Purification and characterization of a lectin from endophytic fungus Fusarium solani having complex sugar specificity

Khan, Feroz; Ahmad, Absar; Khan, M. Islam

doi:10.1016/j.abb.2006.10.019

Cited by 59 publications

(48 citation statements)

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“…A 26 kDa dimeric lectin from endophytic fungus Fusarium solani was purified and characterized earlier, and showed specificity for N-linked as well as O-linked glycotopes (10). This communication elaborates studies on the conformation stability of the lectin from F. solani by chemical, thermal and pH dependent denaturation.…”

Section: Introductionmentioning

confidence: 78%

“…Cultivation of F. solani (NCIM 1330) and purification of lectin was carried out as described earlier (10). Protein concentrations were determined according to Bradford et al (11) using BSA as a standard.…”

Section: Purification Of F Solani Lectinmentioning

confidence: 99%

“…3E). In the hemagglutination assays, the F. solani lectin was found to have hemagglutinating activity in a broad range of pH varying from 2 -12 (10).…”

Section: Ph-dependent Denaturationmentioning

confidence: 99%

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Chemical, thermal and pH‐induced equilibrium unfolding studies of Fusarium solani lectin

2007

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SummaryThe effect of urea, guanidine thiocyanate, temperature and pH was studied on the conformational stability of Fusarium solani lectin. Equilibrium unfolding with chemical denaturants showed that the lectin was least stable at pH 12 and maximally stable at pH 8.0 near its pI (8.7). Guanidine thiocyanate (the concentration of denaturant at which the protein is half folded, D 1/2 ¼ 0.49 M at pH 12) was found to be an eight times stronger denaturant than urea (D 1/2 ¼ 3.88 M at pH 12). The unfolding curves obtained with fluorescence and CD measurements showed good agreement indicating a monophasic nature of unfolding and excluded the possibility of formation of any stable intermediate. The effect of pH on the lectin was found to be unusual as at acidic pH, the lectin showed a flexible tertiary structure with pronounced secondary structure, and retained its hemagglutinating activity. On the other hand, the lectin did not show any loss of conformation or activity upto 708C for 15 min. Moreover, thermal denaturation did not result in the aggregation or precipitation of the protein even at high temperatures. Thermal denaturation was also carried out in the presence of a low concentration of guanidine thiocyanate. Change in the enthalpy of transition (DH m ) varied linearly with transition temperature (T m ), which indicated that the heat capacity (DC p ¼ 3.95 kJ Á mol 71 Á K 71) of the lectin remained constant during the unfolding.

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Section: Introductionmentioning

confidence: 78%

Section: Purification Of F Solani Lectinmentioning

confidence: 99%

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Chemical, thermal and pH‐induced equilibrium unfolding studies of Fusarium solani lectin

2007

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“…Moreover, the lectin from Fusarium solani had blood specificity with neuraminidase or pronase treated human ABO type blood and no blood specificity with trypsin treated or untreated human blood [17]. From these results, it was suggested that lectin had different specificity to other enzymes according to red blood cell features, type of enzyme and its treatment.…”

Section: Discussionmentioning

confidence: 96%

Comparison of Biochemical Characterization of Korean and Chinese Mung Bean Lectin

