Purification and Characterization of a Highly Thermostable Chitinase from the Stomach of the Red ScorpionfishScorpaena scrofawith Bioinsecticidal Activity toward Cowpea WeevilCallosobruchus maculatus(Coleoptera: Bruchidae)

Laribi-Habchi, Hassiba; Dziril, Maya; Badis, Abdelmalek; Mouhoub, Samia; Mameri, N.

doi:10.1271/bbb.120344

Cited by 20 publications

(4 citation statements)

References 45 publications

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“…EDC did not inhibit the activity of the enzyme, suggesting that the glutamic acid residue in the active site was not accessible to EDC. This behavior was similar to that observed for the purified chitinase ScChi50 from the stomach of red scorpionfish Scorpaena scrofa already described by the authors [24]. Chitinase activity was, however, strongly inhibited by Ag 2+ and by Cd 2+ , completely inhibited by Hg 2+ and by Hg + , and moderately inhibited by Fe 2+ .…”

Section: Chemical Modification and Effect Of Metallic Ions On The Actsupporting

confidence: 89%

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria

Laribi-Habchi

Bouanane‐Darenfed

Drouiche

et al. 2015

International Journal of Biological Macromolecules

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Section: Chemical Modification and Effect Of Metallic Ions On The Actsupporting

confidence: 89%

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria

Laribi-Habchi

Bouanane‐Darenfed

Drouiche

et al. 2015

International Journal of Biological Macromolecules

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“…Chitinase (EC 3.2.1.14) was purified from the stomach of red scorpion fish by ammonium sulfate precipitation followed by chromatography gel filtration Sephadex G‐75 (Pharmacia, Munich, Germany) and exchange ionic chromatography Mono Q‐Sepharose (Pharmacia, Freiberg, Germany) (Laribi‐Habchi et al . ).…”

Section: Methodsmentioning

confidence: 97%

Efficacy of Crude and Purified Chitinases (SsChi50) Extracted from Offal Red Scorpion Fish in Biological Control of Chickpea Weevil (C allosobruchus maculatus L.)

Laribi-Habchi

Biche

Drouiche

et al. 2014

Journal of Food Processing and Preservation

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In the tropical zone, Callosobruchus maculatus (Coléoptere: Bruchidae) is one of the most important pests of seeds of chickpea (Cicer arietinum). The infestation starts in the fields and continues in the stocks where the damage can be considerable in the absence of any protective measures. The importance of this damage justifies the development of appropriate control methods. In the context of biological control as an alternative to chemical control, the crude and purified chitinases SsChi50 extracted from the red scorpion fish offal were tested for their possible role as insecticide on the stored-product insect Ca. maculatus at various doses, respectively (3, 6, 9, 12, 15, 18, 21 and 24%), (1, 1.5, 2, 2.5 and 3%). The biotests were conducted in laboratory conditions (temperature and relative humidity) maintained at 28C and 75%, respectively. The results showed that the crude and purified chitinases (SsChi50) tested possess insecticidal activity against Ca. maculatus. The crude and purified chitinases at doses of 24 and 3%, respectively, exhibited great biocidal power against insects as we experienced 100% mortality after only 1 h of exposure to treatment. The 50% lethal dose (LD) at each time of exposure was estimated. This allows us to move forward so that the crude and purified chitinases extracted from the red scorpion fish offal tested can be used as bioinsecticide for the control of Ca. maculatus populations of pests of stored foodstuffs. PRACTICAL APPLICATIONSWe develop a novel bioinsecticide which consists of crude and purified chitinases extracted from fish offal (scorpion fish). The biotests conducted in laboratory conditions revealed that this crude and purified chitinases extract has 100% insecticide activity after 1 h contact at doses of 24 and 3%, respectively. The originality of this work therefore lies in the fact that there are no studies on the use of chitinases extracted from marine biomass as bioinsecticide.

