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Applications of bacteriophages in precision engineering of the human gut microbiome
Pub Date : 2025-01-06 DOI: 10.1016/j.engmic.2025.100189
Xiaoxian Kuang , Juntao Shen , Linggang Zheng , Yi Duan , Yingfei Ma , Elaine Lai-Han Leung , Lei Dai
As our understanding of the role of the gut microbiome in human diseases deepens, precision engineering of the gut microbiome using bacteriophages has gained significant attention. Herein, we review the recent advances in bacteriophage-mediated modulation of the gut microbiome, discuss approaches at the ecological and genetic levels, and summarize the challenges and strategies pertinent to each level of intervention. Drawing on the structural attributes of bacteriophages in the context of precision engineering, we examined the latest developments in the field of phage administration. Gaining a nuanced understanding of microbiome manipulation will yield tailored strategies and technologies. This could revolutionize the prevention and treatment of diseases linked to gut pathogens and offer new avenues for the therapeutic use of bacteriophages.1
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引用次数: 0
BTG13-related metalloenzymes: Atypical non-heme iron-dependent dioxygenases with unusual coordination patterns and catalytic mechanisms
Pub Date : 2025-01-01 DOI: 10.1016/j.engmic.2024.100188
Zhiwei Deng, Zhenbo Yuan, Zhengshan Luo, Yijian Rao
Owing to their diverse coordination patterns and catalytic mechanisms, non-heme iron-dependent dioxygenases catalyze a variety of biochemical reactions involved in the synthesis of numerous natural products and valuable compounds. Recently, we discovered a novel and atypical non-heme iron-dependent dioxygenase, BTG13, that features a unique coordination center consisting of four histidines and a carboxylated lysine (Kcx). This enzyme catalyzes the C–C bond cleavage of anthraquinone through two unconventional steps, with modified Kcx playing a key role in facilitating these processes, as revealed by molecular dynamics simulations and quantum chemical calculations. Phylogenetic analyses and other studies suggest that BTG13-related metalloenzymes are widespread in various organisms. Here, we highlight the significance of this new class of non-heme iron-dependent oxygenases and their potential as novel tools for practical applications in synthetic biology.
{"title":"BTG13-related metalloenzymes: Atypical non-heme iron-dependent dioxygenases with unusual coordination patterns and catalytic mechanisms","authors":"Zhiwei Deng,&nbsp;Zhenbo Yuan,&nbsp;Zhengshan Luo,&nbsp;Yijian Rao","doi":"10.1016/j.engmic.2024.100188","DOIUrl":"10.1016/j.engmic.2024.100188","url":null,"abstract":"<div><div>Owing to their diverse coordination patterns and catalytic mechanisms, non-heme iron-dependent dioxygenases catalyze a variety of biochemical reactions involved in the synthesis of numerous natural products and valuable compounds. Recently, we discovered a novel and atypical non-heme iron-dependent dioxygenase, BTG13, that features a unique coordination center consisting of four histidines and a carboxylated lysine (Kcx). This enzyme catalyzes the C–C bond cleavage of anthraquinone through two unconventional steps, with modified Kcx playing a key role in facilitating these processes, as revealed by molecular dynamics simulations and quantum chemical calculations. Phylogenetic analyses and other studies suggest that BTG13-related metalloenzymes are widespread in various organisms. Here, we highlight the significance of this new class of non-heme iron-dependent oxygenases and their potential as novel tools for practical applications in synthetic biology.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 1","pages":"Article 100188"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143099240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Complexity of antibiotic resistance and its impact on gut microbiota dynamics
Pub Date : 2024-12-30 DOI: 10.1016/j.engmic.2024.100187
H. Shayista , M.N. Nagendra Prasad , S. Niranjan Raj , Ashwini Prasad , S. Lakshmi , H.K. Ranjini , K. Manju , Ravikumara , Raghuraj Singh Chouhan , Olga Y. Khohlova , Olga V. Perianova , Syed Baker
The present review explores the influence of the gut microbiota on antibiotic resistance dynamics, particularly those associated with dysbiosis. The improper use of antibiotics can induce resistance in pathogens through various pathways, which is a topic of increasing interest within the scientific community. This review highlights the importance of microbial diversity, gut metabolism, and inflammatory responses against the dysbiosis due to the action of antibiotics. Additionally, it examines how secondary metabolites secreted by pathogens can serve as biomarkers for the early detection of antibiotic resistance. Although significant progress has been made in this field, key research gaps persist, including the need for a deeper understanding of the long-term effects of antibiotic-induced dysbiosis and the specific mechanisms driving the evolution of resistance in gut bacteria. Based on these considerations, this review systematically analyzed studies from PubMed, Web of Science, Embase, Cochrane Library, and Scopus up to July 2024. This study aimed to explore the dynamics of the interactions between gut microbiota and antibiotic resistance, specifically examining how microbial composition influences the development of resistance mechanisms. By elucidating these relationships, this review provides insights into management strategies for drug resistance and improves our understanding of microbial contributions to host health.
