CONSTRUCTION OF BACTERA THAT DEGRADE POLYCYCLIC AROMATOC HYDROCABONS AND THEIR APLICATION TO BIOREMEDIATION FOR POLYCYCLIC AROMATOC HYDROCABONS CONTAMINATED SOIL

1)对粪产碱菌AFK2菲代谢操纵子(PHN)的核苷酸序列进行了克隆和序列分析。PHN操纵子由24kb长的13个顺反子组成。2)克隆并分析了恶臭假单胞菌OUS82(Partiomonas putida OUS82，PAH)中一种抑制萘分解操纵子(Pah)的调节子ACC。Acc基因由780个核苷酸组成，编码一个由259个氨基酸组成的蛋白质，分子质量为30K。与通常的阻遏因子不同，acc不与pah操纵子控制区结合。琥珀酸可诱导Ace的产生。3)从土壤中分离到两株降解二苯并呋喃和二苯并-对二恶英的菌株HKT9991和TKT9992。它们分别被鉴定为单纯诺卡氏菌和脱氮产碱杆菌。它们降解了联苯、二苯并噻吩苯和菲。在污染土壤中加入脱氮曲霉TKT9991细胞，可促进二苯并对二恶英的降解。Pickettii Ralstia ickettii DTP0602对4-氯辛醇具有较强的降解能力，可用于二恶英污染土壤的修复。

1) Cloning and sequence analysis of the nucleotides of the phenanthrene metabolic operon (phn) of Alcaligenes faecalis AFK2 were carried out. The phn operon consisted of thirteen cistrons in a 24kb-long region. Similarities the phn in nucleotides sequence were mapped between the naphthalene and biphenyl ectabolic operon operons of Pseudomonas strains.2) A regulator, Acc, that repressed erpression of the naphthalene catabolic opcon of Pseudomonas putida OUS82 (pah) was cloned and analyzed. The acc gene consisted of 780 nucleotides and encoded a protein consisting of 259 amino acids with a molecular mass 30K.The Acc did not bind to the control region of the pah operon unlike an usual repressor. Production of Ace was induced by the succinate. Catabolite repression to the naphthalene catabolism was derepressed and salicylate catabolism was repressed in an acc mutant strain.3) Two species of bacteria, HKT9991 and TKT9992, which degraded dibenzofuran and dibenzo-p-dioxin were isolated from soil. They Were identified as Nocardioides simplex and Alcaligenes denitrificans, respectively. They degraded biphenyls, dibenzothiophene, and anthracene. Addition of cells of A.denitrificans TKT9991 to the contaminated soil facilitated decreased of dibenzo-p-dioxin. Ralstonia pickettii DTP0602 was shown to be a powerful degrader of 4-chlorocatecol and also suggested useful for remediation of dioxin-polluted soil.