一株转化反式茴香脑产茴香酸菌株的筛选鉴定

摘要

After the gradient acclimation and enrichment screening,348 strains were isolated from environmental samples using trans-anethole as the sole carbon source.Through quantitative detection of trans-anethole and anisic acid with the methods of TLC and RP-HPLC,a strain named WGBF9 with the highest tolerance to trans-anethole and maximum yield of anisic acid was obtained.Biomass of strain WGBF9 was 37.42％ of the blank control when the seed culture medium contained 10％ (v/v) transanethole,and the biomass reached 26.57％ in the medium with 20％ (v/v) trans-anethole,showing that the strain had a high tolerance to the trans-anethole.Anisic acid molar generation rate is 17.9％,when the strain was cultured in the screening secondary medium,with rotate speed of 200 r/min,after 36 h fermentation under 30 ℃.WGBF9 was identified as a subspecies of Pseudomonas putida biotype A based on the analysis of physiological and biochemical experiments.%经过梯度驯化及富集筛选,从环境样品中分离获得348株能以反式茴香脑为唯一碳源的菌株.通过薄层层析法和反相高效液相法对转化液中的残余反式茴香脑和茴香酸进行定量分析,最后得到一株对反式茴香脑耐受能力和茴香酸产量都较高菌株WGBF9.在含体积分数为10％的反式茴香脑的种子培养基中,菌体的生物量为空白对照的37.42％,在含体积分数为20％反式茴香脑时,生物量也能达到空白对照的26.57％,说明WGBF9对反式茴香脑具有很高的耐受能力.将该菌株接种到复筛培养基中,30℃、转速200 r/min的条件下发酵36 h,茴香酸的摩尔生成率为17.9％.经形态学观察、生理生化等实验分析,初步鉴定WGBF9为恶臭假单胞菌A类变种(Pseudomonas putida biotype A)中的一个亚种.