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1.
Isolation and characterization of dihydromonacolin-MV from <Emphasis Type="Italic">Monascus purpureus</Emphasis> for antioxidant properties 总被引:4,自引:0,他引:4
Dhale MA Divakar S Kumar SU Vijayalakshmi G 《Applied microbiology and biotechnology》2007,73(5):1197-1202
The methanolic extract of Monascus purpureus cultivated by solid-state fermentation on rice showed strong 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity
and better yield as compared to other polarity based extracted fractions. It was selected for further purification of the
antioxidant. The activity-guided repeated fractionation of methanolic extract on a silica gel column chromatography yielded
a compound that exhibited strong antioxidant activity. Based on the spectroscopic analysis by UV, IR, 1H NMR, 13C NMR, 2D-HSQCT NMR, and MS, the antioxidant isolated was elucidated as a derivative of dihydromonacolin-K, where the ester
group is 2-methyl propionate, designated as dihydromonacolin-MV. The DPPH radical was significantly scavenged by the dihydromonacolin-MV
(IC50 20±1 μg ml−1). The dihydromonacolin-MV showed strong inhibition of lipid peroxidation in a liposome model with an IC50 value of 5.71±0.38 μg ml−1 and superoxide radical scavenging activity with an IC50 value of 163.97±2.68 μg ml−1. 相似文献
2.
The effect of agitation and aeration on the growth and antibiotic production by Xenorhabdus nematophila YL001 grown in batch cultures were investigated. Efficiency of aeration and agitation was evaluated through the oxygen mass
transfer coefficient (K
L
a). With increase in K
L
a, the biomass and antibiotic activity increased. Activity units of antibiotic and dry cell weight were increased to 232 U ml−1 and 19.58 g l−1, respectively, productivity in cell and antibiotic was up more than 30% when K
L
a increased from 115.9 h−1 to 185.7 h−1. During the exponential growth phase, DO concentration was zero, the oxygen supply was not sufficient. So, based on process
analysis, a three-stage oxygen supply control strategy was used to improved the DO concentration above 30% by controlling
the agitation speed and aeration rate. The dry cell weight and activity units of antibiotic were further increased to 24.22 g l−1 and 249 U ml−1, and were improved by 24.0% and 7.0%, compared with fermentation at a constant agitation speed and a constant aeration rate
(300 rev min−1, 2.5 l min−1). 相似文献
3.
M. Salam N. Dahiya R. Sharma S. K. Soni G. S. Hoondal R. Tewari 《Indian journal of microbiology》2008,48(3):358-364
A chitinase producing bacterium Enterobacter sp. NRG4, previously isolated in our laboratory, has been reported to have a wide range of applications such as anti-fungal
activity, generation of fungal protoplasts and production of chitobiose and N-acetyl D-glucosamine from swollen chitin. In
this paper, the gene coding for Enterobacter chitinase has been cloned and expressed in Escherichia coli BL21(DE3). The structural portion of the chitinase gene comprised of 1686 bp. The deduced amino acid sequence of chitinase
has high degree of homology (99.0%) with chitinase from Serratia marcescens. The recombinant chitinase was purified to near homogeneity using His-Tag affinity chromatography. The purified recombinant
chitinase had a specific activity of 2041.6 U mg−1. It exhibited similar properties pH and temperature optima of 5.5 and 45°C respectively as that of native chitinase. Using
swollen chitin as a substrate, the Km, kcat and catalytic efficiency (kcat/Km) values of recombinant chitinase were found to be 1.27 mg ml−1, 0.69 s−1 and 0.54 s−1M−1 respectively. Like native chitinase, the recombinant chitinase produced medicinally important N-acetyl D-glucosamine and
chitobiose from swollen chitin and also inhibited the growth of many fungi. 相似文献
4.
