合成生物制造2022

本文对2022年《生物工程学报》发表的与合成生物制造相关的综述和研究论文进行了评述,重点讨论了DNA测序、DNA合成、DNA编辑、基因表达调控和数学细胞模型等底层技术,酶的设计、改造和应用技术,化学品生物催化、氨基酸及其衍生物、有机酸、天然化合物、抗生素与活性肽、功能多糖、功能蛋白质等重要产品的生物制造技术,一碳化合物和生物质原料利用技术以及合成微生物组技术,以帮助读者从一个侧面了解合成生物制造相关技术和产业的发展情况。     

抗体分子结构改造与基因工程抗体

近十年来,由于免疫学技术的发展并与分子生物学、生物物理学、基因工程及电子计算机等知识技术的渗透和融合,使科学家能从分子水平对抗体分子(Ab)——即免疫球蛋白(Ig)分子的蛋白结构、空间构型、编码基因及各功能区的结构与功能特点等进行较深入的研究,从而开始了按人们的要求对Ab分子作结构的改造,设计研制具有特定反应性及功能性的Ab—Ig分子的研究领域,形成了规代免疫学与蛋白质工程结合的高技术研究——抗体基因工程。这样把70年代中期以单克隆抗体——杂交瘤技术为重要标志的现代免疫学变革继续深入进行,为免疫学的基础研究及抗体的应用研究开拓了新的方向。     

合成生物学在生物基化学品上的应用趋势及展望

合成生物学(synthetic biology)将工程学和生物学相结合, 它不同于对自然基因模拟的基因工程和对代谢途径模拟的代谢工程,而是在以基因组解析技术和化学合成技术为核心的现代生物技术基础上,以系统生物学思想和知识为指导,综合生物化学、生物物理和生物信息技术,建立基于基因和基因组、蛋白质和蛋白质组的基本要素(模块)及其组合的工程化的资源库和技术平台,旨在设计、改造、重建或制造生物分子、生物部件、生物系统、代谢途径与发育分化过程,以及具有生命活动能力的细胞和生物个体.     

全球生物制药领域研发态势分析

<正>1引言生物制药(即生物治疗药物)指运用重组DNA技术制造或提纯于生物来源的一类用于疾病预防、治疗和诊断的制品,主要包括大分子物质(如蛋白质和核酸)、疫苗和经基因工程改造后的细胞,具体包括疫苗、治疗性抗体、重组蛋白(包括融合蛋白)、基因治疗药物、细胞治疗药物、抗菌肽、细胞因子、蛋白类激素和酶等。因其具备药理活性     

新基因起源：从自然进化到人工设计

生命体系历经40多亿年的自然进化,创造了无数丰富多彩的功能基因,保障了生命体系的传承与繁荣。然而生命体系的自然进化历程极其缓慢,新的功能基因产生需要数百万年时间,无法满足快速发展的工业生产需求。利用合成生物学技术,研究人员可以依据已知的酶催化机理和蛋白质结构进行全新的基因设计与合成,按照工业生产需求快速创造全新的蛋白质催化剂,实现各种自然界生物无法催化的生物化学反应。尽管新基因设计技术展现了激动人心的应用前景,但是目前该技术还存在设计成功率不高、酶催化活性较低、合成成本较高等科技挑战。未来随着合成生物学技术的快速发展,设计、改造、合成和筛选等技术将融合为一体,为新基因设计与创建带来全新的发展机遇。     

新基因起源:从自然进化到人工设计北大核心CSCD

生命体系历经40多亿年的自然进化,创造了无数丰富多彩的功能基因,保障了生命体系的传承与繁荣。然而生命体系的自然进化历程极其缓慢,新的功能基因产生需要数百万年时间,无法满足快速发展的工业生产需求。利用合成生物学技术,研究人员可以依据已知的酶催化机理和蛋白质结构进行全新的基因设计与合成,按照工业生产需求快速创造全新的蛋白质催化剂,实现各种自然界生物无法催化的生物化学反应。尽管新基因设计技术展现了激动人心的应用前景,但是目前该技术还存在设计成功率不高、酶催化活性较低、合成成本较高等科技挑战。未来随着合成生物学技术的快速发展,设计、改造、合成和筛选等技术将融合为一体,为新基因设计与创建带来全新的发展机遇。     

