Abstract:
:The human antimicrobial peptide LL-37 is a cationic peptide with antimicrobial activity against both Gram-positive and Gram-negative microorganisms. This work describes the development of an expression system based on Escherichia coli capable of high production of the recombinant LL-37. The fusion protein Trx-LL-37 was expressed under control of T7 promoter. The expression of T7 polymerase in the E. coli strain constructed in this work was controlled by regulation mechanisms of the arabinose promoter. The expression plasmid was stabilized by the presence of parB locus which ensured higher homology of the culture during cultivation without antibiotic selection pressure. This system was capable of producing up to 1 g of fusion protein per 1 l of culture. The subsequent semipreparative HPLC allowed us to isolate 40 mg of pure LL-37. LL-37 showed high antimicrobial activity against both Gram-negative and Gram-positive microorganisms. Its activity against Candida albicans was practically nonexistent. Minimal Inhibition Concentration (MIC) determined for E. coli was 1.65 microM; for Staphylococcus aureus 2.31 microM, and for Enterococcus faecalis 5.54 microM. The effects of cathelicidin on E. coli included the ability to permeabilize both cell membranes, as could be observed by the increase of beta-galactosidase activity in extracellular space in time. Physiological changes were studied by scanning electron microscopy; Gram-positive microorganisms did not show any visible changes in cell shapes while the changes observed on E. coli cells were evident. The results of this work show that the herein designed expression system is capable of producing adequate quantities of active human antimicrobial peptide LL-37.
journal_name
Appl Microbiol Biotechnoljournal_title
Applied microbiology and biotechnologyauthors
Krahulec J,Hyrsová M,Pepeliaev S,Jílková J,Cerný Z,Machálková Jdoi
10.1007/s00253-010-2736-7subject
Has Abstractpub_date
2010-09-01 00:00:00pages
167-75issue
1eissn
0175-7598issn
1432-0614journal_volume
88pub_type
杂志文章abstract::After crude protein of the marine yeast strains maintained in this laboratory was estimated by the method of Kjehldahl, we found that the G7a strain which was identified to be a strain of Cryptococcus aureus according to the routine identification and molecular methods contained high level of protein and could grow on...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-007-1210-7
更新日期:2007-12-01 00:00:00
abstract::Enzymatic production of biodiesel by transesterification of triglycerides and alcohol, catalyzed by lipases, offers an environmentally friendly and efficient alternative to the chemically catalyzed process while using low-grade feedstocks. Methanol is utilized frequently as the alcohol in the reaction due to its react...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-015-6700-4
更新日期:2015-11-01 00:00:00
abstract::NO (3) (-) is a major nitrogen source for plant nutrition, and plant cells store NO (3) (-) in their vacuoles. Here, we report that a unique compost made from marine animal resources by thermop...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-012-4004-5
更新日期:2013-02-01 00:00:00
abstract::The worldwide interest of the current era is to increase tendency towards the use of natural substances instead of synthetic ones. So, alternative and effective environment friendly sustainable technologies are highly needed. Due to a broad range of biological activities, fungi are considered as a significant source o...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-015-7231-8
更新日期:2016-02-01 00:00:00
abstract::A gene for a synthetic protein-based polymer, G-(VPGVG)119-VPGV, coding for the EG-120mer (elastomer), was cloned into a fungal expression vector to allow constitutive expression of the polymer controlled by the gpdA (glyceraldehyde-3-phosphate dehydrogenase) promoter sequence of Aspergillus nidulans. Stable transform...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530050942
更新日期:1997-04-01 00:00:00
abstract::The β-fructofuranosidase Xd-INV from the yeast Xanthophyllomyces dendrorhous is the largest microbial enzyme producing neo-fructooligosaccharides (neo-FOS) known to date. It mainly synthesizes neokestose and neonystose, oligosaccharides with potentially improved prebiotic properties. The Xd-INV gene comprises an open ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-6145-1
更新日期:2015-04-01 00:00:00
abstract::Two hypothetical proteins XP_001818628 and XP_001819091 (designated AoFaeB and AoFaeC, respectively), showing sequence identity with known type-C feruloyl esterases, have been found in the genomic sequence of Aspergillus oryzae. We cloned the putative A. oryzae feruloyl esterase-encoding genes and expressed them in Pi...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-009-1913-z
