A review of food-grade vectors in lactic acid bacteria: from the laboratory to their application

Lactic acid bacteria (LAB) have a long history of use in fermented foods and as probiotics. Genetic manipulation of these microorganisms has great potential for new applications in food safety, as well as in the development of improved food products and in health. While genetic engineering of LAB could have a major positive impact on the food and pharmaceutical industries, progress could be prevented by legal issues related to the controversy surrounding this technology. The safe use of genetically modified LAB requires the development of food-grade cloning systems containing only the DNA from homologous hosts or generally considered as safe organisms, and not dependent antibiotic markers. The rationale for the development of cloning vectors derived from cryptic LAB plasmids is the need for new genetic engineering tools, therefore a vision from cryptic plasmids to applications in food-grade vectors for LAB plasmids is shown in this review. Replicative and integrative vectors for the construction of food-grade vectors, and the relationship between resistance mechanism and expression systems, will be treated in depth in this paper. Finally, we will discuss the limited use of these vectors, and the problems arising from their use.     

Genetic engineering as a powerful tool to improve probiotic strains

Over the last decade, there has been increasing interest in the use of probiotic microorganisms. However, certain doubts have arisen around probiotics, because of the beneficial effects of these microorganisms are not clear yet, and in many occasions those beneficial effects have not been proven. Therefore, it would be of interest if these probiotic strains were able to acquire new attributes to allow them improve and increase their beneficial characteristics. Genetic engineering can be used for human applications; for instance, the resistance to antibiotics is removed and the probiotic bacteria are modified in its own DNA. This process can be achieved by: (1) the use of food-grade vectors derived from lactic acid bacteria and/or bifidobacteria cryptic plasmids, (2) the genes integration or deletion in the chromosome of the probiotic strain, by site-specific recombination using the attP/integrase system, or by homologous recombination, using either suicide vectors, (3) using the clustered regularly interspaced short palindromic repeats (CRISPR) and the CRISPR-associated (Cas) nuclease. Through genetic engineering, the knowledge of probiotic strains as well as its use could be improved, and the doubts about probiotics could be crumped.     

益生菌在肿瘤防治中的机制和应用

益生菌是指一类通过添加到食品或药品中,能够起到调节肠道微生态平衡,从而对人或动物产生有利影响的微生物。近年来,人们对益生菌的特性、分类、分布与营养的研究很多,特别是益生菌在抗肿瘤方面的作用引起了国内外学者的广泛关注。本文综述了益生菌应用于肿瘤预防、治疗及预后的研究情况,并对益生菌未来的研究方向提出个人的观点和展望。     

益生菌的安全性

益生菌是一类重要的微生物调节剂,对人类的营养和健康具有重大的意义。目前,益生菌具有医疗和保健作用毋庸质疑,但是对于益生菌的安全性还存在争议。本文对益生菌的安全性和不安全性研究进行综述,以利于临床正确认识、了解和使用益生菌制剂,对进一步开发利用益生菌也有积极意义。 更多还原     

益生菌和致病菌对肠道的黏附及其相互作用研究进展

探讨益生菌及致病菌对肠道上皮细胞的黏附机制,比较其黏附后的生物学效应差异,重点对二者之间的竞争性黏附作用进行了概述.在上述基础上对益生菌抑制致病菌的研究前景和发展趋势进行了展望.     

Current and future uses of real-time polymerase chain reaction and microarrays in the study of intestinal microbiota, and probiotic use and effectiveness

