共查询到18条相似文献,搜索用时 140 毫秒
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微生物细胞表面工程是近年来发展起来的,它利用细胞表面展示技术使外源蛋白固定化于细胞表面,从而生产微生物细胞表面蛋白。微生物细胞表面工程可用于细胞催化剂、细胞吸附剂、活疫苗、生物传感器的开发等。微生物细胞表面工程具有广阔的应用前景,但是国内对这一领域的研究刚起步。在介绍细胞表面工程的基础上,对微生物细胞表面工程技术进展进行了综述,展望了对该技术的发展。 相似文献
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酿酒酵母表面展示表达系统及应用 总被引:3,自引:0,他引:3
酵母细胞表面展示表达系统是一种固定化表达异源蛋白质的真核展示系统,即把异源靶蛋白基因序列与特定的载体基因序列融合后导入酵母细胞,利用酿酒酵母细胞内蛋白转运到膜表面的机制(GPI锚定)使靶蛋白定位于酵母细胞表面并进行表达。它利用细胞表面展示技术使外源蛋白固定化于细胞表面,从而生产微生物细胞表面蛋白,可应用于生物催化剂、细胞吸附剂、活疫苗、环境治理、蛋白质文库筛选、高亲和抗体、生物传感器、抗原/抗体库构建、免疫检测及亲和纯化、癌症诊断等领域。国内对这一方面研究较少,本文主要介绍了该技术的基本原理、研究现状、应用及其发展前景。 相似文献
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合成生物学和代谢工程是构建微生物细胞工厂、实现化学品绿色生物制造的重要方法,目前主要集中在微生物代谢网络的改造及调控上,很少考虑到微生物细胞特性的影响。形态工程通过改造微生物细胞形态相关蛋白,有目的地对微生物细胞形态及分裂方式进行合理调控,从而优化微生物细胞的特性,是降低生物炼制成本的一种新兴生物工程技术。文中首先介绍了与微生物细胞形态相关的各类蛋白,并重点总结了形态工程在生物基化学品合成方面的应用进展,包括调控细胞体积以提高胞内产物积累量、改善细胞通透性以促进胞外产物分泌、实现高密度发酵以降低生产成本、控制产物水解程度以提高产品性能。最后,提出了形态工程面临的主要问题并展望了其未来的发展趋势。 相似文献
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人类社会工业化导致各种有毒物质被排放到环境中造成严重的污染。除了自然降解外,传统的处理方法包括化学转化、物理吸附、离子交换和电化学方法等,但存在二次污染、能源需求高、投资成本高、再生效率低、低浓度废水处理效率低等缺点。细胞表面展示技术是一种通过表面锚定蛋白在细胞表面连接功能肽的新型、高效的生物技术。与细胞内和分泌物表达系统相比,微生物表面展示的蛋白质对有机溶剂、蛋白酶、温度和pH的变化表现出更强的稳定性。通过细胞培养就可以获得固定在细胞表面的蛋白酶,避免了蛋白质纯化、浓缩等繁琐的程序。此外,细胞表面展示技术是良好的单细胞水平突变体文库高通量筛选平台。综述细胞表面展示技术在环境生物修复方面的研究进展,重点介绍该技术的应用和未来发展前景。 相似文献
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细菌细胞表面展示技术是一项新的蛋白质应用技术,其体系由运载蛋白、靶蛋白和宿主菌三者构成,一般可将其分为革兰阴性菌展示体系和革兰阳性菌展示体系两大类。目前已证实多种具有锚定活性的运载蛋白,并用于不同靶蛋白的细胞表面展示体系。该技术现已被应用于活体重组疫苗的开发、蛋白质文库构建与筛选、生物传感器、全细胞生物催化剂、全细胞生物吸附与降解等多个研发领域。 相似文献
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展示表达是将目的蛋白基因与细胞表面结构蛋白融合,使目的蛋白表达并锚定于细胞表面的一项技术,微生物特别是细菌常用作展示表达的宿主。在大肠杆菌中,多种外膜蛋白、鞭毛蛋白、菌毛蛋白、脂蛋白等均已被用于表达和锚定外源蛋白,革兰氏阴性菌中,较明确的是蛋白A可用以细胞表面的整合,M6蛋白也被尝试用于乳酸菌的展示表达,酵母表达体系中,将目的蛋白与凝集素融合可表达细胞表面,菌体表面表达的蛋白容易被免疫系统识别,能引起强烈的免疫反应。其中沙门菌、大肠杆菌、链球菌等展示表达的致病细菌或病毒的抗原和毒素用于免疫动物均能产生高滴度的中和抗体,用作疫苗大有前途。本文对表达体系和在疫苗方面的应用进行了概述 相似文献
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《氨基酸和生物资源》2016,(2):7-11
冰晶核蛋白(ice nucleation protein,INP)是一种分泌型外膜蛋白,广泛分布于丁香假单胞菌,荧光假单胞菌和其他革兰氏阴性菌中。由于其在相对高温下(-2~-4℃)形成冰核的特性,INP最早应用于生物制冷领域。在细菌表面展示技术中,冰晶核蛋白作为运载蛋白得到广泛的应用。与其他的表面技术载体蛋白相比较,冰晶核蛋白具有稳定表达外源蛋白及展示分子量较大的外源蛋白的优点。INP细胞表面展示技术已被应用于全细胞生物催化剂、全细胞吸附剂和环境污染物降解剂等的开发,本文将简述INP表面展示技术的研究进展。 相似文献
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The yeast cell surface provides space to display functional proteins. Heterologous proteins can be covalently anchored to the yeast cell wall by fusing them with the anchoring domain of glycosylphosphatidylinositol (GPI)-anchored cell wall proteins (GPI-CWPs). In the yeast cell-surface display system, the anchorage position of the target protein in the cell wall is an important factor that maximizes the capabilities of engineered yeast cells because the yeast cell wall consists of a 100- to 200-nm-thick microfibrillar array of glucan chains. However, knowledge is limited regarding the anchorage position of GPI-attached proteins in the yeast cell wall. Here, we report a comparative study on the effect of GPI-anchoring domain–heterologous protein fusions on yeast cell wall localization. GPI-anchoring domains derived from well-characterized GPI-CWPs, namely Sed1p and Sag1p, were used for the cell-surface display of heterologous proteins in the yeast Saccharomyces cerevisiae. Immunoelectron-microscopic analysis of enhanced green fluorescent protein (eGFP)-displaying cells revealed that the anchorage position of the GPI-attached protein in the cell wall could be controlled by changing the fused anchoring domain. eGFP fused with the Sed1-anchoring domain predominantly localized to the external surface of the cell wall, whereas the anchorage position of eGFP fused with the Sag1-anchoring domain was mainly inside the cell wall. We also demonstrate the application of the anchorage position control technique to improve the cellulolytic ability of cellulase-displaying yeast. The ethanol titer during the simultaneous saccharification and fermentation of hydrothermally-processed rice straw was improved by 30% after repositioning the exo- and endo-cellulases using Sed1- and Sag1-anchor domains. This novel anchorage position control strategy will enable the efficient utilization of the cell wall space in various fields of yeast cell-surface display technology. 相似文献
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Yeast cell-surface display—applications of molecular display 总被引:11,自引:0,他引:11
