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PubMed2000

Purification and characterization of a mannan-binding lectin specifically expressed in corms of saffron plant (Crocus sativus L.).

Despite the economical interest of Crocus sativus, its biochemistry has been poorly studied. Herein, we have isolated a lectin present in saffron corm by gel-filtration, anion-exchange, and reversed-phase chromatography. One- and two-dimensional PAGE, MALDI-MS, and N-terminal amino acid sequence analyses indicated that the native protein forms noncovalently linked aggregates of about 80 kDa apparent molecular mass, mainly composed of two charged heterogeneous (pI's, 6.69-6.93) basic subunits of approximately 12 kDa. Their N-terminal sequences shared 25% similarity and were homologous to the N- and C-terminal domains of monocotyledonous mannose-binding lectins, respectively. An additional polypeptide of around 28 kDa apparent molecular mass was also detected, probably corresponding to a precursor processed into two mature subunits. In addition, the N-terminal domain subunit exhibited 56% similarity with curculin, a sweet protein with taste-modifying activity. The native lectin specifically interacts with a yeast mannan and is a major corm protein specifically expressed in this organ.

Amino Acid Sequence↗
PubMed2000

The cytolytic effect of a glycoconjugate extracted from corms of saffron plant (Crocus sativus) on human cell lines in culture.

Corms of Crocus sativus L. (Iridaceae) contain a glycoconjugate that shows cytotoxic activity on tumoral cells in culture. Studies of intracellular calcium fluctuations, and release of lactate dehydrogenase in human cervical epitheloid carcinoma cells, showed that this compound caused plasma membrane damage, allowing movements of both calcium and macromolecules, and leading to cell lysis. Analysis of DNA fragmentation showed that cell death was not mediated by apoptosis. This molecule is active against human tumoral cells derived from fibrosarcoma, cervical epithelioid carcinoma and breast carcinoma, with IC50 values of 7, 9 and 22 micrograms/ml, respectively. The proteoglycan is about 8 times more cytotoxic for malignant cells than for their normal counterparts. In addition, 100 micrograms/ml of proteoglycan produced 50% in vitro lysis of normal human erythrocytes, whereas 320 micrograms/ml induced about 60% cell death on cultured human hair follicles. Altogether, these results suggests a distinctive cytotoxic activity of this molecule on different human cell types.

Antineoplastic Agents, Phytogenic↗
PubMed2001

Authentic identification of stigma Croci (stigma of Crocus sativus) from its adulterants by molecular genetic analysis.

Stigma Croci, stigma of Crocus sativus L., is a precious traditional Chinese medicine, which is commonly used to activate blood circulation and to dissipate blood stasis. Three plant species, Carthamus tinctorius L., Hemerocallis fulva (L.) L. and Hemerocallis citrina Baroni, could carry the name Stigma Croci in the commercial markets of South East Asia. However, C. sativus is the only one that has proven its effectiveness, while the others could act as adulterants. The authentic identification of C. sativus on the market is difficult. By using molecular genetic method, the spacer domains of 5S-rRNA were cloned from the genomic DNAs that were isolated from C. sativus, C. tinctorius, H. fulva and H. citrina. The cDNAs encoding the spacer domains, about 300 to 500 bp, were sequenced. The nucleotide sequences of these four species showed great diversity, which could serve as markers for authentic identification of Stigma Croci to distinguish from its substitution and counterfeit.

Drugs, Chinese Herbal↗
PubMed1997

In vitro formation of crocetin glucosyl esters by Crocus sativus callus extract.

Callus extracts of Crocus sativus exhibited the ability to transform all-trans-crocetin into its related glycosides between pH 7.0 and 7.6 in the presence of uridine-diphospho-glucose. The reactions involved the step-wise addition of a glucose moiety to a free carboxyl function and the 1-->6 addition of a glucose moiety to a glucosyl ester function. The kinetics of synthesis for each glycoside seemed to indicate that two distinct glucosyl transferases were implicated in the synthesis of crocin, all-trans-crocetin di-(beta-D-gentiobiosyl) ester.

Anticarcinogenic Agents↗
PubMed2000

Crocus sativus lectin recognizes Man3GlcNAc in the N-glycan core structure.

