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1.
Bacterial biofilms are associated with chronic infections due to their resistance to antimicrobial agents. Staphylococcus
aureus is a versatile human pathogen and can form biofilms on human tissues and diverse medical devices. To identify novel biofilm
inhibitors of S. aureus, the supernatants from a library of 458 Actinomycetes strains were screened. The culture supernatants (1% v/v) of more than 10 Actinomycetes strains inhibited S. aureus biofilm formation by more than 80% without affecting the growth. The culture supernatants of these biofilm-reducing Actinomycetes strains contained a protease (equivalent to 0.1 μg proteinase K ml−1), which both inhibited S. aureus biofilm formation and detached pre-existing S. aureus biofilms. This study suggests that protease treatment could be a feasible tool to reduce and eradicate S. aureus biofilms. 相似文献
2.
The foodborne pathogen Bacillus cereus can form biofilms on various food contact surfaces, leading to contamination of food products. To study the mechanisms of
biofilm formation by B. cereus, a Tn5401 library was generated from strain UW101C. Eight thousand mutants were screened in EPS, a low nutrient medium. One mutant
(M124), with a disruption in codY, developed fourfold less biofilm than the wild-type, and its defective biofilm phenotype was rescued by complementation.
Addition of 0.1% casamino acids to EPS prolonged the duration of biofilms in the wild-type but not codY mutant. When decoyinine, a GTP synthesis inhibitor, was added to EPS, biofilm formation was decreased in the wild-type but
not the mutant. The codY mutant produced three times higher protease activity than the wild-type. Zymogram and SDS-PAGE data showed that production
of the protease (∼130 kDa) was repressed by CodY. Addition of proteinase K to EPS decreased biofilm formation by the wild-type.
Using a dpp-lacZ fusion reporter system, it was shown that that the B. cereus CodY can sense amino acids and GTP levels. These data suggest that by responding to amino acids and intracellular GTP levels
CodY represses production of an unknown protease and is involved in biofilm formation. 相似文献
3.
K. Murugan K. Selvanayaki Saleh Al-Sohaibani 《World journal of microbiology & biotechnology》2011,27(7):1661-1668
Respiratory tract and device associated infections caused by biofilm forming Pseudomonas aeruginosa play a primary role in the pathogenesis and prognosis of cystic fibrosis (CF) diseases. The biofilm formed by these pathogens
attributes to the antibiotic resistance and protection from host immune response. Once established, the pathogens respond
poorly to therapeutic agents. Recently medicinal plants are largely explored as potential source of bioactive agents. In this
context the present study reports the antibiofilm activity of the folkloric medicinal plant Andrographis paniculata against biofilm forming CF causative Pseudomonas aeruginosa isolated from CF sputum. P. aeruginosa was also assessed for their growth and development of the biofilm, phylogenetic relationship and antibiotic susceptibility.
Antibiogram of the strains indicated that they were resistant to more than one antibiotic. Six extracts of A. paniculata showed significant antibiofilm activity. P. aeruginosa strains, KMS P03 and KMS P05, were found to be maximally inhibited by the methanol extract to an extent of 88.6 and 87.5%
respectively. This is the first report on antibiofilm activity of A. paniculata extracts, and our results indicate scope for development of complementary medicine for biofilm associated infections. 相似文献
4.
Livingstone JR Yoshida I Tarui Y Hirooka K Yamamoto Y Tsutui N Hirasawa E 《Journal of plant research》2002,115(5):393-400
NAD-dependent aminoaldehyde dehydrogenase (AMADH, EC 1.2.1.-) from Avena shoots was purified by DEAE Sephacel, hydroxyapatite, 5′-AMP Sepharose 4B, Mono Q, and TSK-GEL column chromatographies to
homogeneity by the criterion of native PAGE. SDS–PAGE yielded a single band at a molecular mass of 55 kDa. IEF studies showed
a band with a pI value of 5.3. In contrast to AMADHs from other species, the TSK-GEL chromatography showed that Avena AMADH exists as a monomer in the native state. The purified enzyme catalyzed the oxidations of 3-aminopropionaldehyde (APAL),
4-aminobutyraldehyde (ABAL) N-(3-aminopropyl)-4-aminobutyraldehyde (APBAL), and 4-guanidinobutyraldehyde (GBAL), but not of betaine aldehyde or indoleacetaldehyde.
The K
m values for APAL, ABAL, and GBAL were 1.5×10–6, 2.2×10–6, and 1.3×10–5 M, respectively. Although N-terminal amino acid sequence of Avena AMADH could not be determined due to a modification of the amino residue, the sequence of the fragment of AMADH cleaved by
V8 protease showed greater similarity to the barley BADH than to the pea AMADH.
