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Metabolic engineering of Escherichia coli for efficient production of l-arginine. 高效生产l-精氨酸的大肠杆菌代谢工程。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 DOI: 10.1016/bs.aambs.2022.11.002
Wang Hai-De, Liu Shuai, Wang Bing-Bing, Liu Jie, Xu Jian-Zhong, Zhang Wei-Guo

As a semi-essential amino acid, l-arginine (l-Arg) plays an important role in food, health care, and medical treatment. At present, the main method of producing l-Arg is the use of microbial fermentation. Therefore, the selection and breeding of high-efficiency microbial strains is the top priority. To continuously improve the l-Arg production performance of the strains, a series of metabolic engineering strategies have been tried to transform the strains. The production of l-Arg by metabolically engineered Corynebacterium glutamicum (C. glutamicum) reached a relatively high level. Escherichia coli (E. coli), as a strain with great potential for l-Arg production, also has a large number of research strategies aimed at screening effective E. coli for producing l-Arg. E. coli also has a number of advantages over C. glutamicum in producing l-Arg. Therefore, it is of great significance to screen out excellent and stable E. coli to produce l-Arg. Here, based on recent research results, we review the metabolic pathways of l-Arg production in E. coli, the research progress of l-Arg production in E. coli, and various regulatory strategies implemented in E. coli.

作为一种半必需氨基酸,l-精氨酸(l-Arg)在食品、保健和医疗中发挥着重要作用。目前,生产l-精氨酸的主要方法是利用微生物发酵。因此,高效微生物菌株的选育是当务之急。为了不断提高菌株生产l-Arg的性能,研究人员尝试了一系列代谢工程策略对菌株进行改造。经代谢工程改造的谷氨酸棒状杆菌(C. glutamicum)的l-精氨酸产量达到较高水平。大肠杆菌(e.c oli)作为一种极具生产l-Arg潜力的菌株,也有大量旨在筛选生产l-Arg的有效大肠杆菌的研究策略。大肠杆菌在生产l-精氨酸方面也比谷氨酸梭菌有许多优势。因此,筛选优良、稳定的生产l-精氨酸的大肠杆菌具有重要意义。本文基于近年来的研究成果,综述了大肠杆菌产生l-Arg的代谢途径、大肠杆菌产生l-Arg的研究进展以及大肠杆菌实现的各种调控策略。
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引用次数: 0
Advanced imaging techniques: microscopy. 先进的成像技术:显微镜。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 DOI: 10.1016/bs.aambs.2023.01.001
Mona Golmohammadzadeh, Danielle L Sexton, Shweta Parmar, Elitza I Tocheva

For decades, bacteria were thought of as "bags" of enzymes, lacking organelles and significant subcellular structures. This stood in sharp contrast with eukaryotes, where intracellular compartmentalization and the role of large-scale order had been known for a long time. However, the emerging field of Bacterial Cell Biology has established that bacteria are in fact highly organized, with most macromolecular components having specific subcellular locations that can change depending on the cell's physiological state (Barry & Gitai, 2011; Lenz & Søgaard-Andersen, 2011; Thanbichler & Shapiro, 2008). For example, we now know that many processes in bacteria are orchestrated by cytoskeletal proteins, which polymerize into surprisingly diverse superstructures, such as rings, sheets, and tread-milling rods (Pilhofer & Jensen, 2013). These superstructures connect individual proteins, macromolecular assemblies, and even two neighboring cells, to affect essential higher-order processes including cell division, DNA segregation, and motility. Understanding these processes requires resolving the in vivo dynamics and ultrastructure at different functional stages of the cell, at macromolecular resolution and in 3-dimensions (3D). Fluorescence light microscopy (fLM) of tagged proteins is highly valuable for investigating protein localization and dynamics, and the resolution power of transmission electron microscopy (TEM) is required to elucidate the structure of macromolecular complexes in vivo and in vitro. This chapter summarizes the most recent advances in LM and TEM approaches that have revolutionized our knowledge and understanding of the microbial world.