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“…The protein bands were visualized with Coomassie Brilliant Blue R-250 (Bio-Rad Laboratories, Inc., Hercules, CA, USA) staining. Zymography analysis was monitored as reported by Laribi-Habchi et al [14,17] using chitin azure as substrate with slight modification. Discontinuous substrate SDS-PAGE (Zymogram analysis) was performed with a 4% stacking gel, except that 1 mg/ml of heat-denatured chitin azure was incorporated into the 12% separation gel.…”

Section: Protein Quantification Electrophoresis and Mass Spectrometrymentioning

confidence: 99%

Production of ChiA-Pt70, a new organic solvent-tolerant extracellular chitinase from Paenibacillus timonensis strain LK-DZ15

Yahiaoui

Bouacem

HARIR

et al. 2021

Proceedings of MOL2NET'21, Conference on Molecular, Biomedical &Amp; Computational Sciences and Engineering, 7th Ed.

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A new extracellular chitinase (ChiA-Pt70) was produced and purified from a newly isolated Paenibacillus timonensis strain LK-DZ15. The maximum chitinase activity recorded after 44-h of incubation at 30°C was 11,500 U/mL. Pure enzyme was obtained after ammonium sulphate precipitation (40-70%) followed by sequential column chromatographies on fast performance liquid chromatography (FPLC) and high performance liquid chromatography (HPLC). Based on matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis, the purified enzyme is a monomer with a molecular mass of 70,166.11 kDa. The sequence of the 25 NH2terminal residues of the mature ChiA-70 showed high homology with Paenibacillus GH-18 chitinases family. Optimal activity was achieved at pH 4.5 and 80°C. The pure enzyme was completely inhibited by p-chloromercuribenzoic acid (p-CMB) and N-ethylmaleimide (NEM). Chitinase activity was high on colloidal chitin, chitin azure, glycol chitin, glycol chitosane, chitotriose, and chito-oligosaccharide while it did not hydrolyse chitibiose and amylose. Furthermore, thin-layer chromatography (TLC) analysis from enzymatic catalyzed hydrolysis of chitin-oligosaccharides showed that ChiA-Pt70 acted as an endo-splitting enzyme. Its Km and kcat values were 0.611 mg colloidal chitin/mL and 87,800 s -1 , respectively. Interestingly, its catalytic efficiency was higher than those of chitinases ChiA-Mt45 from Melghiribacillus thermohalophilus strain Nari2A T , ChiA-Hh59 from Hydrogenophilus hirchii strain KB-DZ44, Chitodextrinase ® from Streptomyces griseus, and N-acetyl-β-glucosaminidase ® from Trichoderma viride. Therefore, ChiA-Pt70 exhibited remarkable biochemical properties suggesting that it is suitable for the enzymatic degradation of chitin.

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Purification and Characterization of a Highly Thermostable Chitinase from the Stomach of the Red ScorpionfishScorpaena scrofawith Bioinsecticidal Activity toward Cowpea WeevilCallosobruchus maculatus(Coleoptera: Bruchidae)

Cited by 20 publications

References 45 publications

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria

Efficacy of Crude and Purified Chitinases (SsChi50) Extracted from Offal Red Scorpion Fish in Biological Control of Chickpea Weevil (C allosobruchus maculatus L.)

Production of ChiA-Pt70, a new organic solvent-tolerant extracellular chitinase from <em>Paenibacillus timonensis</em> strain LK-DZ15

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Purification and Characterization of a Highly Thermostable Chitinase from the Stomach of the Red ScorpionfishScorpaena scrofawith Bioinsecticidal Activity toward Cowpea WeevilCallosobruchus maculatus(Coleoptera: Bruchidae)

Cited by 20 publications

References 45 publications

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria

Efficacy of Crude and Purified Chitinases (SsChi50) Extracted from Offal Red Scorpion Fish in Biological Control of Chickpea Weevil (C allosobruchus maculatus L.)

Production of ChiA-Pt70, a new organic solvent-tolerant extracellular chitinase from &lt;em&gt;Paenibacillus timonensis&lt;/em&gt; strain LK-DZ15

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Production of ChiA-Pt70, a new organic solvent-tolerant extracellular chitinase from <em>Paenibacillus timonensis</em> strain LK-DZ15