空气负离子对阿维链霉菌发酵周期及产量的影响

为了研究低浓度空气离子对微生物的效应,利用低浓度空气离子对固体培养基上Streptomyces avermitilis作用,分析了其次生代谢产物产生时间早晚,各组分含量变化,并且初步研究有效成分含量提高的机理.用浓度为5×103/cm3的空气负离子连续十天处理固体培养基上的阿维链霉菌,经过高效液相色谱(High performance liquid chromatography,HPLC)检测后发现:经过处理,该菌株提早四天到达阿维菌素(Avermectins,AVM)产素平台期,有效成分B1a和B1b提高约20%,而无效成分含量无显著提高.经过分析,低浓度空气负离子可有效缩短该菌种的发酵周期,提高有效成分含量,刺激次生代谢过程.     

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%.     

Construction and evaluation of a three-dimensional structure of cytochrome P450choP enzyme (CYP105C1)

The purpose of this work was to develop and carefully evaluateimproved strategies for constructing reliable 3-D models ofP450 isozymes. To this end, a unique combination of steps forbuilding and evaluating a model structure was used to builda homology model of the P450choP isozyme, based on knowledgeof the X-ray structures of P450cam, P450terp, P450BM-3 and P450eryF.Specifically, the reliability of this model was examined bysystematic comparisons of its conformational, energetic, environmentaland packing properties and those of the four reference proteinswith corresponding properties from the database of proteinswith known structures. The results showed that the examinedproperties of this model structure are well within the criteriaestablished for reliable structures and are of nearly as goodquality as those of the reference proteins. In addition, theresult from a 120 ps unconstrained MD simulation of the modelwith structural waters provided evidence that the model is stableat room temperature. This 3-D model can now be reliably usedfor explicit characterization of substrate and inhibitor complexes.Most importantly, although it is envisioned that building modelsfor mammalian P450s will be even more challenging, the stepsdescribed here should be very useful in future constructionof 3-D models of mammalian P450 isozymes.     

Stabilized variant of Streptomyces subtilisin inhibitor and its use in stabilizing subtilisin BPN'

Protein protease inhibitors could potentially be used to stabilize proteases in commercial products such as liquid laundry detergents. However, many protein protease inhibitors are susceptible to hydrolysis inflicted by the protease. We have engineered Streptomyces subtilisin inhibitor (SSI) to resist proteolysis by adding an interchain disulfide bond and removing a subtilisin cleavage site at leucine 63. When these stabilizing changes were combined with changes to optimize the affinity for subtilisin, the resulting inhibitor provided complete protease stability for at least 5 months at 31 degrees C in a subtilisin-containing liquid laundry detergent and allowed full recovery of the subtilisin activity upon the dilution that occurs in a North American washing machine.     

Optimization of nitrilase production from a new thermophilic isolate

A new thermostable nitrilase‐producing isolate identified as Streptomyces sp. MTCC 7546 has been studied extensively for the optimization of enzyme production operating in batch mode. The benzonitrile was observed as inducer of nitrilase production. The isolate showed maximum nitrilase production after 24 h of incubation at optimal conditions. The strain grows well on a variety of carbon sources and produces the nitrilase that catalyses the hydrolysis of nitriles to acids without formation of amides. The enzyme is mostly active against mono‐ and di‐aliphatic nitriles (10 mmol L^?1) at pH of 7.4 and at a temperature of 50 °C. Copyright © 2007 Society of Chemical Industry     

微生物谷氨酰胺转胺酶的分批发酵生产

在首先采用我们自行设计的2L小型、便易的生化反应器,对我们研究室保藏的高产酶菌株链霉菌(Streptomyces sp.)WZFF.L-M168发酵生产微生物谷氨酰胺转胺酶(MTG)过程中发酵培养基的碳源、氮源和初始pH值的影响作用进行研究,确定培养条件后,逐级扩大发酵罐规模,在20L和200L发酵罐上以在线监控技术手段直接监测分析环境因素对MTG发酵生产的作用效果,确立各级发酵罐分批发酵的生产工艺条件。结果表明:碳氮源采用葡萄糖、淀粉和多价胨,初始pH值为7.0,接种量5%～10%,发酵过程中在线控制培养温度、pH、通气量和搅拌速度等各项参数指标分别为30±0.5℃、6.6～6.9,0.8～1.1vvm和200～400r/min较为适宜。在这优化技术条件下,200L发酵罐可以稳定生产酶活在2.75U/ml以上的MTG。     

Development of an Intergeneric Conjugal Transfer System for Xinaomycins-Producing Streptomyces noursei Xinao-4

To introduce DNA into Streptomyces noursei xinao-4, which produces xinaomycins, we explored an intergeneric conjugal transfer system. High efficiency of conjugation (8 × 10⁻³ exconjugants per recipient) was obtained when spores of S. noursei xinao-4 were heat-shocked at 50 °C for 10 min, mixed with Escherichia coli ET12567 (pUZ8002/pSET152) in the ratio of 1:100, plated on 2CMY medium containing 40 mmol/L MgCl₂, and incubated at 30 °C for 22 h. With this protocol, the plasmids pKC1139 and pSET152 were successfully transferred from E. coli ET12567 (pUZ8002) with different frequencies. Among all parameters, the ratio of donor to recipient cell number had the strongest effect on the transformation efficiency. In order to validate the above intergeneric conjugal transfer system, a glycosyltransferase gene was cloned and efficiently knocked out in S. noursei xinao-4 using pSG5-based plasmid pKC1139.