Electronic Publication 相似文献
5.
Antibiotic synergy against biofilm-forming <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> 总被引:1,自引:1,他引:0
Eight antibiotics (aztreonam, ceftazidim, cefoperazon, cefepim, netilmicin, amikacin, ofloxacin and ciprofloxacin) exhibited
antimicrobial activity individually and/or in combinations against 20 wild-type biofilm-forming strains of Pseudomonas aeruginosa. The strains were less susceptible in biofilm; in 10 strains antibiotic synergy was observed for the combination of aztreonam
and ciprofloxacin. Synergy was also demonstrated in the case of β-lactams and aminoglycosides, β-lactams and fluoroquinolones,
aminoglycosides and fluoroquinolones, and for monobactams and β-lactams although the strains were resistant to the individual
antibiotics. Synergism or partial synergism was found with one or more antibiotic combinations against 32.4% of isolates. 相似文献
6.
Zhengfang Zhang Yanming Sheng Keyi Jiang Zhao Wang Yuguo Zheng Qing Zhu 《Biotechnology letters》2010,32(4):513-516
A newly isolated Bacillus megaterium with epoxide hydrolase activity resolved racemic glycidyl (o, m, p)-methylphenyl ethers to give enantiopure epoxides in 84–99% enantiomeric excess and with 21–73 enantiomeric ratios. The (S)-enantiomer was obtained from rac-glycidyl (o or m)-methylphenyl ether while the (R)-epoxides was obtained from glycidyl p-methylphenyl ether. The observations are explained at the level by enzyme-substrate docking studies. 相似文献
7.
Xiangbin Xu Jufang Bian Songbai Liu Hongmiao Song Nongnong Shi Yuezhi Tao Huizhong Wang 《Molecular breeding : new strategies in plant improvement》2011,27(3):337-346
The PROMOTION OF CELL SURVIVAL 1 (PCS1) gene, encoding an aspartic protease, has an important role in determining the fate of cells in embryonic development and
reproduction processes in Arabidopsis. To explore the potential function of the PCS1 gene in generating reproductive sterility, we placed the PCS1 gene under the control of an 1,869-bp nucleotide sequence from the 3′ end of the second intron (AG-I) of Arabidopsis AGAMOUS and CaMV 35S (–60) minimal promoter [AG-I-35S (–60)::PCS1], and introduced it into tobacco. RT–PCR results demonstrated that the PCS1 gene driven by AG-I-35S (–60) chimeric promoter was expressed only in anthers and carpels in the reproductive tissues of transgenic tobacco. Compared to
wild-type plants, all AG-I-35S (–60) and AG-I-35S (–60)::PCS1 transgenic lines showed a normal phenotype throughout the vegetative growth phase. However, during the reproductive stage,
most AG-I-35S (–60)::PCS1 transgenic plant anthers displayed delayed dehiscence, failed dehiscence, petalody and hypoplasia, and the pollen grains
had different shapes and sizes with a distorted, shrunken, or collapsed morphology. Moreover, three transgenic lines, PCS1-1,
PCS1-3 and PCS1-4, showed higher sterility than wild-type and AG-I-35S (–60) transgenic plants, respectively. These results showed that the construct of AG-I-35S (–60)::PCS1 was partially effective at preventing seed set and provided a novel sterility strategy. 相似文献
8.
Talita A. Sampaio e Silva Adriana Knob Célia R. Tremacoldi Marcia R. Brochetto-Braga Eleonora Cano Carmona 《World journal of microbiology & biotechnology》2011,27(11):2491-2497
Acid proteases represent an important group of enzymes, widely used in food, beverage and pharmaceutical industries. For most
of these applications the enzymatic preparation must be at least partially purified and free of substances that could change
the characteristics of the product or the process. Fungal proteases have replaced other sources because they are easily obtained
mainly from Mucor, Rhizopus, Penicillium
and
Aspergillus species. A strain of Aspergillus
clavatus was selected by producing high level of acid protease activity. An extracellular aspartatic protease from this strain was
purified 37.2 times with 37% recovery using (NH4)2SO4 fractionation and ion-exchange chromatography. The enzyme was found to be monomeric having a molecular mass of 30.4 kDa.
