Studies On A-pinene Metabolic Flux Analysis As Well As Molecular Biology In Pseudomonas Veronii ZW

α-pinene is a typical Volatile Organic Compounds, it is harmful for human body and environment. Microbial technique is a useful methods for α-pinene degradation. Through the analysis of protein expression stressed with pressure can identify the enzymes which participate in metabolize of environmental pollutants. It is provide evidence of process of metabolize of environmental pollutants by microorganism.An approach combining traditional biological methods and modern molecular biological techniques was used for investigating the stress response of strain Pseudomonas veronii ZW upon the a-pinene challenge.The experimental results showed that the Haldane modification of the Monod equation adequately described the relationship between specific growth rate and substrate concentration, The maximum specific growth rate and specific degradation rate for α-pinene were0.10253h-1and0.33586h-1, respectively. There are three possible metabolic pathways for the degradation of α-pinene by strain ZW which was detected by GC-MS combined with literatures②α-pinene was converted to myrtenol, furthermore, the myrtenol was converted to myrtle acid;②α-pinene was converted to limonene directly;③α-pinene was converted to p-cymene, then the p-cymene was converted to p-cumic alcohol and3-hydroxy-4-isopropyl toluene. At last, the p-cumic alcohol would form low molecular weight organic acids like2-methyl-1,4-butanedioic acid, oxalic acid, acetic acid enter into TAC cycle.Using UV mutagenesis obtained a strain ZW-U which loss the α-pinene degradation ability, its morphology and antibiotic susceptibility basically consistent with the normal strain ZW. The degradation ability of mutant strain ZW-U for a-pinene, p-cymene, p-cumic alcohol,2-methl-1,4-butanediol decreased and even lost, it may be related to the strain ZW-U through UV mutagenesis that loss the degrading enzyme for α-pinene, p-cymene, p-cumic alcohol,2-methl-1,4-butanediol, or inhibited synthesis of these enzymes.16kinds of specific proteins was isolated and identified by2-DE and MALDI-TOF-MS. Proteins were grouped in6different functional families, taking into consideration only one of the protein biological roles. A series of proteins which related to amino acid biosynthesis, energy conversion, protein folding, detoxification, protein regulation were induced in response to α-pinene.The identification of PKHD-type hydroxylase and aryl-alcohol dehydrogenase related to the degradation of α-pinene by Pseudomonas veronii ZW. PKHD-type Hydroxylase is probably transforme cymene into3-hydroxy-4-isopropyl toluene. Aryl-alcohol dehydrogenase is probably converte p-cumic alcohol to p-cumic aldehyle.