合成基因制造蛋白质

英国帝国化学工业公司和莱斯特大学的科学家们业已制造了至今最大的合成基因。当他们把新基因插入细菌时,这种基因能指导细胞制造一种可能为蛋白质的生物分子。八位科学家经过一年多的努力获得的这项成就对设计更大的、能指导微生物严密模仿人体自然物质生产新药剂的基因的可能性更大了。     

蛋白质工程及其在酶学研究中的应用

蛋白质工程是生物技术中正在开发的一个新领域。这是一门从改变基因入手,定做新的蛋白质的技术。一、生物工程的新领域长期以来,人们一直希望能制造出比天然蛋白质性能更为优越的新的蛋白质,现在这一希望正在变为现实。怎样制造定做的蛋白质呢,采用经典的多肽合成方法从头合成蛋白质,虽然有几个成功的例子,然而作为一种普遍的方法,它有很多局限性。对于大的蛋白质更是力难胜任。而且合成的蛋白质并不一定折迭成它们天然的构象。因此解决合成新蛋白质的合     

生物饲料产业发展态势分析

<正>与传统饲料相比,生物饲料具有诸多优点,且目前已在畜牧业中广泛运用,具有巨大的市场潜力。文章从产品入手,结合相关专利,分析了生物饲料技术的发展趋势,并对国内外生物饲料产业的发展态势进行了总结。生物饲料这一概念是近十几年被提出的,一般是指以饲料和饲料添加剂为对象’以基因工程、蛋白质工程、发酵     

体外展示技术及其在抗体工程中的应用

体外展示技术包括核糖体展示技术、mRNA展示技术、DNA展示技术,是在无细胞蛋白质表达体系内将基因型和表型通过一定的方法连接在一起,体外高通量的筛选多肽和蛋白质的技术。抗体的产生是一个不断选择的过程,利用体外展示技术在体外选择针对某一抗原的抗体分子,并结合基因工程技术对抗体进行改造,以产生高亲和力、高特异性的抗体。体外展示技术的研究和应用已越来越广泛,有望成为下一代的抗体制备技术。     

pEGFP-植酸酶基因质粒重组构建及其在COS7细胞中表达分析

采用PCR技术扩增细菌酸性植酸aPPA2基因ORF序列,其DNA分子为1 299 bp,编码432氨基酸,蛋白质分子量约为48 kD a。此植酸酶基因被克隆到pEGFP-N3表达载体的BamH1和Pst1克隆区域,重组的pEG-FP-aPPA2重组质粒经转化到哺乳类培养细胞COS7中。重组的pEGFP-N3-aPPA2在COS7细胞中正常表达并检测出高的植酸酶活性。本研究提出的pEGFP-N3-aPPA2重组质粒构建和在哺乳类COS7细胞表达体系为植酸酶生产提供了新的技术线路。     

A novel DNA-based diagnostic test for the detection of annual and intermediate ryegrass contamination in perennial ryegrass

Perennial ryegrass (Lolium perenne L.) is a preferred choice for the turf grass industry due to its ability to provide a durable turf cover. Genetic or physical contamination of annual (L. multiflorum Lam.) or intermediate (L. hybridum) ryegrass species in perennial ryegrass is one of the major problems affecting the grass seed industry. At present, seedling root fluorescence (SRF), a biochemical marker, is used for the detection of annual ryegrass contamination. Due to the unreliability of the SRF test, the seed industry is seeking an alternative, more reliable and accurate detection method. Currently, there are no DNA tests available in ryegrass for detecting contamination with annual and intermediate ryegrass types. We developed a novel quantitative polymerase chain reaction (Q-PCR)-based DNA test for the detection of annual and/or intermediate ryegrass types in perennial ryegrass. This DNA test was designed using an insertion/deletion (InDel) site in the LpVRN2_2 (Vernalization 2) gene, which is one of the several genes controlling vernalization in ryegrass. The new DNA test is more reliable, accurate and cost-effective in detecting contamination, with a high sensitivity of 0.04% in a sample size of 5,000 seeds. Use of larger sample sizes (12.5-fold higher compared to SRF test) provided additional accuracy in detecting the level of contamination. The method has produced consistent results in 68 perennial, 26 annual and 14 intermediate ryegrass lines.     