The purified enzyme is an acid protease with optimum pH of 5.5 and temperature for optimum activity of 50 °C. Its high pH
stability was verified in the range of 3.5–6.5. The acid protease was strongly inhibited by Hg+2 and partially inhibited by Cu+2, Zn+2 and Mn+2. The enzyme was sensitive to denaturing agent SDS and activated by thiol-containing reducing agent dithiotreitol (DTT). The
protease activity was not influenced by iodoacetic acid, E-64 and PMSF, while it was lightly actived by EDTA and totally inhibited
by pepstatin, with a Ki of 7.8 μM, indicating that is an aspartic protease. A.
clavatus acid protease presents interesting characteristics for biotechnological process, such as cheese and flavor manufacture and
dietary supplements, in which activity and stability in acid pH are required. 相似文献
9.
Growth of Streptococcus zooepidemicus in a 10 l bioreactor with 50 g sucrose/l and 10 g casein hydrolysate/l gave 5–6 g hyaluronic acid/l after 24–28 h. Purification
of hyaluronic acid gave a recovery of 65% with the final material having an Mr of ∼4 × 106 Da with less than 0.1% protein. 相似文献
10.
11.
To investigate the biocontrol effectiveness of the antibiotic producing bacterium, Pseudomonas aureofaciens 63–28 against the phytopathogen Rhizoctonia solani AG-4 on Petri plates and in soybean roots, growth response and induction of PR-proteins were estimated after inoculation
with P. aureofaciens 63–28 (P), with R. solani AG-4 (R), or with P. aureofaciens 63–28 + R. solani AG-4 (P + R). P. aureofaciens 63–28 showed strong antifungal activity against R. solani AG-4 pathogens in Petri plates. Treatment with P. aureofaciens 63–28 alone increased the emergence rate, shoot fresh weight, shoot dry weight and root fresh weight at 7 days after inoculation,
when compared to R. solani AG-4; P + R treatment showed similar effects. Peroxidase (POD) and β-1,3-glucanase activity of P. aureofaciens 63–28 treated roots increased by 41.1 and 49.9%, respectively, compared to control roots. POD was 26% greater in P + R treated
roots than R. solani treated roots. Two POD isozymes (59 and 27 kDa) were strongly induced in P + R treated roots. The apparent molecular weight
of chitinase from treated roots, as determined through SDS-PAGE separation and comparison with standards, was about 29 kDa.
Five β-1,3-glucanase isozymes (80, 70, 50, 46 and 19 kDa) were observed in all treatments. These results suggest that inoculation
of soybean plants with P. aureofaciens 63–28 elevates plant growth inhibition by R. solani AG-4 and activates PR-proteins, potentially through induction of systemic resistance mechanisms. 相似文献
12.
V. Yu. Kryukov V. P. Khodyrev O. N. Yaroslavtseva A. S. Kamenova B. A. Duisembekov V. V. Glupov 《Applied Biochemistry and Microbiology》2009,45(5):511-516
A synchronous coinfection of the Colorado potato beetle Leptinotarsa decemlineata (Say) with the entomopathogenic bacteria Bacillus thuringiensis ssp. morrisoni Bonnifoi & de Barjak var. tenebrionis Krieg et al. and hyphomycete Metarhizium anisopliae (Metsch.) Sorokin or Beauveria bassiana (Bals.) Vuill leads to the rapid death of 95–100% of larvae. The bacteria arrest the nutrition of insects, while the fungal
spores kill the weakened larvae. The synergistic effect of two pathogens is recorded at a relatively low hyphomycete titer
(1–5 × 106 conidia/ml) and is evident in the mortality dynamics at all larval ages. These bacterial and fungal pathogens display no
antagonism on artificial nutrient media. This microbial complex is highly efficient under natural conditions (80–90% larval
mortality rate and no plant defoliation). 相似文献
13.
Humulus japonicus in communities of Miscanthus sacchariflorus and Phragmites australis can grow large enough to overtop other species in the Amsa-dong floodplain. Because of strong winds and the weight of Humulus, plants of M. sacchariflorus and P. australis fell in mid-August and were subject to decomposition under its dense shading. To assess the effects of H. japonicus on nutrient cycling in these communities, we collected fresh samples of M. sacchariflorus and P. australis in litterbags and decomposed them under H. japonicus for 9 months, beginning in August. Biomass and organic contents from M. sacchariflorus during this incubation period were 49–51% and 44–48%, whereas those of P. australis were 49–61% and 32–52%, respectively. Their annual k values were 1.61–1.74 and 1.46–3.54, respectively. Initial N concentrations in M. sacchariflorus and P. australis were 13 and 20 mg g−1, while C:N ratios were 31 and 21, respectively. These results indicate that H. japonicus is responsible for the collapse of M. sacchariflorus and P. australis in August and also accelerates their nutrient cycling through rapid decomposition, thereby increasing nutrient circulation
in floodplains. 相似文献
14.
15.