Two new effective microbial producers of inulinases were isolated from Jerusalem artichoke tubers grown in Thailand and identified
as Aspergillus niger TISTR 3570 and Candida
guilliermondii TISTR 5844. The inulinases produced by both these microorganisms were appropriate for hydrolysing inulin to fructose as the
principal product. An initial inulin concentration of ∼100 g l−1 and the enzyme concentration of 0.2 U g−1 of substrate, yielded 37.5 g l−1 of fructose in 20 h at 40°C when A. niger TISTR 3570 inulinase was the biocatalyst. The yield of fructose on inulin was 0.39 g g−1. Under identical conditions, the yeast inulinase afforded 35.3 g l−1 of fructose in 25 h. The fructose yield was 0.35 g g−1 of substrate. The fructose productivities were 1.9 g l−1 h−1 and 1.4 g l−1 h−1 for the mold and yeast enzymes, respectively. After 20 h of reaction, the mold enzyme hydrolysate contained 53% fructose
and more than 41% of initial inulin had been hydrolysed. Using the yeast enzymes, the hydrolysate contained nearly 38% fructose
at 25 h and nearly 36% of initial inulin had been hydrolysed. The A. niger TISTR 3570 inulinases exhibited both endo-inulinase and exo-inulinase activities. In contrast, the yeast inulinases displayed
mainly exo-inulinase activity. The mold and yeast crude inulinases mixed in the activity ratio of 5:1 proved superior to individual
crude inulinases in hydrolysing inulin to fructose. The enzyme mixture provided a better combination of endo- and exo-inulinase
activities than did the crude extracts of either the mold or the yeast individually. 相似文献
5.
Domínguez-Bocanegra AR Ponce-Noyola T Torres-Muñoz JA 《Applied microbiology and biotechnology》2007,75(4):783-791
Phaffia rhodozyma (now Xanthophyllomyces dendrorhous) and Haematococcus pluvialis are known as the major prominent microorganisms able to synthesize astaxanthin natural pigment. Important research efforts
have been made to determine optimal conditions for astaxanthin synthesis. When the focus is on astaxanthin production, the
maximal reported value of 9.2 mg/g cell is obtained within H. pluvialis grown on BAR medium, under continuous illumination (345 μmol photon m−2 s−1) and without aeration. Whereas fermentation by mutated R1 yeast grown on coconut milk produced 1,850 μg/g yeast. However,
when looking at astaxanthin productivity, the picture is slightly different. The figures obtained with P. rhodozyma are rather similar to those of H. pluvialis. Maximal reported values are 170 μg/g yeast per day with a wild yeast strain and 370 μg/g yeast per day with mutated R1 yeast.
In the case of H. pluvialis, maximal values ranged from 290 to 428 μg/g cell per day depending on the media (BG-11 or BAR), light intensity (177 μmol
photon m−2 s−1), aeration, etc. The main aim of this work was to examine how astaxanthin synthesis, by P. rhodozyma and H. pluvialis, could be compared. The study is based on previous works by the authors where pigment productions have been reported. 相似文献
6.
In the wild type strain (stock no. 1227) of Thermoactinomyces vulgaris, as reported earlier [Sinha and Singh (1980) Biochem. J. 190, 457–460], all phosphatase isoenzymes (three alkaline — AlpI,
AlpII and AlpIII, and one acidic — Acp) are present. However, the auxotrophic mutants, the strains 1286 (thi
−), 1279 (nic
−, ura
−) and 1278 (thi
−, ura
−) exhibited two alkaline phosphatase isoenzymes (AlpII and AlpIII), but AlpI was lacking. In the strain 1261 (nic
−, thi
−), only AlpIII was expressed, and AlpI and AlpII isoenzymes were missing. The results suggest that the strains, which require
either thiamine (1286 and 1278) or nicotinamide (1279) for their growth, were AlpI
− mutants; and the strain (1261), which requires both thiamine and nicotinamide for its growth, was AlpI
−/AlpII
− double mutant. There was no direct correlation between uracil auxotrophy and the expression of phosphatases. The uniform
expression of AlpIII and Acp in all the strains, irrespective of their nutrient requirements, suggest that these constitutive
phosphatases are species-specific. The specific activities of the thermophilic acid and alkaline phosphatases were maximum
in the wild type strain (1227) of T. vulgaris. The next in phosphatase activity was the strain 1279 (an AlpI
− mutant), followed by their decrease, in order, in the strains 1286 and 1278 (which were also AlpI
− mutants); while least activity of these enzymes was observed in the obligate thermophile strain 1261 (AlpI
−/AlpII
− double mutant). 相似文献
7.