An improved strategy based on RAPD markers efficiently identified 95 peach cultivars

DNA markers have useful applications in cultivar identification. A novel analysis approach called cultivar identification diagram (CID) was developed using DNA markers in the separation of plant individuals. This new strategy is less time- and cost-consuming, has reliable results, and was constructed for fingerprinting. Ten 11-mer primers were used to amplify the genotypes; all 95 peach genotypes (from the National Peach Germplasm Repository, in Nanjing, China) were distinguished by a combination of 54 primers. The utilization of the CID among these 95 peach cultivars was also verified by the identification of three randomly chosen groups of cultivars. This identification showed some advantages including the use of fewer primers and easy separation of all cultivars by the corresponding primers marked in the right position on the CID. This peach CID could provide the information to separate any peach cultivars of these 95, which may be of help to the peach industry in China and for the utilization of DNA markers to identify other plant species.     

植物DNA条形码研究领域文献计量学及可视化分析

为全面了解植物DNA条形码研究领域的发展和最新动态,探讨中国DNA条形码发展的状态和前景,该文利用Web of Science数据库对该研究领域进行文献计量学统计,并对引用频次、研究热点和研究前沿进行了可视化分析。结果表明:(1)中国、美国、加拿大学者在该领域文献贡献率最大,中国研究机构发文量领先,但美国、加拿大科研机构论文质量较高,影响力较大。(2) 2009年是该领域研究的高峰期,该研究领域的前沿和研究热点主要集中在物种的识别和生物多样性应用、DNA条形码候选序列筛选和鉴定技术的规范化。(3)中国学者在植物DNA条形码领域研究具有领军作用和很高的影响力,国家提倡中药产业的发展也推动了我国DNA条形码蓬勃发展,但论文的质量和影响力与美国、英国、加拿大等发达国家研究还有一定差距,应加大与发达国家科研机构合作,提高研究能力,DNA条形码技术在植物的鉴定、分类和生物多样性的保护起到非常重要的作用。这表明建立一个更全面、通用的全球植物DNA条码库以及开发新的标记并采用新的测序技术是植物DNA条形码研究的未来前景。     

Synthetic biology advances and applications in the biotechnology industry: a perspective

Synthetic biology is a logical extension of what has been called recombinant DNA (rDNA) technology or genetic engineering since the 1970s. As rDNA technology has been the driver for the development of a thriving biotechnology industry today, starting with the commercialization of biosynthetic human insulin in the early 1980s, synthetic biology has the potential to take the industry to new heights in the coming years. Synthetic biology advances have been driven by dramatic cost reductions in DNA sequencing and DNA synthesis; by the development of sophisticated tools for genome editing, such as CRISPR/Cas9; and by advances in informatics, computational tools, and infrastructure to facilitate and scale analysis and design. Synthetic biology approaches have already been applied to the metabolic engineering of microorganisms for the production of industrially important chemicals and for the engineering of human cells to treat medical disorders. It also shows great promise to accelerate the discovery and development of novel secondary metabolites from microorganisms through traditional, engineered, and combinatorial biosynthesis. We anticipate that synthetic biology will continue to have broadening impacts on the biotechnology industry to address ongoing issues of human health, world food supply, renewable energy, and industrial chemicals and enzymes.     

Epitope-driven DNA vaccine design employing immunoinformatics against B-cell lymphoma: a biotech's challenge

DNA vaccination has been widely explored to develop new, alternative and efficient vaccines for cancer immunotherapy. DNA vaccines offer several benefits such as specific targeting, use of multiple genes to enhance immunity and reduced risk compared to conventional vaccines. Rapid developments in molecular biology and immunoinformatics enable rational design approaches. These technologies allow construction of DNA vaccines encoding selected tumor antigens together with molecules to direct and amplify the desired effector pathways, as well as highly targeted vaccines aimed at specific epitopes. Reliable predictions of immunogenic T cell epitope peptides are crucial for rational vaccine design and represent a key problem in immunoinformatics. Computational approaches have been developed to facilitate the process of epitope detection and show potential applications to the immunotherapeutic treatment of cancer. In this review a number of different epitope prediction methods are briefly illustrated and effective use of these resources to support experimental studies is described. Epitope-driven vaccine design employs these bioinformatics algorithms to identify potential targets of vaccines against cancer. In this paper the selection of T cell epitopes to develop epitope-based vaccines, the need for CD4(+) T cell help for improved vaccines and the assessment of vaccine performance against tumor are reviewed. We focused on two applications, namely prediction of novel T cell epitopes and epitope enhancement by sequence modification, and combined rationale design with bioinformatics for creation of new synthetic mini-genes. This review describes the development of epitope-based DNA vaccines and their antitumor effects in preclinical research against B-cell lymphoma, corroborating the usefulness of this platform as a potential tool for cancer therapy. Achievements in the field of DNA vaccines allow to overcome hurdles to clinical translation. In a scenario where the vaccine industry is rapidly changing from a mostly empirical approach to a rational design approach, these new technologies promise to discover and develop high-value vaccines, creating a new opportunity for future markets.     

