Abstract:
BACKGROUND:A number of valuable candidates as tuberculosis vaccine have been reported, some of which have already entered clinical trials. The new vaccines, especially subunit vaccines, need multiple administrations in order to maintain adequate life-long immune memory: this demands for high production levels and degree of purity. RESULTS:In this study, TB10.4, Ag85B and a TB10.4-Ag85B chimeric protein (here-after referred as full)--immunodominant antigens of Mycobacterium tuberculosis--were expressed in Escherichia coli and purified to homogeneity. The rational design of expression constructs and optimization of fermentation and purification conditions allowed a marked increase in solubility and yield of the recombinant antigens. Indeed, scaling up of the process guaranteed mass production of all these three antigens (2.5-25 mg of pure protein/L cultivation broth). Quality of produced soluble proteins was evaluated both by mass spectrometry to assess the purity of final preparations, and by circular dichroism spectroscopy to ascertain the protein conformation. Immunological tests of the different protein products demonstrated that when TB10.4 was fused to Ag85B, the chimeric protein was more immunoreactive than either of the immunogenic protein alone. CONCLUSIONS:We reached the goal of purifying large quantities of soluble antigens effective in generating immunological response against M. tuberculosis by a robust, controlled, scalable and economically feasible production process.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Piubelli L,Campa M,Temporini C,Binda E,Mangione F,Amicosante M,Terreni M,Marinelli F,Pollegioni Ldoi
10.1186/1475-2859-12-115subject
Has Abstractpub_date
2013-11-19 00:00:00pages
115issn
1475-2859pii
1475-2859-12-115journal_volume
12pub_type
杂志文章abstract:BACKGROUND:L-Ornithine is a non-protein amino acid with extensive applications in medicine and the food industry. Currently, L-ornithine production is based on microbial fermentation, and few microbes are used for producing L-ornithine owing to unsatisfactory production titer. RESULTS:In this study, Corynebacterium gl...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0776-8
更新日期:2017-09-22 00:00:00
abstract:BACKGROUND:Acetic acid bacteria (AAB) are widely applied in food, bioengineering and medicine fields. However, the acid stress at low pH conditions limits acetic acid fermentation efficiency and high concentration of vinegar production with AAB. Therefore, how to enhance resistance ability of the AAB remains as the maj...
journal_title:Microbial cell factories
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更新日期:2017-06-15 00:00:00
abstract:BACKGROUND:Omega hydroxy fatty acids (ω-OHFAs) are multifunctional compounds that act as the basis for the production of various industrial products with broad commercial and pharmaceutical implications. However, the terminal oxygenation of saturated or unsaturated fatty acids for the synthesis of ω-OHFAs is intricate ...
journal_title:Microbial cell factories
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doi:10.1186/s12934-015-0228-2
更新日期:2015-04-02 00:00:00
abstract:BACKGROUND:The methylotrophic yeast Pichia pastoris is a common host for the production of recombinant proteins. However, hypermannosylation hinders the use of recombinant proteins from yeast in most biopharmaceutical applications. Glyco-engineered yeast strains produce more homogeneously glycosylated proteins, but can...
journal_title:Microbial cell factories
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abstract:BACKGROUND:Despite the progressive miniaturization of bioreactors for screening purposes, shake flasks are still widespread in biotechnological laboratories and industry as cultivation vessels. Shake flasks are often applied as the first or second step in applications such as strain screening or media optimization. Thu...
journal_title:Microbial cell factories
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doi:10.1186/s12934-016-0444-4
更新日期:2016-02-24 00:00:00
abstract:BACKGROUND:C4-dicarboxylic acids, including malic acid, fumaric acid and succinic acid, are valuable organic acids that can be produced and secreted by a number of microorganisms. Previous studies on organic acid production by Aspergillus carbonarius, which is capable of producing high amounts of citric acid from varie...
