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. Only the solutions containing either 70 mM HEPES or 0.85% (w/v) ammonium carbonate (pH 5.5) caused less than 10% cell leakage and the energy charge of the quenched cells was high, indicating rapid inactivation of the metabolism.The efficiency of extraction of intracellular metabolites from cell cultures depends on the extraction methods, and is expected to vary between micro-organisms. For L. plantarum, we have compared five different extraction methodologies based on (i) cold methanol, (ii) perchloric acid, (iii) boiling ethanol, (iv) chloroform/methanol (1:1) and (v) chloroform/water (1:1). Quantification of representative intracellular metabolites showed that the best extraction efficiencies were achieved with cold methanol, boiling ethanol and perchloric acid. CONCLUSION:The ammonium carbonate solution was selected as the most suitable quenching buffer for metabolomics studies in L. plantarum because (i) leakage is minimal, (ii) the energy charge indicates good fixation of metabolism, and (iii) all components are easily removed during freeze-drying. A modified procedure based on cold methanol extraction combined good extractability with mild extraction conditions and high enzymatic inactivation. These features make the combination of these quenching and extraction protocols very suitable for metabolomics studies with L. plantarum.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Faijes M,Mars AE,Smid EJdoi
10.1186/1475-2859-6-27subject
Has Abstractpub_date
2007-08-20 00:00:00pages
27issn
1475-2859pii
1475-2859-6-27journal_volume
6pub_type
杂志文章abstract:BACKGROUND:The Gram-positive bacterium Bacillus subtilis has been widely used as a cell factory for the production of proteins due to its generally regarded as safe (GRAS) nature and secretion capability. Of the known secretory pathways in B. subtilis, the majority of proteins are exported from the cytoplasm by Sec pat...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0469-8
更新日期:2016-04-28 00:00:00
abstract:INTRODUCTION:Chemical industries are constantly in search of an expeditious and environmentally benign method for producing chiral synthons. Ketoreductases have been used as catalysts for enantioselective conversion of desired prochiral ketones to their corresponding alcohol. We chose reported promiscuous ketoreductase...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-1036-2
更新日期:2018-12-03 00:00:00
abstract:BACKGROUND:The application of chemical dispersants as a response to marine oil spills is raising concerns related to their potential toxicity also towards microbes involved in oil biodegradation. Hence, oil spills occurring under marine environments necessitate the application of biodispersants that are highly active, ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0797-3
更新日期:2017-11-02 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::The success of recombinant protein expression seems unpredictable and even good yields of soluble proteins do not guarantee the correct folding. The search for soluble constructs can be performed by exploiting libraries and speeded up by automation, but these approaches are money and time consuming and the tags used f...
journal_title:Microbial cell factories
pub_type: 社论
doi:10.1186/1475-2859-3-12
更新日期:2004-10-07 00:00:00
abstract:BACKGROUND:Corynebacterium glutamicum was used as a metabolic engineering chassis for production of crude violacein (mixture of violacein and deoxyviolacein) due to Corynebacterium's GRAS status and advantages in tryptophan fermentation. The violacein is a commercially potential compound with various applications deriv...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0545-0
更新日期:2016-08-24 00:00:00
abstract:BACKGROUND:Bacteriocin-producing Lactic acid bacteria (LAB) have vast applications in human and animal health, as well as in food industry. The structural, immunity, regulatory, export and modification genes are required for effective bacteriocin biosynthesis. Variations in gene sequence, composition and organisation w...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0280-y
更新日期:2015-06-16 00:00:00
abstract::The first genome sequences of the important yeast protein production host Pichia pastoris have been released into the public domain this spring. In order to provide the scientific community easy and versatile access to the sequence, two web-sites have been installed as a resource for genomic sequence, gene and protein...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-8-53
更新日期:2009-10-16 00:00:00
abstract::BACKGROUND: A rapid microtiter plate based sandwich hybridization assay was developed for detection and quantification of single RNA species using magnetic beads. Following solution hybridization target RNA molecules were collected by biotin-streptavidin affinity binding and detected by fluorescence signal generated b...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-2-4
更新日期:2003-04-28 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
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:Numerous strategies have been developed for the display of heterologous proteins in the surface of live bacterial carriers, which can be used as vaccines, immune-modulators, cancer therapy or bioremediation. Bacterial biofilms have emerged as an interesting approach for the expression of proteins of interest...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0532-5
更新日期:2016-08-11 00:00:00
abstract:BACKGROUND:Recent years have seen a huge growth in the market of industrial yeasts with the need for strains affording better performance or to be used in new applications. Stress tolerance of commercial Saccharomyces cerevisiae yeasts is, without doubt, a trait that needs improving. Such trait is, however, complex, an...