Crocus sativus lectin (CSL) is one of the truly mannose-specific plant lectins that has a unique binding specificity that sets it apart from others. We studied sugar-binding specificity of CSL in detail by a solution phase method (fluorescence polarization) and three solid phase methods (flow injection, surface plasmon resonance, and microtiter plate), using a number of different glycopeptides and oligosaccharides. CSL binds the branched mannotriose structure in the N-glycan core. Substitution of the terminal Man in the Manalpha(1-3)Man branch with GlcNAc drastically decreases binding affinity much more than masking of the terminal Man in the Manalpha(1-6)Man branch. Most interestingly, the beta-Man-linked GlcNAc in N-glycan core structure contributes greatly to the binding. The effect of this GlcNAc is so strong that it can substantially offset the negative effect of substitution on the nonreducing terminal Man residues. On the other hand, the GlcNAc that is usually attached to Asn in N-glycans and the l-Fuc linked at the 6-position of the GlcNAc are irrelevant to the binding. A bisecting GlcNAc neither contributes to nor interferes with the binding. This unique binding specificity of CSL offers many possibilities of its use in analytical and preparative applications.

Animals↗
PubMed2004

Accidental poisoning with autumn crocus.

We describe a case of a 43-yr-old female with severe multiorgan injury after accidental poisoning with Colchicum autumnale, which was mistaken for wild garlic (Allium ursinum). Both plants grow on damp meadows and can be confused in the spring when both plants have leaves but no blossoms. The autumn crocus contains colchicine, which inhibits cellular division. Treatment consisted of supportive care, antibiotic therapy, and granulocyte-directed growth factor. The patient was discharged from the hospital after three weeks. Three years after recovery from the acute poisoning, the patient continued to complain of muscle weakness and intermittent episodes of hair loss.

Accidents↗
PubMed1999

Effects of long-term treatment of colon adenocarcinoma with crocin, a carotenoid from saffron (Crocus sativus L.): an experimental study in the rat.

We used an experimental model in the rat to examine the effects of long-term treatment with crocin, a glycosylated carotenoid from the stigmas of the saffron crocus, on colon cancer. BD-IX rats were divided into four groups: Groups G1 and G2, designated "cancer groups," were used to study the effects of crocin on the progression of colon cancer, and Groups G3 and G4, designated "toxicity groups," were used to study the effects of the treatment on metabolic processes and the parenchyma. DHD/K12-PROb cells were injected subcutaneously into the chest of Group G1 and G2 animals. From 1 to 13 weeks after inoculation, animals in Groups G2 and G4 received a weekly injection of crocin (400 mg/kg body wt s.c.). Animals in Groups G1 and G3 received no treatment. In addition, lines of animal and human colon adenocarcinoma cells (DHD/K12-PROb and HT-29) were used to perform assays in vitro to examine the cytotoxicity of crocin. Life span was extended and tumor growth was slower in crocin-treated female rats, but no significant antitumor effect was found in male rats. Acute tubular necrosis was found in all kidney samples from crocin-treated animals, but slight signs of nephrotoxicity were found by biochemical analysis of the serum. In assays in vitro, crocin had a potent cytotoxic effect on human and animal adenocarcinoma cells (HT-29 and DHD/K12-PROb cells, 50% lethal dose = 0.4 and 1.0 mM, respectively). Treated cells exhibited a remarkable loss of cytoplasm and wide cytoplasmic vacuole-like areas. In conclusion, long-term treatment with crocin enhances survival selectively in female rats with colon cancer without major toxic effects. The effects of crocin might be related to its strong cytotoxic effect on cultured tumor cells.

Adenocarcinoma↗
PubMed1994

Effects of Crocus sativus L. on the ethanol-induced impairment of passive avoidance performances in mice.

The acute effects of an alcohol extract of Crocus sativus L. (CS-extract) were studied on learning and memory in step through (ST) and step down (SD) tests in normal as well as in learning- and memory-impaired mice. A single oral administration of CS-extract had no effects on memory registration, consolidation or retrieval in normal mice. CS-Extract reduced the ethanol-induced impairment of memory registration both in ST and SD tests and the ethanol-induced impairment of memory retrieval in SD test. CS-Extract decreased the motor activity (MA) and prolonged the sleeping time induced by hexobarbital. These results suggested that CS-extract ameliorates the impairment effects of ethanol on learning and memory processes, and possesses a sedative effect.

Administration, Oral↗
PubMed2000

A glycoconjugate from corms of saffron plant (Crocus sativus L.) inhibits root growth and affects in vitro cell viability.

A glycoconjugate has been characterized from saffron corms (Crocus sativus L.) that inhibits the growth of roots of Nicotiana tabacum and Arabidopsis thaliana, at concentrations ranging from 1-100 micrograms m-3. Roots of seedlings grown in the presence 0.1 microgram m-3 glycoconjugate showed bulging of epidermal cells, whereas at 10 micrograms m-3, roots were completely devoid of hairs. At 100 micrograms m-3 glycoconjugate the cell walls of the root vascular tissues were thicker and, overall, the vascular tissue was enlarged. In addition, this glycan is cytotoxic to isolated tobacco cells and protoplasts, with 50% cell death induced by 0.5 and 2 micrograms m-3 glycoconjugate, respectively. Morphological and biochemical changes induced by the exposure to the glycoconjugate included cell size decrease, loss of regular cell shape, cytoplasm collapse, and release of intracellular proteins. This molecule at low concentrations (0.1 microgram m-3) mimics the effects of Yariv phenylglycosides and of mutant Arabidopsis which present defective or missing arabinogalactan-proteins (AGPs) in roots, indicating the glycoconjugate might interact with cell surface AGPs.

