The Biodegrable Of Natural P-Anisaldehyde From Anise-Oil And Establish Analytical Methods For Detecting The Production And Substrate In Bioconversion

In the dissertation, it has studied the process of anethole biodegradation and analyzed the product of biological conversion system. Microbial strains were achieved through appropriate screening method, and anethole was transformed into p-anisaldehyde via selective biodegradable approach. Herein, this method provided a theoretical basis for the future research of biological synthesis of p-anisaldehyde.The main achivements on the preparation of biological p-anisaldehyde are as follows:(1) TLC was carried out for analysis of anethole,p-anisaldehyde and anisic acid. The developing solution of petroleum ether: chloroform: ethyl acetate: formic acid=25:10:3:0.2(v/v/v/v). The method can qualitatively detect these three compounds as soon as possible;(2) The method for simultaneous analysis of anethole,p-anisaldehyde and anisic acid in bioconversion broth has been developed. The chromatographic conditions was 30:70 mixture of 0.02 % acetic acid aqueous solution and acetonitrile as mobile phase, the UV detection wavelength was 260 nm and at a flow rate of 0.8 ml/min. The method can quantitatively analyse the three compounds accurately;(3) A method for the determination of p-anisaldehyde and anisic acid by UV spectrophotometry was developed. This method based on the iso-absorptive point UV spectrophotometry to determine the amount of anisaldehyde and anisic acid simultaneously, respectively. The simple linearrelationships of p (anisaldehyde) =A₂₉₂-0.0064/0.0775 and p (anisic acid) =A₂₆₀-A₂₉₂/0.0775 were convenient to calculate the amount of anisaldehyde and anisic acid, respectively;(4) Innovative launched anethole microbial biodegradation screening. Anethole degradation microorganisms has been screened from the fruits branch,leaf of aniseed, waste residue of anise-oil extraction and the soil near aniseed tree and pilot experiment workshop. A strain ZJ-9 producing p-anisaldehyde was isolated from waste residue of anise-oil extraction and was identified as Aspergillus niger. Then optimize the cultivation conditions of the strain ZJ-9 and the optimal conditions were as followed: the inoculationsize 4.0×10⁵ units/100mL, the fermentation temperature 30℃, a 500mL flask containing 100mL medium, the speed 200 r·min^-1 and The pH 6.0. The cultivation time and the fermentation time were both three days; (5)Anethole achieved a high conversion rate of degradation. The degradation rate of anethole was 82.03% and the mole generation rate of anisaldehyde was 4.54% at the optimization of fermentation media;(6)Some analytical methods were established for characterizing the structure of the key intermediate, such as TLC, GC-MS, IR, NMR, column chromatography. The results showed that the compound was anethole diol and it provided reliable basis for degradation pathway.