几十年来,细菌被认为是酶的“袋子”,缺乏细胞器和重要的亚细胞结构。这与真核生物形成鲜明对比,在真核生物中,细胞内的区隔化和大尺度秩序的作用已经知道很长时间了。然而,新兴的细菌细胞生物学领域已经确定细菌实际上是高度组织化的,大多数大分子成分具有特定的亚细胞位置,可以根据细胞的生理状态而改变(Barry & Gitai, 2011;Lenz & Søgaard-Andersen, 2011;Thanbichler & Shapiro, 2008)。例如,我们现在知道细菌的许多过程是由细胞骨架蛋白策划的,这些蛋白聚合成令人惊讶的不同上层结构,如环、片和踏面磨棒(Pilhofer & Jensen, 2013)。这些上层结构连接单个蛋白质、大分子组合,甚至两个相邻的细胞,影响基本的高阶过程,包括细胞分裂、DNA分离和运动。理解这些过程需要在细胞的不同功能阶段,以大分子分辨率和三维(3D)分辨率解决体内动力学和超微结构。标记蛋白的荧光显微镜(fLM)对于研究蛋白质的定位和动力学具有重要价值,而透射电子显微镜(TEM)的分辨率对于阐明体内和体外大分子复合物的结构是必需的。本章总结了LM和TEM方法的最新进展,这些方法彻底改变了我们对微生物世界的认识和理解。
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引用次数: 0
Rhodotorula sp. as a cell factory for production of valuable biomolecules. 红酵母是生产有价值生物分子的细胞工厂。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 DOI: 10.1016/bs.aambs.2023.04.001
Cassamo U Mussagy, Helena F Ribeiro, Jorge F B Pereira

Rhodotorula sp. are well-known for their ability to biosynthesize a diverse range of valuable biomolecules, including carotenoids, lipids, enzymes, and polysaccharides. Despite the high number of studies conducted using Rhodotorula sp. at the laboratory scale, most of these do not address all processual aspects necessary for scaling up these processes for industrial applications. This chapter explores the potential of Rhodotorula sp. as a cell factory for the production of distinct biomolecules, with a particular emphasis on exploring their use from a biorefinery perspective. Through in-depth discussions of the latest research and insights into non-conventional applications, we aim to provide a comprehensive understanding of Rhodotorula sp.'s ability to produce biofuels, bioplastics, pharmaceuticals, and other valuable biochemicals. This book chapter also examines the fundamentals and challenges associated with the optimizing upstream and downstream processing of Rhodotorula sp-based processes. We believe that through this chapter, readers with different levels of expertise will gain insights into strategies for enhancing the sustainability, efficiency, and effectiveness of producing biomolecules using Rhodotorula sp.

红酵母以其生物合成多种有价值的生物分子的能力而闻名,包括类胡萝卜素、脂质、酶和多糖。尽管在实验室规模上使用红霉菌进行了大量研究,但其中大多数都没有解决扩大这些工艺用于工业应用所需的所有工艺方面。本章探讨了红霉菌作为生产不同生物分子的细胞工厂的潜力,特别强调了从生物炼制的角度探索它们的用途。通过对最新研究的深入讨论和对非常规应用的见解,我们旨在全面了解红霉菌生产生物燃料、生物塑料、药品和其他有价值的生物化学品的能力。这本书的章节还检查了基础和挑战与优化上游和下游加工的Rhodotorula sp为基础的过程。我们相信,通过本章,具有不同专业知识水平的读者将深入了解利用红霉菌生产生物分子的可持续性,效率和有效性的策略。
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引用次数: 0
Promoters and introns as key drivers for enhanced gene expression in Saccharomyces cerevisiae. 启动子和内含子是酿酒酵母基因表达增强的关键驱动因素。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 Epub Date: 2023-07-27 DOI: 10.1016/bs.aambs.2023.07.002
Marthinus Wessel Myburgh, Kirstie Susan Schwerdtfeger, Rosemary Anne Cripwell, Willem Heber van Zyl, Marinda Viljoen-Bloom

