高效的二氧化碳固定方法,利用微藻进行生物二氧化碳固定尤其受到关注。然而,在稀薄的大气条件下,微藻的生长速度较低,二氧化碳的固定效率也会降低。近年来,利用微藻与异养微生物共同培养的微藻共培养系统,即使在大气中也能高效固定二氧化碳的技术备受关注。在这种共培养系统中,微生物之间通过交换各种化合物建立了互利关系。本综述重点介绍微藻与异养微生物共培养系统在生物生产和废水处理中的应用。此外,还讨论了利用近年来广泛应用的分析方法(如新一代测序技术)阐明微藻共培养系统中的互作关系的研究。今后,通过开发微藻及其异养微生物伙伴的高效基因修饰技术、可高效培养微藻的大规模培养设施以及利用先进技术的共培养系统稳定控制技术等关键技术,预计微藻共培养系统的应用将在工业规模上得到扩展。
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引用次数: 0
Research hotspots and emerging trends in growth and development of macrofungi: a bibliometric review based on CiteSpace analysis. 大型真菌生长发育的研究热点和新趋势:基于 CiteSpace 分析的文献计量学综述。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-26 DOI: 10.1007/s11274-024-04168-8
Xueyan Sun, Dongmei Liu, Xihong Zhao

Macrofungi (or mushrooms) are essential for agriculture, food, and ecology. Although research on the growth and development of macrofungi (GDM) can provide insights into their biological characteristics and metabolite synthesis mechanisms, further exploration is needed for a systematic and visual analysis of the current research progress on GDM. To comprehensively understand the research status and development trend of GDM, a total of 545 scientific literature related to GDM in the Web of Science Core Collection database from 2008 to 2024 were searched and selected as research objects. The general information (publication year, country, institution, and cited journal) and the specific information (co-authorship, keyword co-occurrence, and references with strong citation bursts) were mined and visualized in detail based on CiteSpace software. These analyses demonstrated that related research is still fashionable, and China is dominant and influential in this field. More frequent and in-depth cooperation among authors, institutions and regions is conducive to promoting the research on GDM. Additionally, the quantity and cluster analyses of keywords and references demonstrated that medicinal/edible macrofungi and sustainable development (e.g. mushroom substrate reuse) may be research hotspots and will remain popular in the coming years. This research aims to supply researchers with up-to-date knowledge and cutting-edge issues related to GDM by providing a visually appealing representations on quantitative GDM studies.

大型真菌(或蘑菇)是农业、食品和生态学的重要组成部分。虽然对大型真菌(GDM)生长发育的研究可以深入了解其生物学特性和代谢物合成机制,但要系统、直观地分析目前GDM的研究进展,还需要进一步的探索。为全面了解 GDM 的研究现状和发展趋势,本研究在 Web of Science 核心文库中检索并选取了 2008 年至 2024 年与 GDM 相关的科研文献共 545 篇作为研究对象。基于CiteSpace软件,对一般信息(发表年份、国家、机构和被引期刊)和特殊信息(合著者、关键词共现、强引突发参考文献)进行了详细挖掘和可视化分析。这些分析表明,相关研究仍然是一种时尚,而中国在这一领域具有主导地位和影响力。作者、机构和地区之间更频繁、更深入的合作有利于促进 GDM 的研究。此外,对关键词和参考文献的数量和聚类分析表明,药用/食用大型真菌和可持续发展(如蘑菇基质再利用)可能是研究热点,并将在未来几年持续热门。本研究旨在通过提供具有视觉吸引力的 GDM 定量研究表述,为研究人员提供与 GDM 相关的最新知识和前沿问题。
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引用次数: 0
Exploring flagellar contributions to motility and virulence in Arcobacter butzleri. 探索鞭毛对 Arcobacter butzleri 运动和毒性的贡献。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-26 DOI: 10.1007/s11274-024-04175-9
Raquel Santos, Cristiana Mateus, Mónica Oleastro, Susana Ferreira