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:Fumaric acid is a commercially important component of foodstuffs, pharmaceuticals and industrial materials, yet the current methods of production are unsustainable and ecologically destructive. RESULTS:In this study, the fumarate biosynthetic pathway involving reductive reactions of the tricarboxylic acid c...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-24
更新日期:2012-02-15 00:00:00
abstract:BACKGROUND:Co-expression of two distinct guide RNAs (gRNAs) has been used to facilitate the application of CRISPR/Cas9 system in fields such as large genomic deletion. The paired gRNAs are often placed adjacently in the same direction and expressed individually by two identical promoters, constituting direct repeats (D...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01321-4
更新日期:2020-03-10 00:00:00
abstract:BACKGROUND:Protoilludene is a valuable sesquiterpene and serves as a precursor for several medicinal compounds and antimicrobial chemicals. It can be synthesized by heterologous expression of protoilludene synthase in Escherichia coli with overexpression of mevalonate (MVA) or methylerythritol-phosphate (MEP) pathway, ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0409-7
更新日期:2016-01-19 00:00:00
abstract::The biotechnology industry has become a key element in modern societies. Within this industry, the production of recombinant enzymes and biopharmaceutical proteins is of major importance. The global markets for such recombinant proteins are growing rapidly and, accordingly, there is a continuous need for new productio...
journal_title:Microbial cell factories
pub_type: 社论
doi:10.1186/1475-2859-12-3
更新日期:2013-01-14 00:00:00
abstract:BACKGROUND:Recombinant antibodies can be produced in different formats and different expression systems. Single chain variable fragments (scFvs) represent an attractive alternative to full-length antibodies and they can be easily produced in bacteria or yeast. However, the scFvs exhibit monovalent antigen-binding prope...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-109
更新日期:2011-12-15 00:00:00
abstract:BACKGROUND:Fatty aldehydes are industrially relevant compounds, which also represent a common metabolic intermediate in the microbial synthesis of various oleochemicals, including alkanes, fatty alcohols and wax esters. The key enzymes in biological fatty aldehyde production are the fatty acyl-CoA/ACP reductases (FARs)...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0869-z
更新日期:2018-02-08 00:00:00
abstract:BACKGROUND:Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metaboli...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01466-2
更新日期:2020-11-23 00:00:00
abstract::Membrane remodeling and phospholipid biosynthesis are normally tightly regulated to maintain the shape and function of cells. Indeed, different physiological mechanisms ensure a precise coordination between de novo phospholipid biosynthesis and modulation of membrane morphology. Interestingly, the overproduction of ce...
journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/s12934-020-01433-x
更新日期:2020-09-04 00:00:00
abstract:BACKGROUND:The oleaginous yeast Yarrowia lipolytica is an organism of choice for the tailored production of various compounds such as biofuels or biopolymers. When properly engineered, it is capable of producing medium-chain-length polyhydroxyalkanoate (mcl-PHA), a biobased and biodegradable polymer that can be used as...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1140-y
更新日期:2019-05-31 00:00:00
abstract:BACKGROUND:Numerous challenges remain to achieve industrially competitive space-time yields for bio-oxidations. The ability to rapidly screen bioconversion reactions for characterization and optimization is of major importance in bioprocess development and biocatalyst selection; studies at conventional lab scale are ti...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0788-4
更新日期:2017-10-10 00:00:00
abstract:BACKGROUND:The solubility of recombinant proteins expressed in bacteria is often disappointingly low. Several strategies have been developed to improve the yield and one of the most common strategies is the fusion of the target protein with a suitable partner. Despite several reports on the successful use of each of th...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-4-34
更新日期:2005-12-13 00:00:00
abstract:BACKGROUND:Bioethanol is one of the most representative eco-friendly fuels developed to replace the non-renewable fossil fuels and is the most successful commercially available bio-conversion technology till date. With the availability of inexpensive carbon sources, such as cellulosic biomass, bioethanol production has...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1213-y
更新日期:2019-10-10 00:00:00
abstract:BACKGROUND:Due to their high energy density and compatible physical properties, several monoterpenes have been investigated as potential renewable transportation fuels, either as blendstocks with petroleum or as drop-in replacements for use in vehicles (both heavy and light-weight) or in aviation. Sustainable microbial...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1099-8
更新日期:2019-03-18 00:00:00
abstract:BACKGROUND:Bioethanol produced by the yeast Saccharomyces cerevisiae is currently one of the most promising alternatives to conventional transport fuels. Lignocellulosic hemicelluloses obtained after hydrothermal pretreatment are important feedstock for bioethanol production. However, hemicellulosic materials cannot be...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0145-9
更新日期:2014-10-12 00:00:00
abstract:BACKGROUND:The E. coli lac operon and its components have been studied for decades, and lac-derived systems are widely used for recombinant protein production. However, lac operon dynamics and induction behavior remain the paradigm of gene regulation. Recently, an HPLC-MS-based method to quantify IPTG in the medium and...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-58
更新日期:2012-05-09 00:00:00
abstract:BACKGROUND:Oxidized glutathione (GSSG) is the preferred form for industrial mass production of glutathione due to its high stability compared with reduced glutathione (GSH). In our previous study, over-expression of the mitochondrial thiol oxidase ERV1 gene was the most effective for high GSSG production in Saccharomyc...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0658-0
更新日期:2017-03-15 00:00:00
abstract::Recent evidence has demonstrated that cell-to-cell signaling is a fundamental activity carried out by numerous microorganisms. A number of specialized processes are reported to be regulated by density-dependent signaling molecules including antibiotic production, bioluminescence, biofilm formation, genetic competence,...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-1-5
更新日期:2002-12-20 00:00:00