Uroz S Oger P Chhabra SR Cámara M Williams P Dessaux Y 《Archives of microbiology》2007,187(3):249-256
Abtsract
Comamonas strain D1 enzymatically inactivates quorum-sensing (QS) signal molecules of the N-acyl homoserine lactone (N-AHSL) family, and exhibits the broadest inactivation range of known bacteria. It degrades N-AHSL with acyl-side chains ranging from 4 to 16 carbons, with or without 3-oxo or 3-hydroxy substitutions. N-AHSL degradation yields HSL but not N-acyl homoserine: strain D1 therefore harbors an amidohydrolase activity. Strain D1
is the fifth bacterium species in which an N-AHSL amidohydrolase is described. Consistent with its N-AHSL degradation ability, strain D1 efficiently quenches various QS-dependent functions in other bacteria, such as violacein
production by Chromobacterium violaceum and pathogenicity and antibiotic production in Pectobacterium. 相似文献
16.
Hoshino T 《Applied microbiology and biotechnology》2011,91(6):1463-1475
Violacein is a natural violet pigment produced by several Gram-negative bacteria, including Chromobacterium violaceum, Janthinobacterium lividum, and Pseudoalteromonas tunicata D2, among others. This pigment has potential medical applications as antibacterial, anti-trypanocidal, anti-ulcerogenic,
and anticancer drugs. The structure of violacein consists of three units: a 5-hydroxyindole, an oxindole, and a 2-pyrrolidone.
The biosynthetic origins of hydrogen, nitrogen, and carbon in the pyrrolidone nucleus were established by feeding experiments
using various stable isotopically labeled tryptophans (Trps). Pro-S hydrogen of CH2 at the 3-position of Trp is retained during biosynthesis. The nitrogen atom is exclusively from the α-amino group, and the
skeletal carbon atoms originate from the side chains of the two Trp molecules. All three oxygen atoms in the violacein core
are derived from molecular oxygen. The most interesting biosynthetic mechanism is the 1,2-shift of the indole nucleus on the
left side of the violacein scaffold. The alternative Trp molecule is directly incorporated into the right side of the violacein
core. This indole shift has been observed only in violacein biosynthesis, despite the large number of natural products having
been isolated. There were remarkable advances in biosynthetic studies in 2006–2008. During the 3 years, most of the intermediates
and the complete pathway were established. Two independent processes are involved: the enzymatic process catalyzed by the
five proteins VioABCDE or the alternative nonenzymatic oxidative decarboxylation reactions. The X-ray crystallographic structure
of VioE that mediates the indole rearrangement reaction was recently identified, and the mechanism of the indole shift is
discussed here. 相似文献
17.
Genome predictions based on selected genes would be a very welcome approach for taxonomic studies. We analyzed three genes,
recN, flaA, and ftsY, for determining if these genes are useful tools for systematic analyses in the genus Anoxybacillus. The genes encoding a DNA repair and genetic recombination protein (recN), the flagellin protein (flaA), and GTPase signal docking protein (ftsY) were sequenced for ten Anoxybacillus species. The sequence comparisons revealed that recN sequence similarities range between 61% and 99% in the genus Anoxybacillus. Comparisons to other bacterial recN genes indicated that levels of similarity did not differ from the levels within genus Anoxybacillus. These data showed that recN is not a useful marker for the genus Anoxybacillus. A 550–600-bp region of the flagellin gene was amplified for all Anoxybacillus strains except for Anoxybacillus contaminans. The sequence similarity of flaA gene varies between 61% and 76%. Comparisons to other bacterial flagellin genes obtained from GenBank (Bacillus, Pectinatus, Proteus, and Vibrio) indicated that the levels of similarity were lower (3–42%). Based on these data, we concluded that the variability in this
single gene makes it a particularly useful marker. Another housekeeping gene ftsY suggested to reflect the G+C (mol/mol) content of whole genome was analyzed for Anoxybacillus strains. A mean difference of 1.4% was observed between the G+C content of the gene ftsY and the G+C content of the whole genome. These results showed that the gene ftsY can be used to represent whole G+C content of the Anoxybacillus species. 相似文献
18.