The transcription of genes in the yeast Saccharomyces cerevisiae is governed by multiple layers of regulatory elements and proteins, cooperating to ensure optimum expression of the final protein product based on the cellular requirements. Promoters have always been regarded as the most important determinant of gene transcription, but introns also play a key role in the expression of intron-encoding genes. Some introns can enhance transcription when introduced either promoter-proximal or embedded in the open reading frame of genes. However, the outcome is seldom predictable, with some introns increasing or decreasing transcription depending on the promoter and reporter gene employed. This chapter provides an overview of the general structure and function of promoters and introns and how they may cooperate during transcription to allow intron-mediated enhancement of gene expression. Since S. cerevisiae is a suitable host for recombinant protein production on a commercial level, stronger and more controllable promoters are in high demand. Enhanced gene expression can be achieved via promoter engineering, which may include introns that increase the efficacy of recombinant expression cassettes. Different models for the role of introns in transcription are briefly discussed to show how these intervening sequences can actively interact with the transcription machinery. Furthermore, recent examples of improved protein production via the introduction of promoter-proximal introns are highlighted to showcase the potential value of intron-mediated enhancement of gene expression.

酿酒酵母中基因的转录由多层调控元件和蛋白质控制,协同作用以确保基于细胞需求的最终蛋白质产物的最佳表达。启动子一直被认为是基因转录的最重要决定因素,但内含子在内含子编码基因的表达中也起着关键作用。当在启动子附近或嵌入基因的开放阅读框中时,一些内含子可以增强转录。然而,结果很少是可预测的,一些内含子的转录增加或减少取决于所使用的启动子和报告基因。本章概述了启动子和内含子的一般结构和功能,以及它们在转录过程中如何协同作用,以实现内含子介导的基因表达增强。由于酿酒酵母是在商业水平上生产重组蛋白的合适宿主,因此对更强和更可控的启动子的需求很高。增强的基因表达可以通过启动子工程实现,该工程可能包括增加重组表达盒功效的内含子。简要讨论了内含子在转录中作用的不同模型,以显示这些干预序列如何与转录机制积极相互作用。此外,最近通过引入启动子近端内含子来提高蛋白质产量的例子被强调,以展示内含子介导的基因表达增强的潜在价值。
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引用次数: 1
BioMateriOME: to understand microbe-material interactions within sustainable, living architectures. BioMateriOME:在可持续的、有生命的建筑中理解微生物与物质的相互作用。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 DOI: 10.1016/bs.aambs.2022.11.001
Beatriz Delgado Corrales, Romy Kaiser, Paula Nerlich, Armand Agraviador, Angela Sherry

BioMateriOME evolved from a prototyping process which was informed from discussions between a team of designers, architects and microbiologists, when considering constructing with biomaterials or human cohabitation with novel living materials in the built environment. The prototype has two elements (i) BioMateriOME-Public (BMP), an interactive public materials library, and (ii) BioMateriOME-eXperimental (BMX), a replicated materials library for rigorous microbiome experimentation. The prototype was installed into the OME, a unique experimental living house, in order to (1) gain insights into society's perceptions of living materials, and (2) perform a comparative analysis of indoor surface microbiome development on novel biomaterials in contrast to conventional indoor surfaces, respectively. This review summarizes the BioMateriOME prototype and its use as a tool in combining microbiology, design, architecture and social science. The use of microbiology and biological components in the fabrication of biomaterials is provided, together with an appreciation of the microbial communities common to conventional indoor surfaces, and how these communities may change in response to the implementation of living materials in our homes. Societal perceptions of microbiomes and biomaterials, are considered within the framework of healthy architecture. Finally, features of architectural design with microbes in mind are introduced, with the possibility of codifying microbial surveillance into design and construction benchmarks, standards and regulations toward healthier buildings and their occupants.