In a cell-surface engineering system established using the yeast Saccharomyces cerevisiae, novel, so-called arming yeasts are constructed that are armed with biocatalysts in the form of enzymes, functional proteins, antibodies, and combinatorial protein libraries. Among the many advantages of the system, in which proteins are genetically displayed on the cell surface, are easy reproduction of the displayed biocatalysts and easy separation of product from catalyst. As proteins and peptides of various kinds can be displayed on the yeast cell surface, the system is expected to allow the preparation of tailor-made functional proteins. With its ability to express many of the functional proteins necessary for post-translational modification and in a range of different sizes, the yeast-based molecular display system appears uniquely useful among the various display systems so far developed. Capable of conferring novel additional abilities upon living cells, cell-surface engineering heralds a new era of combinatorial bioengineering in the field of biotechnology. This mini-review describes molecular display using yeast and its various applications. 相似文献
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Tamaru Y Ohtsuka M Kato K Manabe S Kuroda K Sanada M Ueda M 《Biotechnology progress》2006,22(4):949-953
The cell surface is a functional interface between the inside and the outside of the cell. Moreover, cells have systems for anchoring surface specific proteins and for confining surface proteins to particular domains on the cell surface. For use in bioindustrial processes applied to oral vaccination, we consider that cell-surface display systems must be useful and that the yeast Saccharomyces cerevisiae, the most suitable microorganism for practical purposes, is available as a host for genetic engineering because it can be subjected to many genetic manipulations. In particular, the rigid structure of the cell makes the yeast suitable for several of the applications. In this study, we describe the expression of one of the target antigens, 380R, from the red sea bream iridovirus (RSIV), which is one of the most common viral diseases in the cultured marine fish Pagrus major in Japan, using the arming yeast system and aiming at its application for oral vaccination. We first performed the molecular cloning and expression of the 380R antigen from RSIV in Escherichia coli. The nucleotide sequence of the 380R antigen was composed of an open reading frame (ORF) of 1360 bp encoding a protein of 453 residues. To prepare a specific antibody against the 380R antigen, the recombinant protein was overexpressed and purified in E. coli. As a result of indirect immunofluorescence with the specific antibody, we could observe the expression of the 380R antigen on the surface of the yeast cells. Thus, we have successfully prepared the source of an oral vaccine using cell-surface display technology in yeast. 相似文献
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Versatile microbial surface-display for environmental remediation and biofuels production 总被引:1,自引:0,他引:1
Surface display is a powerful technique that uses natural microbial functional components to express proteins or peptides on the cell exterior. Since the reporting of the first surface-display system in the mid-1980s, a variety of new systems have been reported for yeast, Gram-positive and Gram-negative bacteria. Non-conventional display methods are emerging, eliminating the generation of genetically modified microorganisms. Cells with surface display are used as biocatalysts, biosorbents and biostimulants. Microbial cell-surface display has proven to be extremely important for numerous applications, ranging from combinatorial library screening and protein engineering to bioremediation and biofuels production. 相似文献
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Adachi T Ito J Kawata K Kaya M Ishida H Sahara H Hata Y Ogino C Fukuda H Kondo A 《Applied microbiology and biotechnology》2008,81(4):711-719
A novel cell-surface display system was constructed in Aspergillus oryzae. Each of the five genes encoding the putative cell-wall-localized protein from the A. oryzae genome was cloned and these cell-surface anchor functions were examined by fusion to the C-terminal of the green fluorescent
protein (GFP). Using the MP1 and CWP proteins as anchor proteins, GFP signals were strongly observed on the cell surface of
recombinant A. oryzae. When these proteins were used as anchor proteins for cell-surface display of β-glucosidase from A. oryzae, enzyme activity was detected on the cell surface. In particular, β-glucosidase activity of recombinant A. oryzae using MP1, a putative glycosylphosphatidylinositol (GPI) anchor protein was higher than CWP. Based on these results, it was
concluded that the MP1 protein can act as a GPI-anchor protein in A. oryzae, and the proposed cell-surface display system using MP1 allows for the display of heterogeneous and endogenous proteins. 相似文献