更新日期:2009-06-01 00:00:00
abstract::The physiological effects of 5-hydroxymethylfurfural (HMF) on Saccharomyces cerevisiae CBS 8066 in the presence and absence of furfural were studied. Experiments were carried out by pulse addition of HMF (2-4 g/l) as well as HMF (2 g/l) together with furfural (2 g/l) to batch cultivations of S. cerevisiae. Synthetic m...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530000328
更新日期:2000-06-01 00:00:00
abstract::Heterorhabditis bacteriophora is used in biological control of soil-borne insect pests in horticulture and turf. Mass production is carried out in monoxenic liquid cultures pre-incubated with the symbiont of the nematodes, the bacterium Photorhabdus luminescens, before nematode dauer juveniles (DJ) are inoculated. As ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-003-1519-9
更新日期:2004-06-01 00:00:00
abstract::Escherichia coli cells expressing mink (Mustela vison) growth hormone were grown in a batch fermentation process. The expression level was estimated to be 27% of the total cellular protein after 3 h of induction with 1 mM isopropyl beta-D-thiogalactoside (IPTG). If the expression of mink growth hormone (mGH) was induc...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0673-2
更新日期:2007-02-01 00:00:00
abstract::Site-saturation mutagenesis (SSM) has been used in directed evolution of proteins for a long time. As a special form of saturation mutagenesis, it involves individual randomization at a given residue with formation of all 19 amino acids. To date, the most efficient embodiment of SSM is a one-step PCR-based approach us...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-9041-2
更新日期:2018-07-01 00:00:00
abstract::Drought, high/low temperature, and salinity are abiotic stress factors accepted as the main reason for crop yield losses in a world with growing population and food price increases. Additional problems create nutrient limitations and particularly low P soil status. The problem of phosphate fertilizers, P plant nutriti...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-012-4224-8
更新日期:2012-08-01 00:00:00
abstract::Escherichia coli normally cannot utilize the β-glucoside sugar cellobiose as a carbon and energy source unless a stringent selection pressure for survival is present. The cellobiose-utilization phenotype can be conferred by mutations in the two cryptic operons, chb and asc. In this study, the cellobiose-utilization ph...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-011-3434-9
更新日期:2011-10-01 00:00:00
abstract::Enantioselective oxidation of racemic phenyl-1,2-ethanediol into (R)-(-)-mandelic acid by a newly isolated Brevibacterium lutescens CCZU12-1 was demonstrated. It was found that optically active (R)-(-)-mandelic acid (e.e.p > 99.9 %) is produced leaving the other enantiomer (S)-(+)-phenyl-1,2-ethanediol intact. Using f...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-013-4989-4
更新日期:2013-08-01 00:00:00
abstract::The gene from Xanthomonas campestris pv. phaseoli that is involved in the C5 pathway of delta-amino-levulinic acid (ALA) of Escherichia coli. Subcloning of deletion fragments from the initial 2.5-kilobase (kb) chromosomal fragment allowed the isolation of a 1.6-kb fragment that could complement the hemM mutation. Nucl...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/BF00173986
更新日期:1994-02-01 00:00:00
abstract::Fluoroaromatics are widely and--in recent years--increasingly used as agrochemicals, starting materials for chemical syntheses and especially pharmaceuticals. This originates from the special properties the carbon-fluorine bond is imposing on organic molecules. Hence, fluoro-substituted compounds more and more are con...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-015-6817-5
更新日期:2015-09-01 00:00:00
abstract::In this study, we investigated the possibility of using a modified hydantoinase process for the production of optically pure β-amino acids. Two aryl-substituted dihydropyrimidines D,L-6-phenyl-5,6-dihydrouracil (PheDU) and para-chloro-D,L-6-phenyl-5,6-dihydrouracil (pClPheDU) were synthesized. Hydrolysis of these nove...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-011-3691-7
更新日期:2012-06-01 00:00:00
abstract::The basidiomycetous fungus Pseudozyma aphidis is able to convert vegetable oils to abundant amounts of the biosurfactant mannosylerythritol lipid (MEL) with a unique product pattern of MEL-A, MEL-B, MEL-C, and MEL-D. To investigate the metabolism of MEL production, we analyzed the transcriptome of P. aphidis DSM 70725...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-6359-2
更新日期:2015-02-01 00:00:00