An endophytic Xylaria sp., having broad antimicrobial activity, was isolated and characterized from Ginkgo biloba L. From the culture extracts of this fungus, a bioactive compound P3 was isolated by bioactivity-guided fractionation and
identified as 7-amino-4-methylcoumarin by nuclear magnetic resonance, infrared, and mass spectrometry spectral data. The compound
showed strong antibacterial and antifungal activities in vitro against Staphylococcus aureus [minimal inhibitory concentrations (MIC) 16 μg·ml−1], Escherichia coli (MIC, 10 μg·ml−1), Salmonella typhia (MIC, 20 μg·ml−1), Salmonella typhimurium (MIC, 15 μg·ml−1), Salmonella enteritidis (MIC, 8.5 μg·ml−1), Aeromonas hydrophila (MIC, 4 μg·ml−1), Yersinia sp. (MIC, 12.5 μg·ml−1), Vibrio anguillarum (MIC, 25 μg·ml−1), Shigella sp. (MIC, 6.3 μg·ml−1), Vibrio parahaemolyticus (MIC, 12.5 μg·ml−1), Candida albicans (MIC, 15 μg·ml−1), Penicillium expansum (MIC, 40 μg·ml−1), and Aspergillus niger (MIC, 25 μg·ml−1). This is the first report of 7-amino-4-methylcoumarin in fungus and of the antimicrobial activity of this metabolite. The
obtained results provide promising baseline information for the potential use of this unusual endophytic fungus and its components
in the control of food spoilage and food-borne diseases. 相似文献
8.
Quaratino D Federici F Petruccioli M Fenice M D'Annibale A 《Antonie van Leeuwenhoek》2007,91(1):57-69
Extracellular laccase from Panus tigrinus CBS 577.79 was produced in a bubble-column reactor using glucose-containing medium supplemented with 2,5-xylidine under conditions of nitrogen sufficiency. The main laccase isoenzyme was purified to apparent homogeneity by ultra-filtration, anion-exchange chromatography and gel filtration that led to a purified enzyme with a specific activity of 317 IU (mg protein)−1 and a final yield of 66%. Laccase was found to be a monomeric protein with a molecular mass of 69.1 kDa, pI of 3.15 and 6.9% N-glycosylation of the high mannose type. Temperature and pH optima were 55°C and 3.75 (2,6-dimethoxyphenol as substrate). At 50 and 60°C, the enzyme half-lives were 281 and 25 min, respectively. The P. tigrinus laccase oxidized a wide range of both naturally occurring and synthetic aromatic compounds: the highest catalytic efficiencies were for 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic) acid and 2,6-dimethoxyphenol (5.99 × 106 and 3.07 × 106 M−1 s−1, respectively). Catalytic rate constants for typical N–OH redox mediators, such as 1-hydroxybenzotriazole (2.6 s−1), violuric acid (8.4 s−1) and 2,2,6,6-tetramethylpiperidin-N-oxide radical (7.8 s−1), were found to be higher than those reported for other high redox potential fungal laccases. 相似文献
9.
The hemolytic activity of the cell-free culture supernatant of Anabaena variabilis OL S1 was investigated using the hemolysis of rabbit erythrocytes as an assay. The culture medium of A. variabilis started to exhibit hemolytic activity at the late exponential growth phase, and maximized at the stationary phase. The hemolytic
toxin is heat-stable and can be extracted in dichloromethane. The hemolytic activities under different temperature, light
intensity and pH showed a high correlation with the cell densities (r=0.965, 0.951, 0.865, respectively), and the optimum condition is 28~30°C, pH 7.5~8.0, light intensity 120 μmol photons m−2s−1. The addition of 10~20 μg mL−1 chloramphenicol, an inhibitor of protein synthesis, exhibited no marked suppression on the hemolytic activity. The supplement
of 1~20 μg mL−1 glycerol increased the hemolytic activity significantly, suggesting that synthesis of hemolysin was dependent on carbohydrate
and lipid metabolism. The spectrum of erythrocyte sensitivity to the hemolysin indicated that rabbit erythrocytes were more
sensitive to the hemolysin than were rat and human erythrocytes. Goldfish and cat erythrocytes were, however, insensitive
to the hemolytic toxin of A. variabilis. 相似文献
10.