{"title":"Complexity of antibiotic resistance and its impact on gut microbiota dynamics","authors":"H. Shayista ,&nbsp;M.N. Nagendra Prasad ,&nbsp;S. Niranjan Raj ,&nbsp;Ashwini Prasad ,&nbsp;S. Lakshmi ,&nbsp;H.K. Ranjini ,&nbsp;K. Manju ,&nbsp;Ravikumara ,&nbsp;Raghuraj Singh Chouhan ,&nbsp;Olga Y. Khohlova ,&nbsp;Olga V. Perianova ,&nbsp;Syed Baker","doi":"10.1016/j.engmic.2024.100187","DOIUrl":"10.1016/j.engmic.2024.100187","url":null,"abstract":"<div><div>The present review explores the influence of the gut microbiota on antibiotic resistance dynamics, particularly those associated with dysbiosis. The improper use of antibiotics can induce resistance in pathogens through various pathways, which is a topic of increasing interest within the scientific community. This review highlights the importance of microbial diversity, gut metabolism, and inflammatory responses against the dysbiosis due to the action of antibiotics. Additionally, it examines how secondary metabolites secreted by pathogens can serve as biomarkers for the early detection of antibiotic resistance. Although significant progress has been made in this field, key research gaps persist, including the need for a deeper understanding of the long-term effects of antibiotic-induced dysbiosis and the specific mechanisms driving the evolution of resistance in gut bacteria. Based on these considerations, this review systematically analyzed studies from PubMed, Web of Science, Embase, Cochrane Library, and Scopus up to July 2024. This study aimed to explore the dynamics of the interactions between gut microbiota and antibiotic resistance, specifically examining how microbial composition influences the development of resistance mechanisms. By elucidating these relationships, this review provides insights into management strategies for drug resistance and improves our understanding of microbial contributions to host health.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 1","pages":"Article 100187"},"PeriodicalIF":0.0,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143099238","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Characterization of a small non-coding RNA S612 in Bacillus subtilis
Pub Date : 2024-12-09 DOI: 10.1016/j.engmic.2024.100186
Anqi Peng , Weijiao Zhang , Haibo Xiong , Luyao Zhang , Jian Cheng , Yang Wang , Zhen Kang
Small regulatory RNAs (sRNAs) are non-coding RNA molecules that fine-tune various cellular processes and respond to various environmental stimuli. In Bacillus subtilis, the regulatory mechanisms and specific targets of several sRNAs remain largely unknown. In this study, we identified and characterized S612 as a self-terminating sRNA in B. subtilis. The expression of S612 is regulated by external signals, including nutrient availability and salt concentration. Overexpression of S612 induced filamentous cells with extensive cellular elongation and complete inhibition of sporulation, indicating its potential to control cell morphology and spore formation. S612 directly targets and downregulates genes through post-transcriptional base pairing with mRNAs, including ylmD, trpE, ycxC, yycS, rapH, and amyE, some of which are involved in cell membrane integrity, cell wall synthesis, and sporulation initiation. Therefore, we propose that S612 is an important post-transcriptional regulator of cell morphology and sporulation.
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引用次数: 0
Terminal deoxynucleotidyl transferase: Properties and applications
Pub Date : 2024-11-28 DOI: 10.1016/j.engmic.2024.100179
Chengjie Zhang , Hizar Subthain , Fei Guo , Peng Fang , Shanmin Zheng , Mengzhe Shen , Xianger Jiang , Zhengquan Gao , Chunxiao Meng , Shengying Li , Lei Du
Terminal deoxynucleotidyl transferase (TdT), a unique DNA polymerase, can elongate DNA by adding deoxynucleotides to the 3′ terminal of a DNA chain in a template-independent manner. Owing to their remarkable DNA synthesis activity, TdTs have promoted the development of numerous nucleic acid-based methods, tools, and associated applications, attracting broad interest from both academia and industry. This review summarizes and discusses the recent research on TdTs, including their biochemical characteristics, enzyme engineering, and practical applications. New insights and perspectives on the future development of TdTs are provided.