Probiotics are defined as live microorganisms that confer a health benefit to the host when administered in adequate amounts. In addition to human health benefits, probiotics can improve various aspects of growth and performance in livestock and poultry, as well as control undesirable microorganisms in food animals. Studies indicate that probiotics can prevent or treat certain conditions, including atopic disease in infants, food allergy, infection after surgery, acute diarrhea, and symptoms associated with irritable bowel syndrome. Understanding the complete mechanism, effectiveness, and potential use of probiotics is limited by the availability and sensitivity of current methods (i.e., culturing techniques). In recent years, real-time polymerase chain reaction (PCR) and microarrays have become prominent and promising methods to examine quantitative changes of specific members of the microbial community and the influence of probiotics on the structure and function of human and animal intestinal ecosystems. Culture-independent studies have established that only a fraction of organisms present in feces are cultivable, therefore, results obtained by cultivation are limited. Conversely, in-depth knowledge of microbial genomes has enabled real-time PCR and microarrays to be more sensitive and has resulted in precise methods for comprehensive analysis of the complex gut microbiota. Additionally, these technologies can assess the influence of intestinal microorganisms on host metabolism, nutrient status, and disease. This paper reviews method technologies and applications of real-time PCR and microarray assays as they relate to the effect and use of probiotics on the intestinal microbiota and gastrointestinal disease.     

鱼类肠道微生物与宿主免疫系统相互作用研究进展

鱼类肠道中存在大量微生物,对于维持宿主健康具有重要作用。鱼类免疫系统能够监视并调控肠道微生物组成,维持肠道菌群稳态。同时,鱼类肠道共生微生物调节鱼类免疫系统,抑制病原微生物的过度增殖,保证宿主的健康。本文回顾了鱼类肠道微生物与宿主免疫系统相互作用的研究进展,重点介绍了宿主免疫系统识别肠道微生物、塑造肠道菌群以及益生菌对宿主免疫和肠道菌群的调控等,提出了理想的益生菌应该来自动物自身胃肠道,生产中应谨慎选用非宿主来源的益生菌,以期为推动鱼类肠道功能微生物开发和应用提供理论支撑。     

Evaluation of the Role of Probiotics As a New Strategy to Eliminate Microbial Toxins: a Review

Probiotics and Antimicrobial Proteins - Probiotics are living microorganisms that have favorable effects on human and animal health. The most usual types of microorganisms recruited as probiotics...     

益生菌的功能基因导入和基因编辑

益生菌已经在临床和食品领域应用多年,其安全性和有效性已经获得人们的认可。随着分子生物学技术的发展,采用益生菌作为载体进行基因导入或基因编辑,这些遗传改造的益生菌一部分已经作为新的药品或疫苗进入到临床应用阶段。携带功能基因的益生菌定殖于肠道进行表达和缓慢释放,这类益生菌作为活体药物获得益生菌和功能基因的双重功效,可用于治疗某些疑难病症。携带蛋白质抗原基因的益生菌定殖于肠道进行表达,可诱导肠道黏膜免疫、细胞免疫和体液免疫,这是一条更安全的口服疫苗途径。成簇的规则间隔短回文重复序列(clustered regularly interspaced short palindromic repeats, CRISPR)及其相关蛋白(CRISPR-associated protein, Cas)以其高效与便捷性推动了益生菌基因编辑的发展。这篇综述介绍了CRISPR-Cas9操作系统在益生菌方面的应用。对传统遗传操作较难的益生菌采用CRISPR-Cas9技术进行基因编辑,使其基因敲除和基因突变,基因敲入和基因调控等更为简单、高效和易操作。这些CRISPR/Cas9、CRISPRa和CRISPRi技术在...     

腹泻及其微生态防治的相关进展

益生菌可能对不同病因引起的不同类型的腹泻有预防的疗效.已经成功的通过调查各种各样的益生菌和确立益生菌性质来证明其对健康的益处和预防、治疗(或缓和)腹泻的作用.其它有益生作用的微生物、某些双歧杆菌、肠球菌以及益生菌酵母菌属的酵母菌作为单个株或复合益生菌在其药用方面的价值也得到肯定.本文主要综述了益生菌在腹泻方面的治疗作用和机制.     