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doi:10.1186/s12934-017-0660-6
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journal_title:Microbial cell factories
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doi:10.1186/s12934-015-0357-7
更新日期:2015-10-26 00:00:00
abstract::Phytases are widely utilized in feed industry to increase the utilization of phosphorus, minerals, and amino acids for improvement of animal and human nutrition. At present, all known β-propeller phytases (BPP) have been generated from bacteria, particularly Bacillus. In this work we report for the first time a new fu...
journal_title:Microbial cell factories
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更新日期:2020-04-06 00:00:00
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journal_title:Microbial cell factories
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更新日期:2020-03-10 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2014-10-01 00:00:00
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journal_title:Microbial cell factories
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doi:10.1186/s12934-015-0206-8
更新日期:2015-02-21 00:00:00
abstract:BACKGROUND:Histamine is a key mediator of the anti-inflammatory activity conferred by the probiotic organism Lactobacillus reuteri ATCC PTA 6475 in animal models of colitis and colorectal cancer. In L. reuteri, histamine synthesis and secretion requires L-histidine decarboxylase and a L-histidine/histamine exchanger. C...
journal_title:Microbial cell factories
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更新日期:2019-12-12 00:00:00
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journal_title:Microbial cell factories
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更新日期:2020-04-13 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-44
更新日期:2013-05-04 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-86
更新日期:2010-11-13 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1119-8
更新日期:2019-04-11 00:00:00
abstract:BACKGROUND:For the cultivation of Escherichia coli in bioreactors trace element solutions are generally designed for optimal growth under aerobic conditions. They do normally not contain selenium and nickel. Molybdenum is only contained in few of them. These elements are part of the formate hydrogen lyase (FHL) complex...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-7-26
更新日期:2008-08-07 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-56
更新日期:2010-07-13 00:00:00
abstract:BACKGROUND:Messenger RNA decay is an important mechanism for controlling gene expression in all organisms. The rate of the mRNA degradation directly affects the steady state concentration of mRNAs and therefore influences the protein synthesis. RNaseE has a key importance for the general mRNA decay in E. coli. While RN...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-5-38
更新日期:2006-12-12 00:00:00
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journal_title:Microbial cell factories
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更新日期:2020-07-20 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01419-9
更新日期:2020-08-12 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2020-12-10 00:00:00
abstract:BACKGROUND:To reduce the production cost of bioethanol obtained from fermentation of the sugars provided by degradation of lignocellulosic biomass (i.e., second generation bioethanol), it is necessary to screen for new enzymes endowed with more efficient biomass degrading properties. This demands the set-up of high-thr...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-58
更新日期:2010-07-16 00:00:00
abstract:BACKGROUND:Production of heterologous proteins in the E. coli periplasm, or into the extracellular fluid has many advantages; therefore naturally occurring signal peptides are selected for proteins translocation. The aim of this study was the production in high yields of a recombinant pectin lyase that is efficiently s...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-4-35
更新日期:2005-12-14 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-67
更新日期:2011-08-11 00:00:00
abstract:BACKGROUND:Nicotinamide adenine dinucleotide phosphate (NADPH) is an important cofactor ensuring intracellular redox balance, anabolism and cell growth in all living systems. Our recent multi-omics analyses of glucoamylase (GlaA) biosynthesis in the filamentous fungal cell factory Aspergillus niger indicated that low a...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01450-w
更新日期:2020-10-23 00:00:00
abstract:BACKGROUND:A key advantage of recombinant antibody technology is the ability to optimize and tailor reagents. Single domain antibodies (sdAbs), the recombinantly produced variable domains derived from camelid and shark heavy chain antibodies, provide advantages of stability and solubility and can be further engineered ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0837-z
更新日期:2017-12-12 00:00:00
abstract:BACKGROUND:In the process of ABO-incompatible (ABOi) organ transplantation, removal of anti-A and/or B antibodies from blood plasma is a promising method to overcome hyperacute rejection and allograft loss caused by the immune response between anti-A and/or B antibodies and the A and/or B antigens in the recipient. Alt...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0538-z
更新日期:2016-08-11 00:00:00
abstract:BACKGROUND:The expression of heterologous proteins in Escherichia coli is strongly affected by codon bias. This phenomenon occurs when the codon usage of the mRNA coding for the foreign protein differs from that of the bacterium. The ribosome pauses upon encountering a rare codon and may detach from the mRNA, thereby t...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-8-41
更新日期:2009-07-24 00:00:00
abstract:BACKGROUND:A reliable quenching and metabolite extraction method has been developed for Lactobacillus plantarum. The energy charge value was used as a critical indicator for fixation of metabolism. RESULTS:Four different aqueous quenching solutions, all containing 60% of methanol, were compared for their efficiency. O...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-27
更新日期:2007-08-20 00:00:00