Tenebrio antifreeze proteins

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/12 (2006.01) A01K 67/00 (2006.01) A01K 67/027 (2006.01) C07K 14/435 (2006.01) C07K 16/18 (2006.01) C09K 5/00 (2006.01) C12N 1/15 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 15/70 (2006.01)

Patent

CA 2296005

Thermal hysteresis (antifreeze) proteins (THP) that have up to 100 times the specific activity of fish antifreeze proteins have been isolated and purified from the mealworm beetle, Tenebrio molitor. Internal sequencing of the proteins, leading to cDNA cloning and production of the protein in bacteria has confirmed the identity and activity of the 8.4 to 10.7 kDa THP. They are Thr- and Cys-rich proteins composed largely of 12-amino-acid repeats of cys- thr-xaa-ser-xaa-xaa-cys-xaa-xaa-ala-xaa-thr. At a concentration of 55 micrograms/mL, the THP depressed the freezing point 1.6 ~C below the melting point, and at a concentration of 1 mg/mL the THP or its variants can account for the 5.5 ~C of thermal hysteresis found in Tenebrio larvae. The THP function by absorption-inhibition mechanism and produce oval-shaped ice crystals with curved prism faces.

Protéines à hystérèse thermique (antigel) (THP), possédant jusqu'à 100 fois l'activité spécifique de protéines antigel de poisson, qui ont été isolées et purifiées du ténébrion meunier (Tenebrio molitor). Le séquençage interne des protéines, conduisant au clonage d'ADNc et à la production de la protéine dans des bactéries, a confirmé l'identité et l'activité de la THP de poids atomique entre 8,4 à 10,7 kDa. Il s'agit de protéines riches en Thr et en Cys composées principalement de répétitions des 12 acides aminés cys-thr-xaa-ser-xaa-xaa-cys-xaa-xaa-ala-xaa-thr. A une concentration de 55 microgrammes/ml, la THP abaisse le point de congélation de 1,6 ·C au-dessous du point de fusion, et à une concentration d'environ 1 mg/ml, la THP ou ses variants peuvent être à l'origine des 5,5 ·C d'hystérèse thermique que l'on trouve dans les larves de ténébrion. La THP fonctionne par un mécanisme d'adsorption/inhibition et produit des cristaux de glace de forme ovale à faces prismatiques incurvées.

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