蓝莓溃疡病生防链霉菌CX3的抑菌效果及其鉴定

[目的]为了明确从吉林省长白山保护区池西保护站马鞍山云杉人工林林下土壤分离,所获得1株拮抗菌株(编号记为CX3)的生防效果及分类地位。[方法]分别采用平板对峙法和杯碟法测定该菌株活菌及其发酵液的抑菌活性和抑菌谱;并通过形态学、生理生化特征、16S rRNA序列分析等方法研究了其分类地位。[结果]该菌株对13种供试植物病原菌均有抑制作用,抑菌谱广,其中对蓝莓溃疡病菌有极强的抑制作用,抑菌带达25.91 mm,发酵液的抑菌圈直径达31.73 mm;根据其菌株形态、生理生化特性、16S rRNA序列比对,最终鉴定菌株为链霉菌属Streptomyces sp.。[结论]该菌株对蓝莓溃疡病菌有显著的抑制效果,应用开发前景良好,此次是国内首次报道该菌株杀真菌活性。     

应用基因组重排技术提高普那霉素产量

将普那霉素产生菌——始旋链霉菌ND-23的孢子和原生质体经紫外线诱变后获得高产突变菌株,其中高产突变株SP-S73的产量达到356 mg/L,比出发菌株提高了31%.在上述增产突变株中选取4株作为基因组重排的出发亲本,对它们进行了4轮基因组重排育种,筛选得到了高产重排菌株,其中1株重排菌株G-211的普那霉素产量为832 mg/L,比产量最高的亲本菌株提高了134%,比原始出发菌株ND-23(272 mg/L)提高了206%.通过研究高产菌株和出发菌株ND-23在5 L罐上的发酵过程,发现普那霉素产生菌的生物合成属于非生长耦合型;其高产菌株G-211在合成产物的时期对还原糖和氨基氮的消耗量均大于原始菌株ND-23,这些结果将为高产菌株发酵条件优化和发酵放大研究提供有价值的参考.     

去甲基金霉素发酵培养基优化研究

为提高金霉素链霉菌生产去甲基金霉素的效价，通过正交试验研究了发酵培养基中的碳源、氮源、无机离子的影响，求得较优的培养基为淀粉６％、蛋白胨１％、黄豆饼粉５％、玉米粉２％、酵母粉０．４％、ＮＨ４Ｃｌ０．１６％、ＣａＣＯ３１％、豆油１％、α－淀粉酶０．０３％，其效价达到３４９３，比原培养基提高３９％。     

链霉菌TD-1菌株抑菌活性物质发酵条件的优化

利用正交设计法及均匀设计法对链霉菌TD-1菌株发酵条件进行了优化,研究不同发酵因子对链霉菌TD-1产抑菌活性物质的影响.结果表明,发酵培养基中影响抑菌活性大小的主要因素为葡萄糖、黄豆粉、硫酸亚铁、磷酸氢二钾及硝酸钾;抑菌活性物质较佳发酵条件为90 mL/250mL三角瓶,接种量的体积分数为0.06,发酵周期8 d,初始pH值为7.15.在此条件下,最终抑菌效率与原发酵液相比发酵液对串珠镰刀菌抑菌活性提高了53.38%.     

Interrogating the Tailoring Steps of Pactamycin Biosynthesis and Accessing New Pactamycin Analogues

Pactamycin is a bacteria‐derived aminocyclitol antibiotic with a wide‐range of biological activity. Its chemical structure and potent biological activities have made it an interesting lead compound for drug discovery and development. Despite its unusual chemical structure, many aspects of its formation in nature remain elusive. Using a combination of genetic inactivation and metabolic analysis, we investigated the tailoring processes of pactamycin biosynthesis in Streptomyces pactum. The results provide insights into the sequence of events during the tailoring steps of pactamycin biosynthesis and explain the unusual production of various pactamycin analogues by S. pactum mutants. We also identified two new pactamycin analogues that have better selectivity indexes than pactamycin against malarial parasites.     

A Taguchi design approach for the enhancement of a detergent-biocompatible alkaline thermostable protease production by Streptomyces mutabilis strain TN-X30

The ability of microorganisms to grow at high temperature, alkaline pH, and high salinity makes them an attractive target for enzyme-production with several industrial applications. One strain TN-X30 has been selected as protease producer and identified as Streptomyces mutabilis after a phenotypic and molecular study. Its production of protease was improved using Taguchi L27 design. The strategy was carried out to identify the optimum levels and the interaction of the screened factors. Following this step, maximum protease activity (10,895 U/ml) was achieved after 6-days of incubation. The TN-X30 protease activity had an optimum of pH and temperature of 10 and 65°C, respectively. Thermodynamic parameters at 60°C were enthalpy 14.26 kJ/mol, entropy −220 J/mol/K, and Gibbs free energy 90.53 kJ/mol. TN-X30 protease production displayed a 16-fold increase reaching 175,000 U/ml in a 100-L fermentor. Furthermore, the lyophilization in presence of sorbitol enhanced the stability of the TN-X30 protease which remained active at 75% after 24-months of storage. The lyophilized TN-X30 protease exhibited exceptional stability indexes in presence of some known commercialized detergent components as NEODOL® 25-7, Dehydol® LT 7, Na₂ CMC, Galaxy LAS, Galaxy LES 70, Galaxy 110, Galaxy CAPB Plus, and Sulfacid K. The lyophilized enzyme also displayed high stability with respect to both solid and liquid detergents. Finally, TN-X30 protease exhibited remarkable destaining of blood, egg, and chocolate stained cloth pieces. These findings may promote TN-X30 protease for use as bioadditive in detergent formulation, thereby reducing environmental chemical threat.     

Biosynthetic approach for the production of new aminoglycoside derivative

Aminoglycoside antibiotics can be classified into two major groups; streptamine containing and 2-deoxystreptamine containing antibiotics. Here, we report a biosynthetic approach for the fusion of spectinomycin and kanamycin biosynthetic gene clusters to yield the new aminoglycoside derivative, oxykanamycinC, in a non-aminoglycoside producing heterologous host.     

双丙氨磷发酵过程变温控制的研究

为了考察不同温度对双丙氨磷的发酵影响,采用摇瓶发酵,改变发酵温度,来确定较为适合的发酵工艺。结果表明,双丙氨磷吸湿链霉菌的菌丝最佳生长温度是28℃,产物的最佳合成温度是26℃。采用变温控制方案,双丙氨磷终产物比恒温培养提高了11.7%。     

培养条件对褐黄孢链霉菌发酵合成纳他霉素的影响

对褐黄孢链霉菌摇瓶发酵合成纳他霉素的研究表明: 种龄、接种量和装液量对纳他霉素合成的影响显著;在实验范围内,随着种龄、接种量和装液量的增加,纳他霉素产量均表现出先增加后减少的趋势,而菌体干重则变化很小.最适宜产物合成和菌体生长的发酵培养基初始pH值各不相同,分别为7.5～8.0和6.5～7.0.当发酵温度从31℃下降到25℃时,纳他霉素产量大幅度增加,而菌体干重在31℃达到最大值.在种龄48 h、接种量10%、25℃、初始pH值 7.6、装液量10%的优化条件下,纳他霉素最高产量达到1.5 g/L, 基于菌体干重的产率为4.917%.