BioMateriOME是从一个原型设计过程演变而来的,这个原型设计过程是由设计师、建筑师和微生物学家团队在考虑在建筑环境中使用生物材料或人类与新型生物材料共存时讨论得出的。该原型有两个元素(1)BioMateriOME-Public (BMP),一个交互式公共材料库;(2)BioMateriOME-eXperimental (BMX),一个用于严格微生物组实验的复制材料库。该原型被安装在一个独特的实验生活屋OME中,目的是:(1)深入了解社会对生物材料的看法,(2)分别对新型生物材料与传统室内表面的室内表面微生物群发育进行比较分析。本文综述了BioMateriOME原型及其在微生物学、设计、建筑和社会科学等领域的应用。提供了生物材料制造中微生物和生物成分的使用,以及对传统室内表面常见的微生物群落的欣赏,以及这些群落如何响应我们家中生物材料的实施而发生变化。社会对微生物组和生物材料的看法是在健康建筑的框架内考虑的。最后,介绍了考虑微生物的建筑设计的特点,以及将微生物监测纳入设计和施工基准、标准和法规的可能性,以实现更健康的建筑及其居住者。
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引用次数: 0
Fundamentals of utilizing microbes in advanced cancer therapeutics: current understanding and potential applications. 在晚期癌症治疗中利用微生物的基本原理:目前的理解和潜在的应用。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 DOI: 10.1016/bs.aambs.2023.03.001
Tashmeen Kaur, Deepika Sharma

One of the biggest health related issues in the twenty-first century is cancer. The current therapeutic platforms have not advanced enough to keep up with the number of rising cases. The traditional therapeutic approaches frequently fail to produce the desired outcomes. Therefore, developing new and more potent remedies is crucial. Recently, investigating microorganisms as potential anti-cancer treatments have garnered a lot of attention. Tumor-targeting microorganisms are more versatile at inhibiting cancer than the majority of standard therapies. Bacteria preferentially gather and thrive inside tumors, where they can trigger anti-cancer immune responses. They can be further trained to generate and distribute anticancer drugs based on clinical requirements using straightforward genetic engineering approaches. To improve clinical outcomes, therapeutic strategies utilizing live tumor-targeting bacteria can be used either alone or in combination with existing anticancer treatments. On the other hand, oncolytic viruses that target cancer cells, gene therapy via viral vectors, and viral immunotherapy are other popular areas of biotechnological investigation. Therefore, viruses serve as a unique candidate for anti-tumor therapy. This chapter describes the role of microbes, primarily bacteria and viruses in anti-cancer therapeutics. The various approaches to utilizing microbes in cancer therapy are discussed and examples of microorganisms that are now in use or that are undergoing experimental research are briefly discussed. We further point out the hurdles and the prospects of microbes-based remedies for cancer treatment.

21世纪最大的健康问题之一就是癌症。目前的治疗平台还不够先进,无法跟上不断上升的病例数量。传统的治疗方法往往不能产生预期的结果。因此,开发新的和更有效的治疗方法至关重要。最近,研究微生物作为潜在的抗癌治疗方法引起了很多关注。肿瘤靶向微生物在抑制癌症方面比大多数标准疗法更通用。细菌优先在肿瘤内聚集和繁殖,在那里它们可以触发抗癌免疫反应。他们可以进一步接受培训,根据临床需要使用直接的基因工程方法生产和分发抗癌药物。为了改善临床结果,利用活肿瘤靶向细菌的治疗策略可以单独使用,也可以与现有的抗癌治疗联合使用。另一方面,针对癌细胞的溶瘤病毒、通过病毒载体的基因治疗和病毒免疫治疗是生物技术研究的其他热门领域。因此,病毒作为抗肿瘤治疗的独特候选。本章描述微生物,主要是细菌和病毒在抗癌治疗中的作用。讨论了在癌症治疗中利用微生物的各种方法,并简要讨论了目前正在使用或正在进行实验研究的微生物的例子。我们进一步指出了以微生物为基础的癌症治疗方法的障碍和前景。
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引用次数: 0
Manipulation of fungal cell wall integrity to improve production of fungal natural products. 操纵真菌细胞壁的完整性以提高真菌天然产品的产量。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-01 Epub Date: 2023-09-02 DOI: 10.1016/bs.aambs.2023.07.005
Huiling Liu, Zhengshan Luo, Yijian Rao

