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Study on Enzymological Characterization and Anti-cancer Effect of Tenebrio Fibrinolyric Proteins

Author: WuYanLing
Tutor: TanZhuJun
School: Guangdong University of Technology
Course: Biochemical Engineering
Keywords: Tenebrio Fibrinolytic activity of protein Enzymatic Properties Michaelis-Menten constant (Km) Serine protease Tumor suppressor role
CLC: S899.9
Type: Master's thesis
Year: 2011
Downloads: 26
Quote: 0
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Tenebrio rich in protein and other nutrients, is a valuable edible insects resources, has been widely used in human food, health care products, pharmaceuticals and feed. The Tenebrio viability and ability to adapt to rapid growth and development, farming and low cost, has become the world today after the sericulture the beekeeping development of one of the fastest insect rearing industry. Has proven from the the Tenebrio body extracted an active protein, with a good thrombolytic effect, cancer and cardiovascular disease is the world's two major killer of life, hope that further study of the active protein, to provide a more complete basis of experimental data lay the foundation for the future to find an efficient, safe, low-cost drugs. This article is homogenized extraction, salting, anion exchange, and dextran gel filtration method, Tenebrio larvae of extraction and purification of a fibrinolytic activity protein (Tenebrio Fibrinolyric Proteins TFP) fiber plate method good fibrinolytic effect, non-denaturing electrophoresis to prove its purity, as a single component, denaturing electrophoresis measured relative molecular weight of about 56.1kD protein concentration of 6.061mg · mL-1. Learn more about this enzyme enzymatic properties by TFP role on three kinds of substrates (BAEE, BAPNA and casein), measured Michaelis constant Km values, which BAEE as substrate Km value of 0.182 mmol / L could not be determined, significantly less than BAPNA as substrate Km value (1.138 mmol / L), and casein as substrate Km values. The BAEE ester bond breakage TFP acting on BAEE exhibit esterase activity; acting on BAPNA BAPNA amide bond cleavage, the performance of the enzyme activity of non-peptide amide bond, an amide; acting on casein and fibers protein cleavage of the peptide bond, peptide bond of enzyme activity, so draw: TFP esterase activity gt; lactamase vitality gt; enzyme activity of the peptide bond. Simulation environment of the small intestine, alkaline, TFP is not trypsin degradation, enteric preparations can be done. To explore the the enzymology types of TFP, the serine protease inhibitor PMSF role in TFP, fiber plate method measuring inhibition of enzyme activity, the results show that the PMSF concentration of 5mmol / L can be completely inhibited by TFP fibrinolytic activity, indicating that PMSF TFP can effectively inhibit fibrinolysis activity, the inhibition of TFP to its concentration was positively correlated. According to the results of infrared spectroscopy, that trypsin and TFP structure very close, trypsin is a serine protease, protease active site on the phylogenetic has a strict conservative, and thus concluded that TFP is a serine protease. The article also try the anti-cancer effect of TFP made a preliminary study by in vitro culture of animal cancer cells, MTT assay, the results showed significant inhibition of Tenebrio fibrinolytic activity of protein on the tumor cells, A549 cells, H22 The cell inhibition obvious selective for cancer cells. In addition, TFP and anti-cancer drug cisplatin, the effect of the tumor cell line A549, derived: TFP anticancer effect very quickly, a short period of time can achieve better effect of inhibition of tumor growth, which is unparalleled of other anti-cancer drugs advantages. May they not the same mechanism of action remains to be further research.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Beekeeping,beneficial insects feeding > Beneficial insects feeding > Other
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