分子水平上益生菌研究进展

益生菌是指能在生物体内存活 ,对生物体的健康有益的一类微生物。目前已经对益生菌的作用机理进行了深入和广泛的研究 ,获得了许多使用益生菌的经验。近些年来 ,分子生物学特别是基因工程技术的发展将益生菌的理论和应用研究推向了一个新的高度。从益生菌菌株的鉴定、菌株的遗传学修饰、益生菌功能基因组学和安全性等几个方面 ,综述了近年来国际上在分子水平上研究益生菌的技术方法、获得的主要研究成果和面临的挑战 ,并提出了益生菌研究的发展方向。     

益生菌调控幼龄畜禽消化道微生物研究进展

益生菌是一类对宿主(人类或动物)有益的活性微生物,包括细菌、真菌(如酵母)等,具有促进动物生长、提高免疫力的作用,是潜在的抗生素替代品。益生菌可能通过与动物消化道微生物互作来发挥益生作用,但具体机制仍不明确。综述了基于高通量测序技术研究益生菌调控幼龄畜禽(仔猪、雏鸡、反刍动物)消化道微生物群落组成的最新进展,并提出了未来研究方向,包括益生菌如何通过与消化道微生物互作影响其功能,益生菌对于幼龄畜禽不同健康状态下肠道微生物的影响,以及宿主因素如何影响益生菌对于幼龄畜禽消化道微生物的作用效果。     

Modulation of the microbial ecology of the human colon by probiotics, prebiotics and synbiotics to enhance human health: an overview of enabling science and potential applications

The application of probiotics and prebiotics to the manipulation of the microbial ecology of the human colon has recently seen many scientific advances. The sequencing of probiotic genomes is providing a wealth of new information on the biology of these microorganisms. In addition, we are learning more about the interactions of probiotics with human cells and with pathogenic bacteria. An alternative means of modulating the colonic microbial community is by the use of prebiotic oligosaccharides. Increasing knowledge of the metabolism of prebiotics by probiotics is allowing us to consider specifically targeting such dietary intervention tools at specific population groups and specific disease states.     

益生菌辅助防治过敏性疾病的研究进展

益生菌是一类在适当的摄入数量时会对宿主机体带来益处的活体微生物,其对于机体的免疫系统具有调节作用.研究表明,益生菌可以通过调节Th1/Th2平衡,增加宿主对相关抗原的口服耐受,从而缓解过敏性症状.本文综述了近十年来研究益生菌抗过敏作用在体外试验、动物实验和临床试验三方面的进展.     

Identification and characteristics of plasmids of the strains of erythromycin-producing Streptomyces erythreus

Presence of plasmid DNA was investigated in laboratory strains 2 and 4 (NRRL 2338) of S. erythreus, as well as in strains 1 and 3 of S. erythreus subjected to improvement with respect to erythromycin production. Families of plasmids close by their molecular weights were identified in S. erythreus strains 3 and 4 (NRRL 2338). A plasmid DNA fraction of S. erythreus strain 3 was studied with electron microscopy. It enabled to identify 5 plasmids: pSE11, pSE12, pSE13, pSE14 and pSE15 with length of 5.3, 12.4, 16.3, 29.6 and 86.9 kb respectively. Using of various procedures for isolation of extrachromosomal DNA did not provide its detection in S. erythreus strains 1 and 2. At least a part of the plasmids detected in S. erythreus strains 3 and 4 (NRRL 2338) was conjugative. 32R-Labeled plasmid DNA of S. erythreus strain 3 was subjected to hydridization according to Sauthern with total DNA of the 4 strains treated with restrictases BamHI, PstI and BgIII. The studies showed that the genome of S. erythreus strain 2 was not homologous with the probe while S. erythreus strain 1 contained one of the plasmids or its part in chromosome-integrated state. In strains 3 and 4 (NRRL 2338) of S. erythreus certain plasmid DNAs were present in both autonomous and chromosome-inserted states. 32P-Labeled gene of erythromycin resistance (ermE) was subjected to hybridization according to Southern with total DNA of the 4 strains and with DNA plasmid fraction of S. erythreus strain 3. The signal was positive only in hydridization of the probe with total DNA of S. erythreus strains 1, 3, and 4 (NRRL 2338).(ABSTRACT TRUNCATED AT 250 WORDS)     