Flagella is a well-known bacterial structure crucial for motility, which also plays pivotal roles in pathogenesis. Arcobacter butzleri, an enteropathogen, possesses a distinctive polar flagellum whose functional aspects remain largely unexplored. Upon investigating the factors influencing A. butzleri motility, we uncovered that environmental conditions like temperature, oxygen levels, and nutrient availability play a significant role. Furthermore, compounds that are found in human gut, such as short-chain fatty acids, mucins and bile salts, have a role in modulating the motility, and in turn, the pathogenicity of A. butzleri. Further investigation demonstrated that A. butzleri ΔflaA mutant showed a reduction in motility with a close to null average velocity, as well as a reduction on biofilm formation. In addition, compared with the wild-type, the ΔflaA mutant showed a decreased ability to invade Caco-2 cells and to adhere to mucins. Taken together, our findings support the role of environmental conditions and gut host associated compounds influencing key physiological aspects of the gastrointestinal pathogen A. butzleri, such as motility, and support the role of the flagellum on bacterial virulence.

鞭毛是一种众所周知的细菌结构,对运动至关重要,在致病过程中也发挥着关键作用。肠道致病菌丁兹勒弧杆菌(Arcobacter butzleri)具有独特的极鞭毛,其功能方面的研究在很大程度上仍处于空白。在研究影响丁兹勒弧杆菌运动能力的因素时,我们发现温度、氧气水平和营养供应等环境条件起着重要作用。此外,人类肠道中的化合物,如短链脂肪酸、粘蛋白和胆汁盐,也对丁兹勒氏菌的运动能力有调节作用,进而影响其致病性。进一步的研究表明,丁茨勒氏菌 ΔflaA 突变体的运动能力下降,平均速度接近为零,生物膜的形成也有所减少。此外,与野生型相比,ΔflaA 突变体侵入 Caco-2 细胞和粘附粘蛋白的能力也有所下降。总之,我们的研究结果支持环境条件和肠道宿主相关化合物对胃肠道病原体丁茨勒氏菌关键生理方面(如运动)的影响作用,并支持鞭毛对细菌毒力的作用。
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引用次数: 0
Multi-omics profiling reveals the molecular mechanism of Bifidobacterium animalis BB04 in co-culture with Wickerhamomyces anomalus Y-5 to induce bifidocin A synthesis. 多组学分析揭示了动物双歧杆菌 BB04 与 Wickerhamomyces anomalus Y-5 共培养诱导双歧杆菌素 A 合成的分子机制。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-26 DOI: 10.1007/s11274-024-04172-y
Yangshuo Liu, Rong Nie, Kaisheng Shen, Xinjie Diao, Guorong Liu

Bacteriocin is a kind of natural substance that can effectively inhibit bacteria, but its production usually limited by environment. Co-culture is a strategy to stimulate bacteriocin production. Bifidocin A produced by Bifidobacterium animalis BB04, is a novel bacteriocin with a broad-spectrum antimicrobial active of foodborne bacteria. In order to enhance bifidocin A production, bacteriocin-inducing strains were screened firstly in co-cultivation. Then, the molecular mechanism of co-cultural induction was investigated by transcriptomic and proteomic analysis. Finally, the key inducing metabolites were identified by using targeted metabolomic technology. The results showed that Wickerhamomyces anomalus Y-5 in co-cultivation could significantly enhance bifidocin A production, with a 3.00-fold increase compared to mono-culture. The induction may not depend on direct contact with cells and may instead be attributed to be continuous exposure to inducing substances at specific concentration. In co-cultivation, W. anomalus Y-5 up-regulated Hxk2 and Tap42 to activate Glucose-cAMP and Tor and HOG-MAPK pathway, stimulated the expression of the retrograde gene, produced glutamine and glycerol to maintain activity. During this process, glutamine, inosine, guanosine, adenine, uracil, fumaric acid and pyruvic acid produced by W. anomalus Y-5 could induce the synthesis of bifidocin A. In conclusion, W. anomalus Y-5 in co-cultivation induced the synthesis of bifidocin A by regulating various signaling pathways to produce inducing substances. These findings establish a foundation for high-efficient synthesis of bifidocin A and provide a new perspective into the industrial production of bacteriocin.