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引用次数: 0
Exploring interspecific interaction variability in microbiota: A review 探索微生物群种间相互作用的变异性:综述
Pub Date : 2024-11-09 DOI: 10.1016/j.engmic.2024.100178
Zhong Yu , Zhihao Gan , Ahmed Tawfik , Fangang Meng
Interspecific interactions are an important component and a strong selective force in microbial communities. Over the past few decades, there has been a growing awareness of the variability in microbial interactions, and various studies are already unraveling the inner working dynamics in microbial communities. This has prompted scientists to develop novel techniques for characterizing the varying interspecific interactions among microbes. Here, we review the precise definitions of pairwise and high-order interactions, summarize the key concepts related to interaction variability, and discuss the strengths and weaknesses of emerging characterization techniques. Specifically, we found that most methods can accurately predict or provide direct information about microbial pairwise interactions. However, some of these methods inevitably mask the underlying high-order interactions in the microbial community. Making reasonable assumptions and choosing a characterization method to explore varying microbial interactions should allow us to better understand and engineer dynamic microbial systems.
种间相互作用是微生物群落的一个重要组成部分,也是一种强大的选择性力量。在过去的几十年里,人们越来越意识到微生物相互作用的可变性,各种研究已经揭示了微生物群落的内部工作动态。这促使科学家们开发新的技术来描述微生物之间不同的种间相互作用。在此,我们回顾了配对相互作用和高阶相互作用的精确定义,总结了与相互作用变异性相关的关键概念,并讨论了新兴表征技术的优缺点。具体来说,我们发现大多数方法都能准确预测或提供微生物成对相互作用的直接信息。然而,其中一些方法不可避免地会掩盖微生物群落中潜在的高阶相互作用。做出合理的假设并选择一种表征方法来探索不同的微生物相互作用,应该能让我们更好地理解和设计动态微生物系统。
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引用次数: 0
Proactive monitoring of changes in the microbial community structure in wastewater treatment bioreactors using phospholipid fatty acid analysis 利用磷脂脂肪酸分析主动监测废水处理生物反应器中微生物群落结构的变化
Pub Date : 2024-11-03 DOI: 10.1016/j.engmic.2024.100177
Lawson Mensah , Elise Cartmell , Mandy Fletton , Mark Scrimshaw , Pablo Campo
Diverse microbial community structures (MCS) in wastewater treatment plants (WWTPs) are vital for effectively removing nutrients and chemicals from wastewater. However, the regular monitoring of MCS in WWTP bioreactors remains unattractive owing to the skill and cost required for deploying modern microbial molecular techniques in the routine assessment of engineered systems. In contrast, low-resolution methods for assessing broad changes in the MCS, such as phospholipid fatty acid (PLFA) analysis, have been used effectively in soil studies for decades. Despite using PLFA analysis in soil remediation studies to capture the long-term effects of environmental changes on MCS, its application in WWTPs, where the microbial mass is dynamic and operational conditions are more fluid, remains limited. In this study, microbial communities in a controlled pilot plant and 12 full-scale activated sludge plants (ASPs) were surveyed over a two-year period using PLFA analysis. This study revealed that changes in the MCS in wastewater bioreactors could be detected using PLFA analysis. The MCS comprised 59 % Gram-negative and 9 % Gram-positive bacteria, 31 % fungi, and 1 % actinomycetes. The abundances of Gram-negative bacteria and fungi were strongly inversely correlated, with an R2 = 0.93, while the fatty acids cy17:0 and 16:1ω7c positively correlated (R2 = 0.869). Variations in temperature, solid retention time, and WWTP configuration significantly influenced the MCS in activated sludge reactors. This study showed that WWTP bioreactors can be routinely monitored using PLFA analysis, and changes in the bioreactor profile that may indicate imminent bioreactor failure can be identified.
