大肠杆菌棉子糖操纵子a-半乳糖苷酶表达的调节控制

大肠杆菌棉子糖操纵子(raf)位于质粒,其第一结构基因rafA编码的a-半乳糖苷酶为诱导酶。Rat操纵子比乳糖操纵子(lac)或蜜二糖操纵子(mel)对诱导物有更严格的结构特异性。该酶被蜜二糖或棉子糖诱导,也被D-半乳糖微弱诱导,但不受乳糖、PNPG等结构相近糖所诱导。A-半乳糖苷酶的酶诱导形成能力在对数生长末期出现高峰。Rat 操纵子基因结构组成及调节与乳糖操纵子相似。以阻遏物为中介的负调控在raf操纵子调节中起主要作用,同时以环腺苷一代谢降解物基因激活蛋白(cAMP-CAP)为中介的正调控也参与调节。当0．4％葡萄糖加入到其它碳源培养基时,该酶表达水平下降至原活力的1／2—1／3。无论诱导或组成型酶的葡萄糖抑制均未见瞬时抑制。腺苷环化酶(cya)缺失或环腺苷受体蛋白(crP)和cya双缺陷菌株的酶表达则分别下降到原活力的9％和2．5％。Cya突变株或葡萄糖对raf操纵子表达的抑制可被cAMP解除,但cya和crP双缺陷菌株仍有葡萄糖抑制,而且这种抑制不为cAMP抵消,表明通过降低cAMp而影响cAMP-CAP复合体形成还不能解释代谢降解物抑制的全部机制。尚无证据说明吲哚类小分子化合物和低浓度尿素对raf操纵子表达的明显作用。     

大肠杆菌棉子糖操纵子α—半乳糖苷酶表达的调节控制

The alpha-galactosidase, coded for by the first structural gene rafA in the plasmid determined raf operon was an inducible enzyme. In contrast to lac or mel operon, raf operon has more strict structural specificity for inducers. The enzyme can be induced by melibiose and raffinose, or weakly by D-galactose, but not by structurally related sugars such as lactose, PNPG etc.. The alpha-galactosidase forming capacity as function of growth curve reached a single peak at the end of the logarithmic phase of the growth. The structure and regulation of raf operon is similar to those of lac operon. The repressormor-mediated negative control plays a major role in the regulation of raf operon, and cAMP-CAP mediated positive control is also involved in the regulation. When 0.4% glucose was added into the medium with other carbon sources, the expression of the enzyme was repressed by 2-3 fold. Transient catabolite repression has been observed neither in inducible nor constitutive alpha-galactosidase expression. Based on alpha-galactosidase assay, in mutant strains CA8306(cya) and CA8445 (cya, crp) the expression level of raf operon was only 9% and 2.5% of that in wild type strain respectively. The glucose effect or the repression in cya mutant can be abolished by 1-5 mmol cAMP. The constitutive alpha-galactosidase expression in cya and cry double mutant (CA8445) remains repressible by glucose, but irreversible by cAMP, suggesting cAMP-CAP complex is not the exclusive mediator of the catablite repression.(ABSTRACT TRUNCATED AT 250 WORDS)     

假单胞菌S—42对偶氮染料的脱色和降解代谢

Pseudomonas S-42 was capable of decolorizing azo dyes such as Diamira Brilliant Orange RR(DBO-RR), Direct Brown M (DBM), Eriochrome Brown R(EBR) and so on. The cell suspension, cell-free extract and purified enzyme of Pseud. S-42 could decolorize azo dyes under similar conditions: the optimum pH and temperature laid 7.0 and 37 degrees C respectively. The efficiencies of decolorizing of DBO-RR, DBM, EBR by intact cells stood more than 90%. When the cell concentration was 15 mg(wet)/ml and the reaction time was 5 hours, the decolorizing activity for above three azo dyes by intact cells were 1.75, 2.4, 0.95 micrograms dye/mg cell, respectively. Cell-free extract and purified enzyme could well express the decolorizing activity only under the anaerobic condition and added NADH. Purified enzyme belongs to azoreductase, its molecular weight is about 34,000-2000 daltons, and its Vmax and Km for DBO-RR are 13 mumol.mg protein-1.min-1 and 54 mumol/L. The results of the detection of the biodegrading products of DBO-RR by spectrophotometric and NaNO2 reactional methods showed that the biodegradation of azo dyes was initiated by the reduction cleavage of azo bonds. It was hypothesized that biodegrading metabolism pathway of DBO-RR by Pseudomonas S-42.     

质粒pXZ101 45DNA转化棒杆菌原生质体的研究

用SDS处理谷氨酸棒杆菌1014(Corynebacterium glutamicum 1014),获得消除质粒的衍生株1014—6。通过质粒pXZl0145(Cmr)转化1014—6菌株的原生质体,研究了转化最适条件。0．6u／ml青霉素处理对数期菌体可明显提高转化效率。转化促进剂PEG以分子量6000,浓度30％为最佳。转化在37℃水浴进行3min效果最好。转化效率最高可达2×104转化子/μg DNA。质粒pXZ10145电已成功地转入钝齿棒杆菌B9(C．Crenatu B9)。并在新 宿主中基本保持稳定。     

漆酶、铜蓝蛋白的脉冲激光光声法测定

本文采用脉冲激光光声分析法,进行了漆酶和铜蓝蛋白的测定。探讨了入射激光能量。照射时间、温度等因素对漆酶、铜蓝蛋白光声信号强度的影响以及18种金属离子与铜蓝蛋白的作用情况。     

棉立枯丝核菌的dsRNA与致病力的研究

对我国北方棉立枯丝核菌(Rhizoctonia solani)的18个分离物进行了dsRNA的检测和致病力比较,可见：(1)dsRNA以高频度(16／18)存在于各分离中;(2)各分离物的dsRNA有明显不同的电泳图谱,有1—8个片段,分子量自0．94—4．8×106道尔顿;(3)同地区土壤中分离物常有相同的dsRNA片段;(4)以弱致病力分离物BALr-2的[r-32P]ATP末端标记dsRNA为探针与16个分离物dsRNA进行分子杂交,只有3个分离物有强的同源性,Northern转移杂交表明同源核苷酸在1．15×106的片段;(5)变性电泳示丝核菌的dsRNA泳动率有减慢现象,从而提出环状dsRNA的可能。     

节杆菌82菌株降解胆甾醇生成3-氧代-联原胆烷一1，4-二烯-22-酸

从19株有降解胆甾醇能力的微生物中,筛选出一株节秆菌(Arithrobacter)82菌株,它能在含硫酸钴的培养基中转化胆甾醇和积累3-氧代-联原胆烷-1,4-二烯-22酸(BNc)。转化过程中形成的主要中间体为胆甾烯酮(Cholestenone)。降低培养基中葡萄糖的浓度并增加玉米浆的量,可促进胆甾烯酮侧链的降解,有利于BNC的积累。BNC可在酸性溶液中形成结晶并沉淀下来,故适宜于离心收集。用传统的理化方法及光谱分析技术测定了产物的结构及特性。     

从酵母中制备辅酶Ⅰ的简易方法

辅酶Ⅰ(尼克酰胺腺嘌呤二核苷酸,NAD~+)是一种重要的生化试剂,常从面包酵母中提取。前人报道的提取方法,或是银盐沉淀方法,或是层析方法,由于成本昂贵,操作繁复耗时,反应体积大等缺陷,不甚理想。本文报告一种以小体积汞盐试剂分离纯化NAD~+的简易方法。