abstract::The cultivated and uncultivated bacterial communities of an activated sludge plant were studied. Two samples were taken and a total of 516 bacterial isolates were classified into groups using their whole-cell protein patterns. The distribution of bacteria into protein-pattern groups differed significantly between the ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051017
更新日期:1997-07-01 00:00:00
abstract::The upstream region of the cellobiohydrolase gene cbhA of Clostridium thermocellum F7 was sequenced. It was found that this region contains the previously sequenced gene celK encoding an enzyme closely related to CbhA (cellulosomal subunit S3). The presence of a putative transcription terminator in the 524-bp intergen...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051473
更新日期:1999-06-01 00:00:00
abstract::The production of biogas by anaerobic digestion (AD) of agricultural residues, organic wastes, animal excrements, municipal sludge, and energy crops has a firm place in sustainable energy production and bio-economy strategies. Focusing on the microbial community involved in biomass conversion offers the opportunity to...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-018-8976-7
更新日期:2018-06-01 00:00:00
abstract::In order to understand the intracellular responses in recombinant CHO (rCHO) cells producing antibody in serum-free medium (SFM) supplemented with optimized hydrolysates mixtures, yielding the highest specific growth rate (μ, SFM#S1) or the highest specific antibody productivity (q(Ab,) SFM#S2), differentially express...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-011-3106-9
更新日期:2011-03-01 00:00:00
abstract::Phytases are phosphohydrolases that initiate the sequential hydrolysis of phosphate from phytate, which is the main storage form of phosphorous in numerous plant seeds, especially in cereals and grains. Phytate is indigestible for most monogastric animals, such as poultry, swine, fish, and humans; therefore, microbial...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-9308-7
更新日期:2018-11-01 00:00:00
abstract::Aptamers are single-stranded nucleic acids or peptides identified from a randomized combinatorial library through specific interaction with the target of interest. Targets can be of any size, from small molecules to whole cells, attesting to the versatility of aptamers for binding a wide range of targets. Aptamers sho...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-016-7686-2
更新日期:2016-08-01 00:00:00
abstract::A highly hydrophobic and non-toxic colloidal silica nanoparticle/polyvinyl butyral sol-gel composite membrane was prepared on a platinum wire electrode. With diphtheria-toxoid (D-Ag) as a model antigen and encapsulation of diphtheria antibody (D-Ab) in the composite architecture, this membrane could be used for reagen...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0353-2
更新日期:2006-10-01 00:00:00
abstract::DNA microarrays have found widespread use as a flexible tool to investigate bacterial metabolism. Their main advantage is the comprehensive data they produce on the transcriptional response of the whole genome to an environmental or genetic stimulus. This allows the microbiologist to monitor metabolism and to define s...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-006-0584-2
更新日期:2006-11-01 00:00:00
abstract::Clean-up of vinyl chloride (VC)-contaminated groundwater could be enhanced by stimulating aerobic VC-oxidizing bacterial populations (e.g., methanotrophs) with amendments such as molecular oxygen. In addition, ethene gas injection could further stimulate a different group of aerobic ethene- and VC-oxidizing bacteria c...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-015-6771-2
更新日期:2015-11-01 00:00:00
abstract::Streptomyces avermitilis is well known as the producer of anthelmintic agent avermectins, which are widely used in agriculture, veterinary medicine, and human medicine. aveI encodes a TetR-family regulator, which is the homolog of AtrA. It was reported that deletion of aveI caused enhanced avermectin production. In th...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-019-10062-3
更新日期:2019-10-01 00:00:00
abstract::A chromosomal DNA fragment of 7.8 kb from Lactobacillus coryniformis CECT 5711 was cloned in Escherichia coli K-12 and was found to express a functional beta-galactosidase. Nucleotide sequence analysis showed that this fragment contained two partially overlapping genes, the lacL (1,881 bp) and the lacM (960 bp), that ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0510-7
更新日期:2006-12-01 00:00:00
abstract::Lactococcus lactis is the most important starter culture organism used in the dairy industry. Although L. lactis species have been awarded Qualified Presumption of Safety status by the European Food Safety Authority, and Generally Regarded as Safe status by the US Food and Drug Administration, some strains can produce...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-6130-8
更新日期:2015-01-01 00:00:00