污水处理厂(WWTP)中多样化的微生物群落结构(MCS)对于有效去除污水中的营养物质和化学物质至关重要。然而,由于在工程系统的常规评估中部署现代微生物分子技术所需的技能和成本,定期监测污水处理厂生物反应器中的微生物群落结构仍然缺乏吸引力。相比之下,低分辨率方法(如磷脂脂肪酸 (PLFA) 分析)可评估 MCS 的广泛变化,几十年来一直被有效地用于土壤研究。尽管在土壤修复研究中使用磷脂脂肪酸分析来捕捉环境变化对 MCS 的长期影响,但其在污水处理厂中的应用仍然有限,因为污水处理厂中的微生物数量是动态的,运行条件也更加多变。在这项研究中,使用 PLFA 分析法对一个受控试验工厂和 12 个正式活性污泥厂 (ASP) 的微生物群落进行了为期两年的调查。这项研究表明,使用 PLFA 分析法可以检测到废水生物反应器中微生物群落的变化。微生物控制系统包括 59 % 的革兰氏阴性菌和 9 % 的革兰氏阳性菌、31 % 的真菌和 1 % 的放线菌。革兰氏阴性菌和真菌的丰度呈强烈的反相关,R2 = 0.93,而脂肪酸 cy17:0 和 16:1ω7c 呈正相关(R2 = 0.869)。温度、固体停留时间和污水处理厂配置的变化对活性污泥反应器中的 MCS 有显著影响。这项研究表明,可以利用 PLFA 分析对污水处理厂生物反应器进行常规监测,并可识别生物反应器概况中可能预示生物反应器即将发生故障的变化。
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引用次数: 0
Immobilization of Thermomyces lanuginosus lipase on metal-organic frameworks and investigation of their catalytic properties and stability 在金属有机框架上固定热霉菌脂肪酶并研究其催化特性和稳定性
Pub Date : 2024-10-28 DOI: 10.1016/j.engmic.2024.100176
Zeynab Rangraz , Mostafa M. Amini , Zohreh Habibi
Surface adsorption is a convenient and readily available method for immobilizing enzymes on metal-organic frameworks (MOFs). Metal-organic framework-5 (MOF-5), isoreticular metal-organic frameworks-3 (IRMOF-3), and multivariate analysis of MOF-5/IRMOF-3 (MMI) with a half-amino group (-NH2) were prepared in this study. Thermomyces lanuginosus lipase (TLL) was chosen as a commercially available enzyme for immobilization on the surfaces of these MOFs. Briefly, 1.5 mg of TLL was added to 10 mg of the MOFs, and after 24 h, 67, 74, and 88% of the TLL was immobilized on MOF-5, IRMOF-3, and MMI, respectively. Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, scanning electron microscopy, energy-dispersive X-ray analysis, and Brunauer–Emmett–Teller analysis were used to characterize the resulting biocomposites. TLL@MOF-5, TLL@IRMOF-3, and TLL@MMI exhibited activities of 55, 75, and 110 U/mg, respectively. Investigation of the activity and stability of the prepared biocatalysts showed that TLL immobilized on MMI was 2.34-fold more active than free TLL. TLL@MMI exhibited high stability and activity even under harsh conditions. After 24 h of incubation in a mixture of 50% (v/v) MeOH, TLL@MMI retained 80% of its activity, whereas TLL@MOF-5 and free TLL lost 50 and 60% of their activities, respectively. TLL@MMI was used to synthesize 2-arylidenehydrazinyl-4-arylthiozole derivatives (91–98%) in a one-pot vessel by adding benzaldehydes, phenacyl bromides, and thiosemicarbazide to water. The efficiency of the 4a derivative with free TLL was 43%, whereas that with TLL@MMI was 98%.