Dharmaprakash Viszwapriya Ganapathy Ashwinkumar Subramenium Solai Radhika Shunmugiah Karutha Pandian 《Antonie van Leeuwenhoek》2017,110(1):153-165
Streptococcus mutans, a multivirulent pathogen is considered the primary etiological agent in dental caries. Development of antibiotic resistance in the pathogen has created a need for novel antagonistic agents which can control the virulence of the organism and reduce resistance development. The present study demonstrates the in vitro anti-virulence potential of betulin (lup-20(29)-ene-3β,28-diol), an abundantly available plant triterpenoid against S. mutans UA159. Betulin exhibited significant dose dependent antibiofilm activity without affecting bacterial viability. At 240 µg/ml (biofilm inhibitory concentration), betulin inhibited biofilm formation and adherence to smooth glass surfaces by 93 and 71 % respectively. It reduced water insoluble glucan synthesis by 89 %, in conjunction with down regulation of gtfBC genes. Microscopic analysis confirmed the disruption in biofilm architecture and decreased exopolysaccharide production. Acidogenicity and aciduricity, key virulence factors responsible for carious lesions, were also notably affected. The induced auto-aggregation of cells upon treatment could be due to the down regulation of vicK. Results of gene expression analysis demonstrated significant down-regulation of virulence genes upon betulin treatment. Furthermore, the nontoxic effect of betulin on peripheral blood mononuclear cells even after 72 h treatment makes it a strong candidate for assessing its suitability to be used as a therapeutic agent. 相似文献
19.
Aguilar-Tipacamú G Rosario-Cruz R Miller RJ Guerrero FD Rodriguez-Vivas RI García-Vázquez Z 《Experimental & applied acarology》2011,54(3):301-311
Dialelic crosses and backcrosses of pyrethroid resistant (RR) and susceptible (SS) Rhipicephalus (Boophilus) microplus tick strains were carried out and the substitution (Phe-Ile) within the sodium channel gene was monitored in order to analyze
the effects of the genotype on the pyrethroid resistance phenotype as measured by the larval packet test (LPT). Parental strains:
susceptible (SS) and resistant (RR); dialelic crosses: RS (♂RR × ♀SS), and SR (♂SS × ♀RR); and backcrosses: RS × SS, RS × RR,
SR × SS and SR × RR were infested on 280 kg calves. Resistance type (monogenic or polygenic) and effective dominance were
determined based on the discriminant concentration (DC) for cipermethrine (0.5%), deltamethrine (0.09%) and flumethrine (0.01%).
Allele specific PCR (AS-PCR) was used for genotyping, looking at a sodium channel mutation (Phe-Ile substitution). The mortality
rates and allele frequency of susceptible and pyrethroid resistant reference strains were 0% mortality and 90% RR alleles
for resistant strain, and 100% mortality and 0% RR alleles as measured by the larval packet test (LPT) and allele specific
PCR (AS-PCR) respectively. Backcrossed strain SR × RR showed an effective dominance (DML) of 0.605 for cypermethrin, 0.639 for deltamethrin and 0.498 for flumethrin, while survival of backcrosses RS × SS, RS × RR
and SR × SS showed a significant tendency to recesivity. Backcrossed strain SR × RR (69.4%) also showed a higher RR genotype
frequency with regards to RS × SS (25.5%), RS × RR (36.7%) and SR × SS (32.0%), however, susceptible allele was inherited
in general as an incomplete dominant trait. Monogenic inheritance hypothesis was tested and the results showed monogenic inheritance
for cypermethrin and flumethrin (P < 0.05) but not for deltamethrin (P > 0.05). However, significant correlation was found between RR genotype and the survival rate for all three pyrethroids used
(P < 0.05), suggesting that a single substitution on the sodium channel gene can be responsible for resistance to pyrethroids
as a class, due to the high frequency for RR genotypes. Combination with different mutations or metabolic resistance mechanisms
cannot be excluded. 相似文献
20.
Recently, the prenyltransferase SirD was found to be responsible for the O-prenylation of tyrosine in the biosynthesis of sirodesmin PL in Leptosphaeria maculans. In this study, the behavior of SirD towards phenylalanine/tyrosine and tryptophan derivatives was investigated. Product
formation has been observed with 12 of 19 phenylalanine/tyrosine derivatives. It was shown that the alanine structure attached
to the benzene ring and an electron donor, e.g., OH or NH2, at its para-position are essential for the enzyme activity. Modifications were possible both at the side chain and the benzene ring.
Enzyme products from seven phenylalanine/tyrosine derivatives were isolated and characterized by MS and NMR analyses including
HSQC and HMBC and proven to be O- or N-prenylated derivatives at position C4 of the benzene rings. K
M
values of six selected derivatives were found in the range of 0.10–0.68 mM. Catalytic efficiencies (K
cat/K
M
) were determined in the range of 430–1,110 s−1·M−1 with l-tyrosine as the best substrate. In addition, 7 of 14 tested tryptophan analogs were also accepted by SirD and converted to
C7-prenylated derivatives, which was confirmed by comparison with products obtained from enzyme assays using a 7-dimethylallyltryptophan
synthase 7-DMATS from Aspergillus fumigatus. 相似文献