Fungi, as an important industrial microorganism, play an essential role in the production of natural products (NPs) due to their advantages of utilizing cheap raw materials as substrates and strong protein secretion ability. Although many metabolic engineering strategies have been adopted to enhance the biosynthetic pathway of NPs in fungi, the fungal cell wall as a natural barrier tissue is the final and key step that affects the efficiency of NPs synthesis. To date, many important progresses have been achieved in improving the synthesis of NPs by regulating the cell wall structure of fungi. In this review, we systematically summarize and discuss various strategies for modifying the cell wall structure of fungi to improve the synthesis of NPs. At first, the cell wall structure of different types of fungi is systematically described. Then, strategies to disrupt cell wall integrity (CWI) by regulating the synthesis of cell wall polysaccharides and binding proteins are summarized, which have been applied to improve the synthesis of NPs. In addition, we also summarize the studies on the regulation of CWI-related signaling pathway and the addition of exogenous components for regulating CWI to improve the synthesis of NPs. Finally, we propose the current challenges and essential strategies to usher in an era of more extensive manipulation of fungal CWI to improve the production of fungal NPs.

真菌作为一种重要的工业微生物,具有利用廉价原料作为底物、蛋白质分泌能力强等优势,在天然产物(NPs)的生产中发挥着至关重要的作用。尽管人们采用了许多代谢工程策略来增强真菌中 NPs 的生物合成途径,但作为天然屏障组织的真菌细胞壁是影响 NPs 合成效率的最后和关键步骤。迄今为止,在通过调节真菌细胞壁结构提高 NPs 合成效率方面已取得了许多重要进展。在这篇综述中,我们系统地总结和讨论了改变真菌细胞壁结构以改善 NPs 合成的各种策略。首先,系统介绍了不同类型真菌的细胞壁结构。然后,总结了通过调节细胞壁多糖和结合蛋白的合成来破坏细胞壁完整性(CWI)的策略,这些策略已被应用于改善 NPs 的合成。此外,我们还总结了调控 CWI 相关信号通路的研究,以及添加外源成分调控 CWI 以改善 NPs 合成的研究。最后,我们提出了当前的挑战和必要的策略,以迎接更广泛地操纵真菌 CWI 以提高真菌 NPs 产量的时代。
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引用次数: 0
Microbial mercury transformations: Molecules, functions and organisms. 微生物汞转化:分子、功能和有机体。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-04-13 DOI: 10.1016/bs.aambs.2022.03.001
Ri-Qing Yu,Tamar Barkay
Mercury (Hg) methylation, methylmercury (MeHg) demethylation, and inorganic redox transformations of Hg are microbe-mediating processes that determine the fate and cycling of Hg and MeHg in many environments, and by doing so influence the health of humans and wild life. The discovery of the Hg methylation genes, hgcAB, in the last decade together with advances in high throughput and genome sequencing methods, have resulted in an expanded appreciation of the diversity of Hg methylating microbes. This review aims to describe experimentally confirmed and recently discovered hgcAB gene-carrying Hg methylating microbes; phylogenetic and taxonomic analyses are presented. In addition, the current knowledge on transformation mechanisms, the organisms that carry them out, and the impact of environmental parameters on Hg methylation, MeHg demethylation, and inorganic Hg reduction and oxidation is summarized. This knowledge provides a foundation for future action toward mitigating the impact of environmental Hg pollution.
汞(Hg)甲基化、甲基汞(MeHg)去甲基化和汞的无机氧化还原转化是微生物介导的过程,决定了汞和甲基汞在许多环境中的命运和循环,并通过这样做影响人类和野生动物的健康。汞甲基化基因hgcAB的发现,以及高通量和基因组测序方法的进步,使人们对汞甲基化微生物的多样性有了更广泛的认识。本文综述了实验证实的和新近发现的携带hgcAB基因的汞甲基化微生物;提出了系统发育和分类分析。此外,对汞的转化机制、进行转化的生物以及环境参数对汞甲基化、甲基汞去甲基化和无机汞还原氧化的影响进行了综述。这一认识为今后减轻环境汞污染影响的行动奠定了基础。
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引用次数: 6
Microbial community signatures for estimation of postmortem time intervals. 用于估计死后时间间隔的微生物群落特征。
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-03-14 DOI: 10.1016/bs.aambs.2022.02.002
Hirak Ranjan Dash,Surajit Das
