两性霉素B合成基因簇及其组合生物学进展

 两性霉素B（AmB）作为第一个用于深部真菌感染的药物,自上市以来已使用超过半个世纪,目前仍是临床治疗中不可缺少的药物。随着近年来AmB衍生物药物的发展,以及其生产菌株结节链霉菌（Streptomyces nodosus）全基因组和AmB体内合成代谢途径的解析,运用基因工程和代谢工程的方法对AmB组合生物合成的研究逐渐增多。由于AmB在当今临床治疗上的地位仍极其重要,因此促进AmB产业的发展具有显著的经济及社会效益。本文对AmB进行了概述,并对其合成基因簇相关基因及其当前组合生物合成的研究现况进行了总结与探讨。     

癸酸抗性突变株流加发酵法生产达托霉素

采用He-Ne激光对达托霉素产生菌玫瑰孢链霉菌LC-54进行诱变,筛选前体癸酸抗性突变株,在此基础上进一步考察了一次补料、间歇补料和恒速流加补料等不同补料策略对达托霉素发酵生产的影响．结果表明,选育前体癸酸抗性突变株可以提高达托霉素的生产能力,并最终获得了达托霉素发酵效价较出发菌株提高了37．2%的高产突变株LC-KS-54．采用恒速流加策略,能够有效地提高达托霉素产量,经过130 h的恒速流加培养,细胞干重达到15．4g/L,达托霉素产量达到258 mg/L,明显优于其他前体补料策略．     

Enhancement and selective production of avermectin B by recombinants of Streptomyces avermitilis via intraspecific protoplast fusion

Among eight components of avermectin, B1 fractions have the most effective antiparasitic activities and the lowest level of toxic side-effects and are used widely in veterinary and agricultural fields. In-traspecific protoplast fusion between two strains of Streptomyces avermitilis, one an avermectin high producer (strain 76-05) and the other a genetically engineered strain containing the mutations aveDˉ and olmAˉ (strain 73-12) was performed for enhancement and selective production of avermectin B in the absence of oligomycin. Two recombinant strains (F23 and F29) were isolated and characterized with regards to the parental merits. F23 and F29 produced only the four avermectin B components with high yield and produced no oligomycin. The avermectin production of F23 and F29 was about 84.20% and 103.45% of the parental strain 76-05, respectively, and increased about 2.66-fold and 3.50-fold, re-spectively, compared to that of parental strain 73-12. F23 and F29 were genetically stable prototrophic recombinants and F29 was quite tolerant of fermentation conditions compared to avermectin high producer parental strain 76-05. The ability to produce avermectin B with high yield without the produc-tion of other avermectin components and oligomycin will make F23 and F29 useful strains for aver-mectin production. Strain F29's tolerance of fermentation conditions will also make it suitable for in-dustrial applications.     

链霉菌属一新种的多相分类

 从云南西双版纳分离到的1株中温放线菌N-98-1272(以下简称1272).对其形态,生理生化特性,细胞壁化学组分以及16SrDNA序列进行了研究分析,并与链霉菌属的已知种进行了比较,认为这是链霉菌属的1个新种,命名为版纳链霉菌(Streptomyces bananensis sp.nov.).     

利用紫外线诱变原生质体筛选四环素高产菌株的研究

以金色链霉菌为出发菌株，经溶菌酶作用制备原生质体，然后以紫外线对原生质体进行诱变处理，在再生培养基上再生培养，获得再生突变株，将其中的单菌落以琼脂挖块法进行初筛获得第２０号菌株，其相对效价为１．２８４。     

克拉维酸高产菌的选育

在克拉维酸高产菌的选育过程中,应用解除终产物结构类似物作为选择压力,能有效地除去低效价菌株,提高高产菌株的检出率,较传统筛选法优越.试验表明,以带棒链霉菌Strep tomycesclavuligerusCCRC11518(效价为382.4×10-6g/mL)为出发菌,经紫外线诱变,筛选到一株抗20×10-3g/mL的舒巴坦钠突变株my51,my51菌株产克拉维酸的效价达到834.7×10-6g/mL,比出发菌提高了1.18倍.该菌株连续传代7代,克拉维酸的效价保持稳定.     

阿维链霉菌中转录因子AveR对阿维菌素生物合成的正调节

aveR是阿维链霉菌NRRL 8165阿维菌素生物合成基因簇中唯一可能的调节基因.为了验证aveR是否参与阿维菌素生物合成基因的转录调节,构建了用于敲除aveR的基因置换质粒pJTU2530,并通过接合转移引入了阿维链霉菌.通过筛选ThioSAprR转化子,经聚合酶链式反应(PCR)扩增验证,获得了aveR内部1 320 bp区域被阿泊拉霉素抗性基因aac(3)IV替换的突变株ZD10.高压液相色谱检测表明,与野生型菌株相比,突变株ZD10不再产生阿维菌素,并且ZD10中寡霉素的产量明显高于野生型菌株.进一步的反转录PCR(RT-PCR)分析表明,与野生型菌株相比,突变株ZD10的聚酮合酶基因aveA3不再转录.结果显示,AveR是阿维菌素生物合成的正调节因子,通过调节结构基因的转录表达来影响阿维菌素的产生.     

Properties and characterization of Au3+-adsorption by mycelial waste ofStreptomyces aureofaciences

Mycelial waste of Streptomyces aureofaciences procured from the aureomycin fermentation industry is used as biosorbent for Au³⁺  The properties of Au³⁺ adsorption by the mycelial waste are studied. The results indicate that the optimum pH value of Au³⁺ adsorption is 3.5. The biosorption is a rapid and non-temperature-dependent process. The bio-sorptive capacity with 45.6 mg/g and efficiency with 91.2% are achieved under the conditions of pH 3.5 and 30 ℃ for 45 min, in which the ratio is 50 mg/g dry weight for the concentrations of initial Au³⁺ and the mycelial waste. The Au³⁺ ions adsorbed on the mycelial waste can be eluted. The observation in a transmission electron microscope shows that the Au³⁺ ions can be reduced to Au particles by the mycelial waste and the Au⁰ can become gold crystals with differentforms and sizes. X-ray photoelectron spectroscopy analysis further proves that the Au³⁺ can be reduced to Au⁰ by the mycelial waste.     

链霉菌G-HD-4产黑色素的提取及理化性质

对链霉菌G-HD-4发酵液采用酸沉淀、碱性溶液提取、酸水解,以及多种有机溶剂抽提等工艺手段分离、纯化出黑色素,研究其理化性质.结果表明,该黑色素的最大吸收峰为246 nm,与标准黑色素最大吸收峰相一致.在400 nm波长下,其光密度值(D)随黑色素质量浓度的增加而增大;容易被强氧化剂所氧化,而紫外、光照、温度、有机溶剂、还原剂、各种盐,以及金属离子对黑色素的影响较小,氨基酸、淀粉等添加物对黑色素稳定性没有影响,说明该黑色素稳定性良好.     

链霉菌酯酶研究进展

酯酶是一类催化酯键水解和形成的酶类,主要包括脂肪酶和羧酸酯酶.作为生物催化剂,酯酶具有重要的应用价值并被广泛应用于工业领域.链霉菌富含丰富的、生物学特性多样的酯酶.本文对链霉菌酯酶的表达及分泌调节、酶学性质及晶体结构、筛选及发酵生产三个方面进行了综述,以期引起人们对于链霉菌酯酶研究的兴趣.