细菌素是一种能有效抑制细菌的天然物质,但其产生通常受到环境的限制。共培养是刺激细菌素产生的一种策略。由动物双歧杆菌 BB04 产生的双歧杆菌素 A 是一种新型细菌素,对食源性细菌具有广谱抗菌活性。为了提高双歧杆菌素 A 的产量,首先在联合培养中筛选了细菌素诱导菌株。然后,通过转录组和蛋白质组分析研究了共培养诱导的分子机制。最后,利用靶向代谢组学技术确定了关键诱导代谢物。结果表明,共培养的无柄威克汉姆霉菌 Y-5 能显著提高双歧杆菌素 A 的产量,与单培养相比,产量提高了 3.00 倍。这种诱导可能并不依赖于与细胞的直接接触,而是由于持续暴露于特定浓度的诱导物质。在共培养过程中,W. anomalus Y-5 上调 Hxk2 和 Tap42 以激活 Glucose-cAMP 和 Tor 以及 HOG-MAPK 通路,刺激逆行基因的表达,产生谷氨酰胺和甘油以维持活性。在此过程中,W. anomalus Y-5 产生的谷氨酰胺、肌苷、鸟苷、腺嘌呤、尿嘧啶、富马酸和丙酮酸可诱导双歧杆菌素 A 的合成。这些发现为高效合成双歧杆菌素 A 奠定了基础,并为工业化生产细菌素提供了新的视角。
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引用次数: 0
Integrated proteome and pangenome analysis revealed the variation of microalga Isochrysis galbana and associated bacterial community to 2,6-Di-tert-butyl-p-cresol (BHT) stress. 蛋白质组和泛基因组综合分析揭示了微藻Isochrysis galbana和相关细菌群落对2,6-二叔丁基对甲酚(BHT)胁迫的变化。
IF 4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-24 DOI: 10.1007/s11274-024-04171-z
Linke Guo, Shuangwei Li, Dongle Cheng, Xiao Lu, Xinying Gao, Linlin Zhang, Jianjiang Lu

The phenolic antioxidant 2,6-Di-tert-butyl-p-cresol (BHT) has been detected in various environments and is considered a potential threat to aquatic organisms. Algal-bacterial interactions are crucial for maintaining ecosystem balance and elemental cycling, but their response to BHT remains to be investigated. This study analyzed the physiological and biochemical responses of the microalga Isochrysis galbana and the changes of associated bacterial communities under different concentrations of BHT stress. Results showed that the biomass of I. galbana exhibited a decreasing trend with increasing BHT concentrations up to 40 mg/L. The reduction in chlorophyll, carotenoid, and soluble protein content of microalgal cells was also observed under BHT stress. The production of malondialdehyde and the activities of superoxide dismutase, peroxidase, and catalase were further determined. Scanning electron microscopy analysis revealed that BHT caused surface rupture of the algal cells and loss of intracellular nutrients. Proteomic analysis demonstrated the upregulation of photosynthesis and citric acid cycle pathways as a response to BHT stress. Additionally, BHT significantly increased the relative abundance of specific bacteria in the phycosphere, including Marivita, Halomonas, Marinobacter, and Alteromonas. Further experiments confirmed that these bacteria had the ability to utilize BHT as the sole carbon resource for growth, and genes related to the degradation of phenolic compounds were detected through pangenome analysis.

在各种环境中都发现了酚类抗氧化剂 2,6-二叔丁基对甲酚(BHT),它被认为是对水生生物的一种潜在威胁。藻类与细菌之间的相互作用对维持生态系统平衡和元素循环至关重要,但它们对 BHT 的反应仍有待研究。本研究分析了在不同浓度的 BHT 胁迫下,微藻 Isochrysis galbana 的生理生化反应以及相关细菌群落的变化。结果表明,随着 BHT 浓度的增加,I. galbana 的生物量呈下降趋势,最高达 40 毫克/升。在 BHT 胁迫下,还观察到微藻细胞中叶绿素、类胡萝卜素和可溶性蛋白质含量的减少。还进一步测定了丙二醛的产生以及超氧化物歧化酶、过氧化物酶和过氧化氢酶的活性。扫描电子显微镜分析表明,BHT 导致藻细胞表面破裂,细胞内营养物质流失。蛋白质组分析表明,光合作用和柠檬酸循环途径的上调是对 BHT 胁迫的反应。此外,BHT 还显著增加了植物圈中特定细菌的相对丰度,包括 Marivita、Halomonas、Marinobacter 和 Alteromonas。进一步的实验证实,这些细菌有能力利用 BHT 作为生长的唯一碳资源,并通过泛基因组分析检测到与酚类化合物降解有关的基因。
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World journal of microbiology & biotechnology
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