表面吸附是将酶固定在金属有机框架(MOFs)上的一种方便易得的方法。本研究制备了金属有机框架-5(MOF-5)、等规金属有机框架-3(IRMOF-3)以及带有半氨基(-NH2)的MOF-5/IRMOF-3(MMI)的多元分析。热酵母脂肪酶(TLL)被选为固定在这些 MOF 表面的市售酶。简单地说,将 1.5 毫克 TLL 加入 10 毫克 MOF,24 小时后,分别有 67%、74% 和 88% 的 TLL 被固定在 MOF-5、IRMOF-3 和 MMI 上。傅立叶变换红外光谱、X 射线衍射、热重分析、扫描电子显微镜、能量色散 X 射线分析和布鲁瑙尔-艾美特-泰勒分析被用来表征所得到的生物复合材料。TLL@MOF-5、TLL@IRMOF-3 和 TLL@MMI 的活性分别为 55、75 和 110 U/mg 。对所制备的生物催化剂的活性和稳定性的研究表明,固定在 MMI 上的 TLL 的活性是游离 TLL 的 2.34 倍。即使在苛刻的条件下,TLL@MMI 也表现出很高的稳定性和活性。在 50%(v/v)MeOH 混合液中培养 24 小时后,TLL@MMI 保持了 80% 的活性,而 TLL@MOF-5 和游离 TLL 则分别丧失了 50% 和 60% 的活性。将 TLL@MMI 加入苯甲醛、苯基溴化物和硫代氨基甲酰肼水溶液中,在一锅容器中合成了 2-芳基肼基-4-芳基硫唑衍生物(91-98%)。使用游离 TLL 的 4a 衍生物的效率为 43%,而使用 TLL@MMI 的效率为 98%。
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引用次数: 0
The way to uncovering and utilizing marine microbial resources 发掘和利用海洋微生物资源之路
Pub Date : 2024-09-24 DOI: 10.1016/j.engmic.2024.100175
Zhi-Feng Zhang, Meng Li
Recently, Chen et al. published their breakthrough results on a marine microbial genomic catalog and genetic potentials in bioprospecting in Nature, providing unprecedented opportunities for development and utilization of genetic resources of marine microorganisms. To highlight this article, we summarized and highlighted their breakthroughs seriatim
最近,Chen 等人在《自然》杂志上发表了他们在海洋微生物基因组目录和生物勘探中的遗传潜力方面的突破性成果,为海洋微生物遗传资源的开发和利用提供了前所未有的机遇。为了突出这篇文章,我们对他们的突破性成果进行了逐一总结和强调
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引用次数: 0
Biofuel production from lignocellulose via thermophile-based consolidated bioprocessing 通过基于嗜热菌的综合生物加工技术利用木质纤维素生产生物燃料
Pub Date : 2024-09-10 DOI: 10.1016/j.engmic.2024.100174
Yilin Le , Mengqi Zhang , Pengju Wu , Huilei Wang , Jinfeng Ni
The depletion of fossil fuels and their impact on the environment have led to efforts to develop alternative sustainable fuels. While biofuel derived from lignocellulose is considered a sustainable, renewable, and green energy source, enhancing biofuel production and achieving a cost-effective bioconversion of lignocellulose at existing bio-refineries remains a challenge. Consolidated bioprocessing (CBP) using thermophiles can simplify this operation by integrating multiple processes, such as hydrolytic enzyme production, lignocellulose degradation, biofuel fermentation, and product distillation. This paper reviews recent developments in the conversion of lignocellulose to biofuel using thermophile-based CBP. First, advances in thermostable enzyme and thermophilic lignocellulolytic microorganism discovery and development for lignocellulosic biorefinery use are outlined. Then, several thermophilic CBP candidates and thermophilic microbes engineered to drive CBP of lignocellulose are reviewed. CRISPR/Cas-based genome editing tools developed for thermophiles are also highlighted. The potential applications of the Design-Build-Test-Learn (DBTL) synthetic biology strategy for designing and constructing thermophilic CBP hosts are also discussed in detail. Overall, this review illustrates how to develop highly sophisticated thermophilic CBP hosts for use in lignocellulosic biorefinery applications.
化石燃料的枯竭及其对环境的影响促使人们努力开发可持续的替代燃料。虽然从木质纤维素中提取的生物燃料被认为是一种可持续、可再生的绿色能源,但在现有的生物精炼厂中提高生物燃料产量和实现木质纤维素的生物转化的成本效益仍然是一项挑战。使用嗜热菌的综合生物处理(CBP)可将水解酶生产、木质纤维素降解、生物燃料发酵和产品蒸馏等多个过程整合在一起,从而简化操作。本文回顾了利用嗜热菌 CBP 将木质纤维素转化为生物燃料的最新进展。首先,概述了用于木质纤维素生物炼制的恒温酶和嗜热木质纤维素分解微生物的发现和开发进展。然后,综述了几种嗜热 CBP 候选者和经改造可驱动木质纤维素 CBP 的嗜热微生物。此外,还重点介绍了为嗜热微生物开发的基于 CRISPR/Cas 的基因组编辑工具。还详细讨论了设计-构建-测试-学习(DBTL)合成生物学策略在设计和构建嗜热 CBP 宿主方面的潜在应用。总之,本综述说明了如何开发高度复杂的嗜热 CBP 宿主,用于木质纤维素生物精炼应用。
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引用次数: 0
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Engineering Microbiology
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