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Screening and Characterization of Probiotic Lactobacillus Strains with Inhibitory Activity to Monoamine Oxidase

Author: WangHao
Tutor: GuoBenHeng
School: Shanghai Ocean University,
Course: Of Food Science
Keywords: Anti-Aging Probiotics Monoamine oxidase Filter Inhibitory activity
CLC: TQ460.1
Type: Master's thesis
Year: 2009
Downloads: 100
Quote: 0
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Abstract

Population aging in the 21st century, mankind faces a major challenge. Age-related diseases, such as Alzheimer's disease, Parkinson's disease, etc., has become enemy of human health and social prosperity of the heavy burden. At present, with the aging population amplification, can provide prebiotic role in controlling aging and prolong the life of functional foods more and more popular. Lactic acid bacteria, especially Lactobacillus as a class recognized as safe mushroom (generally regarded as safe, GRAS), on human health regulation and catalytic role in constantly being discovered and confirmed its potential anti-aging effects gradually aroused widespread concern. In recent years, about probiotics lowering blood pressure, anti-hypertensive, cholesterol, etc. to improve cardiovascular health function has been confirmed, but for probiotic anti-aging effect of very little. In fact, back in 1905, the famous microbiologist Meiqi Kafelnikov discovered Caucasus region residents to drink lactic acid produced yogurt can live long, but now probiotic anti-aging research is still trying to use an animal model for intervention experiments, yet See a probiotic in vitro anti-aging research reports, this may be due to aging mechanisms of physiological and biochemical changes is more complex, so the in vitro anti-aging function evaluation lack of simple aging indicators. Monoamine oxidase (MAO) The main function is to catalyze endogenous and exogenous monoamines metabolism in the human body is considered to be the signs of aging, also known as age-related, the enzyme activity rising cause the entire body functions, especially brain function gradually degradation, aging. In this study MAO as a target enzyme, the establishment screened for inhibition of monoamine oxidase probiotics in vitro model, for the further in vivo evaluation of probiotics anti-aging effect reference. MAO inhibitors means that there is a potential anti-aging effects, this paper in vitro inhibition of monoamine oxidase probiotic active ingredients were screened. By acetone extraction techniques and thin layer chromatography (TLC) on the strain cell contents for the extraction and initial separation, determination of the different fractions of the MAO inhibitory activity, select the effect on the inhibition of the component to the national drug Screening Center for further evaluation. At last, the isolates were related to the physiological function and growth characteristics studied by molecular biology methods and API carbohydrate fermentation test for screening isolates were identified. Screening experiments strain JH-23 cell lysate of MAO showed a strong inhibitory activity screening inhibition rate was 33.7%, the reaction concentration of 16mg/mL inhibition rate can be increased to 53.2%. Molecular retention capacity of 1 KD dialysis bag of the cell lysate was dialyzed found that the main active ingredient transferred to the dialysate, the description play an inhibitory composition of the bacterial cells of the small molecule product (MW lt; 1 KD). Therefore, the use of acetone extraction method for bacterial cells having MAO inhibitory small molecules extracted. Thin layer chromatography (TLC) of JH-23 intracellular crude extract of the extraction and separation, and ultimately get four components, wherein component Ⅰ on MAO inhibition of the most significant, the reaction concentration of 120μg/mL the MAO inhibition rate reached 55.4%, and there is significant dose effect. The ingredients Ⅰ enrichment freeze-dried and then sent to National Center for Drug Screening Department to conduct MAO activity assay, confirmatory component Ⅰ on MAO inhibition. Strain JH-23 anaerobic growth, Gram-positive, cell rod, non-spore. The 16S rDNA sequence alignment and API carbohydrate fermentation experiments identified as Lactobacillus casei (Lactobacillus casei), named Lactobacillus casei JH-23. The Lactobacillus has a strong acid, bile tolerance and NaCl tolerance, its optimum growth temperature is 37 ℃, the optimum pH was 6.0, in MRS medium vigorous growth, skim milk growth is slow and not curd.

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