He Jun Yu Bing Zhang Keying Ding Xuemei Chen Daiwen 《Indian journal of microbiology》2009,49(2):188-195
The strain of Trichoderma reesei Rut C-30 was subjected to mutation after treatment with N-methyl-N′-nitro-N-nitrosoguanidine (NG) for 6 h followed by UV irradiation for 15 min. Successive mutants showed enhanced cellulase production,
clear hydrolysis zone and rapid growth on Avicel-containing plate. Particularly, the mutant NU-6 showed approximately two-fold
increases in activity of both FPA and CMCase in shake flask culture when grown on basal medium containing peptone (1%) and
wheat bran (1%). The enzyme production was further optimized using eight different media. When a mixture of lactose and yeast
cream was used as cellulase inducer, the mutant NU-6 yielded the highest enzyme and cell production with a FPase activity
of 6.2 U ml−1, a CMCase activity of 54.2 U ml−1, a β-glucosidase activity of 0.39 U ml−1, and a fungal biomass of 12.6 mg ml−1. It deserved noting that the mutant NU-6 also secreted large amounts of xylanases (291.3 U ml−1). These results suggested that NU-6 should be an attractive producer for both cellulose and xylanase production. 相似文献
11.
Qingxin Zhao Sheng Yuan Yuling Zhang Hong Zhu Chuanchao Dai Fang Yang Fengmin Han 《World journal of microbiology & biotechnology》2007,23(8):1057-1064
Pectate lyase A (PelA) of Aspergillus nidulans was successfully expressed in Escherichia coli and effectively purified using a Ni2+-nitrilotriacetate-agarose column. Enzyme activity of the recombinant PelA could reach 360 U ml−1 medium. The expressed PelA exhibited its optimum level of activity over the range of pH 7.5–10 at 50°C. Mn2+, Ca2+, Fe2+, Mg2+ and Fe3+ ions stimulated the pectate lyase activity, but Cu2+ and Zn2+ inhibited it. The recombinant PelA had a V
max of 77 μmol min−1 mg−1 and an apparent K
m of 0.50 mg ml−1 for polygalacturonic acid. Low-esterified pectin was the optimum substrate for the PelA, whereas higher-esterified pectin
was hardly cleaved by it. PelA efficiently macerated mung bean hypocotyls and potato tuber tissues into single cells. 相似文献
12.
T. Zhang 《In vitro cellular & developmental biology. Plant》2007,43(2):91-94
This study investigated the factors affecting in vitro flowering of Perilla frutescens. The shoots regenerated from cotyledonary and hypocotyl explants cultured on Murashige and Skoog (MS) medium supplemented
with benzyladenine (BA) and indole-3-acetic acid, each at 0.5 mg l−1, were excised and transferred to MS medium containing 30 g l−1 of sucrose, 8.25 g l−1 of ammonium nitrate, and 1.0 mg l−1 of BA. After 40 d of culture, 86.2% of shoots flowered and most of which self-fertilized in vitro and produced mature fruits with viable seeds. These seeds were germinated and plants were grown to maturity and flowered
in soil under greenhouse conditions. The in vitro flowering system reported in this study may facilitate rapid breeding of P. frutescens and offers a model system for studying the physiological mechanism of flowering. 相似文献
13.
14.
Two oligosaccharides, a heptasaccharide (HS) and an octasaccharide (OS), isolated from Paris polyphylla var. yunnanensis, stimulated the growth and saponin accumulation of Panax ginseng hairy roots at 5–30 mg l−1. HS and OS at 30 mg l−1, fed separately to hairy root cultures at 10 days post-inoculation, increased the root biomass dry weight by more than 70%
to ∼20 g l−1 from 13 g l−1 and the total saponin content of roots by more than 1-fold to ∼3.5% from 1.6% (w/w). The results suggest that the two oligosaccharides
may have plant growth-regulatory activity in plant tissue cultures. 相似文献
15.