益生菌促进胃肠道健康的机制及应用

人体胃肠道内生活着大量微生物,它们影响着宿主的健康.益生菌是一种活的微生物,对维持肠黏膜屏障功能、调节免疫功能和促进营养物质的代谢吸收等具有重要作用;对肠道菌群紊乱、功能性消化不良、肠胃炎、腹泻、便秘、肠绞痛、肠易激综合征、炎症性肠病以及幽门螺杆菌感染等胃肠道疾病具有良好的应用.本文对益生菌与胃肠道健康的影响作简要概述...     

益生菌的免疫调节作用及其相关应用研究进展

益生菌的定义来自希腊语,益生菌是指:当摄入足够的剂量时可以对宿主产生有益影响的活的微生物。目前,全球对益生菌及其相关制剂的研究日益受到重视,益生菌在食品、营养品及医药领域的研究也不断深入,益生菌的免疫调节机制主要包括黏膜屏障作用、增强单核—巨噬细胞的吞噬作用、促进sIgA生成等。此外,益生菌还应用于临床相关疾病的治疗,改善受损肝脏的功能,缓解肠易激综合征的临床症状,通过调节脂肪代谢及胰岛素敏感性预防2型糖尿病的发生,预防结直肠肿瘤细胞生长及减轻肠道炎症等。本文主要就益生菌的上述相关研究进展进行综述,为益生菌的研究及相关应用提供参考。     

益生菌的研究进展及其在食品中的应用

益生菌是一类能够促进宿主肠内微生物菌群的生态平衡,对宿主健康和/或生理功能产生有益作用的活性微生物。本文就益生菌概念的发展演变、益生菌种类及其在食品中的应用研究进展进行了综述,旨在为读者提供益生菌在食品领域研究应用的基本情况及其发展趋势。     

Construction of conjugative gene transfer system between E. coli and moderately thermophilic, extremely acidophilic Acidithiobacillus caldus MTH-04

A genetic transfer system for introducing foreign genes to biomining microorganisms is urgently needed. Thus, a conjugative gene transfer system was investigated for a moderately thermophilic, extremely acidophilic biomining bacterium, Acidithiobacillus caldus MTH-04. The broad-host-range IncP plasmids RP4 and R68.45 were transferred directly into A. caldus MTH-04 from Escherichia coli by conjugation at relatively high frequencies. Additionally the broad-host-range IncQ plasmids pJRD215, pVLT33, and pVLT35 were also transferred into A. caldus MTH-04 with the help of plasmid RP4 or strains with plasmid RP4 integrated into their chromosome, such as E. coli SM10. The Km(r) and Sm(r) selectable markers from these plasmids were successfully expressed in A. caldus MTH-04. Futhermore, the IncP and IncQ plasmids were transferred back into E. coli cells from A. caldus MTH-04, thereby confirming the initial transfer of these plasmids from E. coli to A. caldus MTH-04. All the IncP and IncQ plasmids studied were stable in A. caldus MTH-04. Consequently, this development of a conjugational system for A. caldus MTH-04 will greatly facilitate its genetic study.     

益生菌的治疗现状与前景以及 对呼吸道有益菌探索的研究进展

益生菌是调节机体微生态失衡的有效途径。肝功能异常影响肠道微生物,慢性肝衰竭、2型糖尿病、动脉粥样硬化相关心血管疾病等与肠道微生态失衡密切相关。同时肠道菌群亦受环境、遗传等复合条件影响,改变菌群组成可能导致疾病的发生发展。提倡益生菌对疾病的预防、治疗、预后,改善机体微环境,提高生命质量。近年来,益生菌、益生元、合生元三方面的研究飞速发展,对肠道益生菌研发已经取得一定成果。呼吸道作为与外界相通的腔道其优势菌群已经有相关报道,但对呼吸道益生菌的探索尚不明确,呼吸道内的优势菌是否可以制成益生菌制剂尚有待研究。