土壤宏基因组学技术及其应用

传统的基于培养的研究方法只能反映土壤中少数(0.1%～10 %)微生物的信息,而大部分微生物目前还不能培养,因而这部分微生物资源尚难以被有效地开发利用.宏基因组学是分子生物学技术应用于环境微生物生态学研究而形成的一个新概念,主要技术包括土壤DNA的提取、文库的构建和目标基因克隆的筛选.它可为揭示微生物生态功能及其分子基础提供更全面的遗传信息,并已在微生物新功能基因筛选、活性物质开发和微生物多样性研究等方面取得了显著成果.本文对土壤宏基因组学技术的方法和应用作了详细介绍.     

Biotechnological approaches to breeding and cultivation of ornamental crop plants

Recent advances in plant biotechnology hold great potential for the ornamental horticulture industry. In addition to conventional methods, breeders can now introduce genetic variation into ornamentals by the application of recombinant DNA technology. This technology is particularly useful for effecting changes in phenotypic expression encoded by single genes such as corolla and foliage color and texture, stem length, scent, temporal regulation of flowering, vase life of cut flowers and resistance to stressful environments. In part, the commercial success of this technology will depend on developing reliable methods of transformation of ornamentals and on the stability of the introduced or altered genes. In addition, new and improved strategies of in vitro culture have been commercially implemented for the propagation and breeding of a wide variety of ornamental crops and will undoubtedly play a major role in the screening and propagation of chimeric plants developed by recombinant DNA technology.     

Meeting report: Synthetic DNA - Writing with the Letters of Life: January 24, 2012, Frankfurt, Germany

The one-day meeting on Synthetic DNA (January 24, 2012) organized by and held at the DECHEMA in Frankfurt attracted about 100 participants from academia and industry interested in synthesizing DNA and its applications in synthetic biology. In recent years the cost for synthetic DNA reduced from 7€/bp to 0.35€/bp which has opened up many new possibilities for molecular biology. You can purchase the gene, cDNA, oligo library or full vector specifically for a particular expression host and apply synthetic biology principles to produce or create new drugs, vaccines or any other biotechnological products. There are, however, great concerns within society to produce organisms that do not exist in nature, and the potential misuse of them. Adressing these concerns and to use a clear terminology that do not cause misunderstandings are important issues within the field, which were also discussed at this meeting.     

Attitudes on DNA ancestry tests

The DNA ancestry testing industry is more than a decade old, yet details about it remain a mystery: there remain no reliable, empirical data on the number, motivations, and attitudes of customers to date, the number of products available and their characteristics, or the industry customs and standard practices that have emerged in the absence of specific governmental regulations. Here, we provide preliminary data collected in 2009 through indirect and direct participant observation, namely blog post analysis, generalized survey analysis, and targeted survey analysis. The attitudes include the first available data on attitudes of those of individuals who have and have not had their own DNA ancestry tested as well as individuals who are members of DNA ancestry-related social networking groups. In a new and fluid landscape, the results highlight the need for empirical data to guide policy discussions and should be interpreted collectively as an invitation for additional investigation of (1) the opinions of individuals purchasing these tests, individuals obtaining these tests through research participation, and individuals not obtaining these tests; (2) the psychosocial and behavioral reactions of individuals obtaining their DNA ancestry information with attention given both to expectations prior to testing and the sociotechnical architecture of the test used; and (3) the applications of DNA ancestry information in varying contexts.