Roh

2014

Journal of Life Science

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The lectins were separated from Korean and Chinese mung bean seeds finally via chromatography using Sephadex G-100 and their biochemical features were studied and compared. They showed no hemagglutination with human red blood cells regardless of trypsin treatment and showed hemagglutination with only trypsin treated rabbit red blood cells. The molecular weights of two lectins were identified as 54 kDa and 28 kDa by SDS-PAGE. It was found that while the optimal reaction temperature of the lectin from Korean mung bean was 60℃, that of the lectin from Chinese mung bean seeds was 50℃. It was found also that the most thermal stable temperature of the seed lectin from Korean mung bean seeds was 50℃ and the lectin from Chinese mung bean was 40-50℃. The lectin from Korean mung bean seeds showed the highest activity at pH 3.2 and the lectin from Chinese mung bean showed the highest activity at pH 6.2. It was identified that when treating a denaturant, thiourea and guanidine-HCl resulted in no hemagglutination, so they induced denaturalization. It was identified also that there was no hemagglutination with urea, so it did not induced denaturalization. They showed no septicity to 6 types of carbohydrates including D-glucose. In addition, the lectins from the two mung bean seed had specificity to metal ions.Key words : Biochemical characterization, Korean and Chinese mung bean seed, lectin *Corresponding author *Tel : +82-53-580-5207, Fax : +82-53-580-5164 *E-mail : rks@kmu.ac.kr This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. IntroductionMung bean (Phaseolus radiatus L. var.) is an annual plant which belongs to Leguminosae and is originated from India and cultivated also in Korea. Length of its stem is 60~80 cm and brown pili are spread on the whole of plant. Its pod is 5~6 cm in length, has slender and long shape with rough pili and projections, and contains 10-15 fruits. As it is highly nutritious, containing about 60% of starch and 21% of protein as main ingredients, it serves an important nutrition source to animals as well as humans and is also used as a major protein source due to its higher fiber content and lower fat content [28].Lectin is one of natural proteins cohering reversibly to specific monosaccharide and polysaccharide and has specific binding affinity to carbohydrates [30] It is widely distributed across plants, animals and microorganisms in nature [26]. Plant lectin is mainly isolated from dried seeds, but exists also in leaves, stems, roots, and tubers [39]. Particularly it is rich in seeds of leguminous plants among them [5,27].The lectin is synthesized in ribosome of plant cells, transferred to secretion system in form of glycoprotein, accumulated highly in vacuoles and cell walls [6], and then used in growth of young plants. In additions it serves...

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“…They also have been widely used to distinguish between cell types [5,10]. Lectins are also found in microorganisms, plants and animals and have attracted great interest due to their varied physiological roles in cell agglutination [13], anti-tumour [15,16], immunomodulatory [22], antifungal [12] and antiviral effects [29]. Furthermore, lectin-mediated drugs have been acquired to target specific cells and some lectins with anti-proliferative properties were isolated and characterized from different parts of the plant, like; seeds [14], leaves [19] and roots [27,31].…”

Section: Introductionmentioning

confidence: 99%

Isolation, characterization and production of a new recombinant lectin protein from leguminous plants

Patel¹

2014

Bio Chem Comp

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Lectins are present in microorganisms, plants and animals and have attracted great interest due to their varied physiological roles in cell agglutination, anti-tumour, immunomodulatory, antifungal and antiviral effects. Legume lectins are important to the pharmaceuticals but they are produced in low amounts in the plant seeds. Moreover, the genes controlling these proteins are conditionally active, i.e., they work under specific circumstances and not in regular manner. Looking into these limitations, we aimed this work to produce a recombinant lectin for pharmaceutical use especially for the treatment of cancer. Three different legume plant species were collected from Aseer region of Kingdom of Saudi Arabia viz., Acacia seyal, Pisum sativum (wild type) and Pisum sativum (Pea). The plant tissues were subjected to RNA extraction, the extracted RNA was used for lectin gene amplification using specific primers. Cloning, subcloning of the Acacia 400bp gene was carried out and in vitro transcription, combined with protein purification was undertaken. The cytotoxicity of the recombinant lectin was performed on two cell lines such as breast cancer MCF-7 and liver cancer HepG-2 cells. Our study resulted in the observation of two amplicones with all the three examined species, the amplicone molecular sizes were 800 and 400bp. The 400bp amplicone was excised from the agarose gel, purified and sequenced. The sequence analysis revealed that the nucleotide sequence belongs to lectin gene. The sequence analysis revealed that the lectin gene isolated from Pisum sativum (wild plant) is similar to the Pisum sativum lect1 with identity 90%, whereas, lectin isolated from Pisum (pea) showed similarity of 91% with the other lectins. On the other hand, Acacia lectin showed similarity with Lotus japonicus nod factor binding lectin gene with identity of 95%. Thus we conclude that new lectin protein of 17 and 15 kDa was produced that can be used by pharmaceutical industries.

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Purification and characterization of a lectin from endophytic fungus Fusarium solani having complex sugar specificity

Cited by 59 publications

References 34 publications

Chemical, thermal and pH‐induced equilibrium unfolding studies of Fusarium solani lectin

Chemical, thermal and pH‐induced equilibrium unfolding studies of Fusarium solani lectin

Comparison of Biochemical Characterization of Korean and Chinese Mung Bean Lectin

Isolation, characterization and production of a new recombinant lectin protein from leguminous plants

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