Light irradiation had remarkable effects on callus growth of Oldenlandia affinis with an optimum intensity of 35 μmol m−2 s−1. Biosynthesis of kalata B1, the main cyclic peptide in O. affinis, was induced and triggered with rising irradiation intensities. The highest concentration of kalata B1, 0.49 mg g−1 DW characterised by the maximum productivity of 3.88 μg per litre and day was analysed at 120 μmol m−2 s−1, although callus growth was repressed. The light saturation point was established to be 35 μmol m−2 s−1, where kalata B1 productivity was in a similar order (3.41 μg per day) due to the higher growth index. O. affinis suspension cultures were shown to accumulate comparable specific kalata B1 concentrations in a delayed growth associated
production pattern. These were dependent on irradiation intensity (0.16 mg g−1 at 2 μmol m−2 s−1; 0.28 mg g−1 at 35 μmol m−2 s−1). The batch cultivation process resulted in a maximum productivity of 27.30 μg per litre and day with culture doubling times
of 1.16 d−1. Submers operation represented a 8-fold product enhancement compared to callus cultivation. 相似文献
16.
Jaya Ram Simkhada Hyo Jeong Lee So Young Jang Seung Sik Cho Eun Jung Park Jae Kyung Sohng Jin Cheol Yoo 《Biotechnology letters》2009,31(3):429-435
A 60 kDa phospholipase D (PLD) was obtained from Streptomyces olivochromogenes by one-step chromatography on Sepharose CL-6B. Maximal activity was at pH 8 and 75°C and the enzyme was stable from pH 7
to 13 and from 55 to 75°C. Thermal and pH stability with temperature optimum of the enzyme were highest among Streptomyces PLDs reported so far. The activity was Ca2+-dependent and enhanced by detergents. The Km and Vmax values for phosphatidylcholine were 0.6 mM and 650 μmol min−1 mg−1, respectively. In addition, the enzyme also revealed transphosphatidylation activity, which was optimum at pH 8 and 50°C.
The first 15 amino acid residues of the N terminal sequence were ADYTPGAPGIGDPYY, which are significantly different from the
other known PLDs. The enzyme may therefore be a novel PLD with potential application in the lipid industry. 相似文献
17.
Fábio C. Sampaio Janaína T. de Faria Flávia M. Lopes Passos Attilio Converti Luis Antônio Minin 《Journal of industrial microbiology & biotechnology》2009,36(2):293-300
Xylose reductase (XR) is the enzyme that catalyzes the first step of xylose metabolism. Although XRs from various yeasts have
been characterized, little is known about this enzyme in Debaryomyces hansenii. In the present study, response surface analysis was used to determine the optimal conditions for D. hansenii UFV-170 XR activity. The influence of pH and temperature, ranging from 4.0 to 8.0 and from 25 to 55°C, respectively, was
evaluated by a 22 central composite design face-centered. The F-test (ANOVA) and the Student’s t test were performed to evaluate the statistical significance of the model and the regression coefficients, respectively.
The NADPH-dependent XR activity varied from 0.502 to 2.53 U mL−1, corresponding to 0.07–0.352 U mg−1, whereas the NADH-dependent one was almost negligible. The model predicted with satisfactory correlation (R
2 = 0.940) maximum volumetric activity of 2.27 U mL−1 and specific activity of 0.300 U mg−1 at pH 5.3 and 39°C, which were fairly confirmed by additional tests performed under these conditions. The enzyme proved very
stable at low temperature (4°C), keeping its activity almost entirely after 360 min, which corresponded to the half-time at
39°C. On the other hand, at temperatures ≥50°C it was lost almost completely after only 20 min. 相似文献
18.