Arabidopsis↗
PubMed1990

Characterization of the platelet aggregation inducer and inhibitor isolated from Crocus sativus.

Bulbs of Crocus sativus variety Cartwrightianus were found to contain both a platelet aggregation inducer and inhibitor. The aggregating factor has a Mr of 42 kDa estimated by a Sephadex G75 column and SDS-polyacrylamide slab gel electrophoresis. It was found to lack enzymatic activity such as proteinase, esterase and acid or alkaline phosphatase. The inhibitory factor was also purified to homogeneity by different chromatographic techniques and shows a Mr of 27 kDa as it was estimated by Biogel P30 column and SDS-polyacrylamide slab gel electrophoresis. It was found to possess strong proteinase activity.

Acid Phosphatase↗
PubMed2005

A modified, economic, sensitive method for measuring total antioxidant capacities of human plasma and natural compounds using Indian saffron (Crocus sativus).

BACKGROUND: Free radicals are involved in various human diseases that can possibly be prevented by antioxidants. There are many but rather expensive methods to determine total antioxidant capacity of human plasma (for endogenous antioxidant levels) or plant extracts/natural compounds (for antioxidant potential in terms of radical inhibiting or scavenging properties). We describe a simple, fast and economical 'crocin assay' using the Indian spice saffron. METHODS: In crocin assay, the extent of bleaching of crocin, a carotenoid from saffron, by peroxyl radicals generated by thermal decomposition of azo-initiator was measured. We examined its applicability to clinical samples and plant extracts. RESULTS: The cost of Indian saffron is almost 38 times less per unit dry weight compared to the 'Sigma' saffron. Yet, it gives 26 times better yield of crocin than that from 'Sigma' saffron. It was also shown that Indian saffron is rich in crocin. The total antioxidant capacity (TAC) values of human plasma from normal, healthy individuals, using Sigma as well as Indian crocin, expressed in terms of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) equivalent antioxidant capacity (TEAC), were comparable. We have also demonstrated that crocin assay can be used for clinical samples such as plasmas from healthy and diabetic individuals. The antioxidant potentials, TEAC, of plant extracts and pure natural compounds by Indian and Sigma crocin assays were similar. Addition of uric acid to plasma induced a concentration-dependent response. The assay was compared to standard radical scavenging 1,1'-diphenyl-2-picrylhydrazyl (DPPH) assay and was found to match well, showing better sensitivity and hence validates this assay for natural compounds and clinical samples. CONCLUSIONS: Development of crocin assay using the Indian saffron is economical and sensitive method for measurement of total antioxidant capacities from human plasma as well as natural compounds and plant extracts.

Adult↗
PubMed2002

Catalytic properties of three L-lactate dehydrogenases from saffron corms (Crocus sativus L).

Three L-lactate dehydrogenase isoenzymes were detected in saffron corms, using potassium ferricyanide as the electron acceptor. Their pH optima were 5.5, 7.5 and 9.5, respectively. All three dehydrogenases were substrate-inhibited by ferricyanide, but at different concentrations; maximum enzymatic activity was observed for 250, 100 and 600 microM ferricyanide, at pH 5.5, 7.5 and 9.5, respectively. Catalytic efficiency, calculated per mg corm extract protein, was 1.9, 1.0 and 0.4 min(-1), respectively at pH 5.5, 7.5 and 9.5. Pseudo first order rate constant was also different under the three pH conditions. Malate was an inhibitor for the isoenzyme active at pH 9.5, but had no effect on the others.

Catalysis↗
PubMed2003

Production of crocin using Crocus sativus callus by two-stage culture system.

Saffron callus was grown in a two-stage culture on B5 medium supplemented with casein hydrolysate (300 mg l(-1)) at 22 degrees C in dark with naphthalene acetic acid (2 mg l(-1)) and 6-benzyladenine (1 mg l(-1)) to give maximum biomass (16 g dry wt l(-1)), and with indole 3-acetic acid (2 mg l(-1)) and 6-benzyladenine (0.5 mg l(-1)) for crocin formation. The maximum crocin production (0.43 g l(-1)) was achieved by this two-stage culture method, which was three times that by a one-stage method.

Bioreactors↗