The human body provides a complex ecosystem for symbiotic habitation of a huge number of microorganisms. These commensal microorganisms provide a huge benefit to the living host by acting against many deadly infections. Once the host dies, many changes in the complex ecosystem of the human body take place. The personalized microbes of a human body undergo successional change as many exogenous microbes attack the nutrient-rich cadaver after death. The succession pattern change of microbes in human cadaver allows postulating different models for estimation of Postmortem time interval (PMI). Estimation of PMI has a broad prospect from the criminal investigation point of view. Though many techniques are being used nowadays to estimate PMI, all of them have their pros and cons. With the advent of advanced molecular biological techniques, studies on the thanatomicrobiome of a human cadaver have gained pace and provide a superior alternative for conventional methods of PMI estimation. This chapter summarizes the recent advancements in the changes in signature microflora postmortem with change in human microenvironment to postulate a consensus model for estimation of PMI.
人体为大量微生物的共生提供了一个复杂的生态系统。这些共生微生物通过对抗许多致命的感染,为活着的宿主提供了巨大的好处。一旦宿主死亡,人体复杂的生态系统就会发生许多变化。由于许多外源微生物在人死后攻击营养丰富的尸体,人体的个性化微生物经历了一系列的变化。人类尸体中微生物的演替模式的变化允许假设不同的模型来估计死后时间间隔(PMI)。从刑侦的角度来看,PMI估算具有广阔的应用前景。尽管目前有许多技术被用于估算PMI,但它们都有各自的优缺点。随着先进分子生物学技术的出现,对人类尸体的死亡微生物组的研究取得了进展,并为传统的PMI估算方法提供了更好的选择。本章总结了近年来人类微环境变化对死后特征菌群变化的研究进展,提出了一个估算PMI的共识模型。
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引用次数: 2
CRISPR/Cas genome editing systems in thermophiles: Current status, associated challenges, and future perspectives. 嗜热菌CRISPR/Cas基因组编辑系统:现状、相关挑战和未来展望
2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-02-25 DOI: 10.1016/bs.aambs.2022.02.001
Yilin Le,Jianzhong Sun
Thermophiles, offering an attractive and unique platform for a broad range of applications in biofuels and environment protections, have received a significant attention and growing interest from academy and industry. However, the exploration and exploitation of thermophilic organisms have been hampered by the lack of a powerful genome manipulation tool to improve production efficiency. At current, the clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/CRISPR associated (Cas) system has been successfully exploited as a competent, simplistic, and powerful tool for genome engineering both in eukaryotes and prokaryotes. Indeed, with the significant efforts made in recent years, some thermostable Cas9 proteins have been well identified and characterized and further, some thermostable Cas9-based editing tools have been successfully established in some representative obligate thermophiles. In this regard, we reviewed the current status and its progress in CRISPR/Cas-based genome editing system towards a variety of thermophilic organisms. Despite the potentials of these progresses, multiple factors/barriers still have to be overcome and optimized for improving its editing efficiency in thermophiles. Some insights into the roles of thermostable CRISPR/Cas technologies for the metabolic engineering of thermophiles as a thermophilic microbial cell factory were also fully analyzed and discussed.
嗜热菌为生物燃料和环境保护的广泛应用提供了一个有吸引力和独特的平台,受到了学术界和工业界的极大关注和日益增长的兴趣。然而,由于缺乏强大的基因组操作工具来提高生产效率,对嗜热生物的探索和开发一直受到阻碍。目前,聚集的规则间隔短回文重复序列(CRISPR)/CRISPR相关(Cas)系统已经成功地作为一种有效的、简单的、强大的工具在真核生物和原核生物中进行基因组工程。事实上,随着近年来的巨大努力,一些耐热性Cas9蛋白已经被很好地鉴定和表征,并且一些基于耐热性Cas9的编辑工具已经成功地在一些代表性的专性嗜热细菌中建立起来。在这方面,我们综述了基于CRISPR/ cas的基因组编辑系统对多种嗜热生物的研究现状及进展。尽管这些进展具有潜力,但仍需要克服和优化多个因素/障碍,以提高其在嗜热菌中的编辑效率。对热稳定性CRISPR/Cas技术作为嗜热微生物细胞工厂在嗜热菌代谢工程中的作用进行了充分的分析和讨论。
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引用次数: 2
期刊
Advances in applied microbiology
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