Meiru Li Hongqing Li Huawu Jiang Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2008,92(2):173-181
Jatropha curcas contains high amounts of oil in its seed and has been considered for bio-diesel production. A transformation procedure for
J. curcas has been established for the first time via
Agrobacterium tumefaciens infection of cotyledon disc explants. The results indicated that the efficiency of transformation using the strain LBA4404
and phosphinothricin for selection was an improvement over that with the strain EHA105 and hygromycin. About 55% of the cotyledon
explants produced phosphinothricin-resistant calluses on Murashige and Skoog (MS) medium supplemented with 1.5 mg l−1 benzyladenine (BA), 0.05 mg l−1 3–indolebutyric acid (IBA), 1 mg l−1 phosphinothricin and 500 mg l−1 cefotaxime after 4 weeks. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing
1.5 mg l−1 BA, 0.05 mg l−1 IBA, 0.5 mg l−1 gibberellic acid (GA3), 1 mg l−1 phosphinothricin and 250 mg l−1 cefotaxime, and about 33% of the resistant calli differentiated into shoots. Finally, the resistant shoots were rooted on
1/2 MS media supplemented with 0.3 mg l−1 IBA at a rate of 78%. The transgenic nature of the transformants was demonstrated by the detection of β-glucuronidase activity
in the primary transformants and by PCR and Southern hybridization analysis. 13% of the total inoculated explants produced
transgenic plants after approximately 4 months. The procedure described will be useful for both, the introduction of desired
genes into J. curcas and the molecular analysis of gene function. 相似文献
19.
A novel phytase gene appA, with upstream and downstream sequences from Citrobacter amalonaticus CGMCC 1696, was cloned by degenerate polymerase chain reaction (PCR), and thermal asymmetric interlaced (TAIL) PCR and was
overexpressed in Pichia pastoris. Sequence analysis revealed one open reading frame that consisted of 1311 bp encoding a 436–amino-acid protein, which had
a deduced molecular mass of 46.3 kDa. The phytase appA belongs to the histidine acid phosphatase family and exhibits the highest
identity (70.1%) with C. braakii phytase. The gene was overexpressed in P. pastoris. The secretion yield of recombinant appA protein was accumulated to approximately 4.2 mg·mL−1, and the enzyme activity level reached 15,000 U·mL−1, which is higher than any previous reports. r-appA was glycosylated, as shown by Endo H treatment. r-appA was purified and
characterized. The specific activity of r-appA for sodium phytate was 3548 U·mg−1. The optimum pH and temperature for enzyme activity were 4.5 and 55°C, respectively. r-appA was highly resistant to pepsin
or trypsin treatment. This enzyme could be an economic and efficient alternative to the phytases currently used in the feed
industry. 相似文献
20.
Meiru Li Hongqing Li Huawu Jiang Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2008,93(3):249-255
Broussonetia papyrifera is well-known for its bark fibers, which are used for making paper, cloth, rope etc. This is the first report of a successful
genetic transformation protocol for B. papyrifera using Agrobacterium tumefaciens. Callus was initiated at a frequency of about 100% for both leaf and petiole explants. Shoots formed on these calli with
a success rate of almost 100%, with 14.08 and 8.36 shoots regenerating from leave-derived and petiole-derived callus, respectively.
For genetic transformation, leaf explants of B. papyrifera were incubated with A. tumefaciens strain LBA4404 harboring the binary vector pCAMBIA 1301 which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene (gus-int) as a reporter gene. Following co-cultivation, leaf explants were cultured on Murashige and Skoog (Physiol Plant 15:473,
1962) (MS) medium supplemented with 1.5 mg l−1 benzyladenine (BA) and 0.05 mg l−1 indole-3-butyric acid (IBA) (CI medium) containing 5 mg l−1 hygromycin and 500 mg l−1 cefotaxime, in the dark. Hygromycin-resistant calli were induced from leaf explants 3 weeks thereafter. Regenerating shoots
were obtained after transfer of the calli onto MS medium supplemented with 1.5 mg l−1 BA, 0.05 mg l−1 IBA, and 0.5 mg l−1 gibberellic acid (GA3) (SI medium), 5 mg l−1 hygromycin and 250 mg l−1 cefotaxime under fluorescent light. Finally, shoots were rooted on half strength MS medium (1/2 MS) supplemented with 10 mg l−1 hygromycin. Transgene incorporation and expression was confirmed by PCR, Southern hybridisation and histochemical GUS assay.
Using this protocol, transgenic B. papyrifera plants containing desirable new genes can be obtained in approximately 3 months with a transformation frequency as high as
44%. 相似文献