Induction And Identification Of Pseudomonas Aeruginosa NY3 Alkanes Oxidase

Oil pollution problem is a worldwide problem, enzymes that catalyze the terminal or subterminal oxygenation of alkanes is a hotspot in the research area.Many microbes can with long-chain alkanes as the substrate growth, but growth process produces oxidase and its expression less condition.This paper used pseudomonas aeruginosa NY3 as the main research object, which isolated by the laboratory from oil contaminated soil.First, we inspects the diversity of available carbon source and the oxidase activity, at the same time, inspected the differences of alkane oxidase activity induced by various carbon source, alkanes oxidase of this strain also was made a preliminary identification. By experimental studies we got the following results:(1) Pseudomonas aeruginosa NY3 is able to grow on various carbon source, which can induce the alkane oxidase, including glucose, fructose, saccharose and lactose etc sugar, C5-C16 and diesel, l-butanol, pentanol,1-hexanol, octanol, etc and some organic acid as the only carbon source and energy to grow.When glycerine and hexadecylalcohol as carbon source, the enzyme activity was highest, the degradation rate of substrate was 39.05% and 38.78% ,respectively after 30min.(2) Research results show that NY3 could better applied to the degradation of long chain hydrocarbon. When octane was degraded, added ammonium acetate and lactic acid as start-up factors could improve the enzyme activity. Regard to hexadecane, added ammonium acetate as start-up factor could improve the enzyme activity 29.86%.(3)Iron ions and copper ions have certain effect on the expression of alkane oxidase activity, preliminary identified pseudomonas aeruginosa NY3 alkanes oxidase have similar the sMMO and pMMO enzyme. The expression in the alkane by the Cu²⁺ regulation; Medium copper ion concentration≤50μM, sMMO expression; When copper ions concentration > 50μM coefficient sMMO closed, pMMO expression. The research results of this thesis for in-depth study of alkane oxidase and the application of NY3 as a microbial remediation on oil pollution environment laid a foundation.