New pyrazinoquinazoline alkaloids Isolated from a culture of Stenotrophomonas maltophilia QB-77

Two new pyrazinoquinazoline alkaloids, epi-fiscalin D (1) and epi-fiscalin E (2), as well as three known analogues, norquinadoline A (3), quinadoline A (4), and fiscalin C (5), were isolated from ethyl acetate extract of the fermentation broth of Stentrophomonas maltophilia QB-77. The structures of new compounds were elucidated on the basis of extensive spectroscopic data analysis including UV, HRESIMS, and 1D and 2D NMR experiments. All the isolated compounds were tested for their in vitro cytotoxicity against five human cancer cell lines (SMMC-7721, MCF-7, HL-60, SW480, and A-549) and antibacterial activities against Bacillus subtilis, Escherichia coli, and Staphylococcus aureus.     

海洋细菌NTa发酵产琼胶酶条件的初步优化

利用选择性培养基从厦门红树林泥土样品中分离得到一株具有较高琼胶酶活力的海洋细菌NTa,通过16S rRNA分析,将该菌株归属为Stenotrophomonas sp.NTa.对该菌株发酵产琼胶酶的条件进行初步优化,结果表明:菌株NTa产琼胶酶的最佳碳源是琼脂,氮源是酵母浸膏,NaCl的质量分数为5%,在28 ℃发酵培养40 h,发酵液的酶活力达到1.98 U/mL,比优化前提高了37.35%.     

嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)R551-3四环素抗性消除的研究

嗜麦芽寡养单胞菌可共代谢降解烟碱类农药吡虫啉,并且对多种抗生素具有抗性.本文通过电穿孔法消除了该菌的四环素抗性,获得了四环素敏感型菌株R551-3Tcr-,可以用作遗传转化系统的受体菌,能被转化吸收遗传载体质粒pJB866H::ndhSL,构成一对很好的嗜麦芽寡养单胞菌遗传转化系统.同时HPLC结果表明所获得的四环素敏感型菌株羟基化吡虫啉的活性并未发生改变,为进一步开展敲除和回补共代谢途径关键酶的研究工作奠定了基础.     

土壤产几丁质酶菌株的筛选鉴定及产酶条件

利用几丁质为碳源,从土壤中筛选出3株产几丁质酶菌株,其中酶活最高的为一株革兰氏阴性菌.对该菌株应用16S rDNA法进行鉴定,结果为嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia).通过单因素优化法和均匀设计法实验,结果表明,以质量分数0.5%的胶体几丁质为碳源,质量分数1.0%的蛋白胨为氮源,及30℃和pH值为7.2,发酵60 h的条件是菌株的最合适产酶条件.此时,胞外几丁质酶酶活力最高.     

邻苯二甲酸酯高效降解菌的筛选及表征

以邻苯二甲酸二丁酯(DBP)为唯一碳源,筛选得到一株能够降解DBP的菌株.通过形态学观察、16SrDNA测序及系统发育分析,鉴定该菌株为寡养单胞菌(Stenotrophomonas sp.),命名为B3.首次报道了寡养单胞菌对邻苯二甲酸酯(PAEs)的降解作用.B3可高效降解0.05~50g·L~(-1)浓度范围的DBP,其中对0.05g·L~(-1) DBP的降解率为93.4%.经过优化后,在35℃、pH 8条件下对10g·L~(-1) DBP的降解率为95.8%.反应动力学研究表明,B3降解DBP符合一级降解动力学模型,对50g·L~(-1) DBP初始降解速率可达1.25g·L~(-1)·h-1.B3对其他4种常见的PAEs(邻苯二甲酸二(2─乙基己)酯、邻苯二甲酸二甲酯、邻苯二甲酸丁苄酯、邻苯二甲酸二乙酯)和苯胺、甲苯、邻苯二甲酸的降解率均在50%及以上.B3降解PAEs浓度范围宽、底物种类范围广,表明B3在PAEs的生物降解中具有良好的应用前景.     

嗜麦芽寡养单胞菌胞外蛋白酶的化学修饰

分别采用苯甲基磺酰氟(PMSF)、对-氯汞苯甲酸(PCMB)、N-乙咪唑(N-AI)、氯胺-T(Ch-T)、N-溴化琥珀亚胺(NBS)、2-巯基乙醇(2-ME) 6种化学修饰剂处理嗜麦芽寡养单胞菌胞外蛋白酶, 研究酶分子中氨基酸侧链基团与酶活性的关系. 结果表明Ch-T, NBS和2-ME能显著抑制酶活性, 而N-AI, PMSF和PCMB对酶活性的影响不大, 说明蛋氨酸残基、色氨酸残基和二硫键是酶活性的必需基团, 而酪氨酸残基、丝氨酸残基和巯基与酶活性无直接关系. 同时检测了EDTA和金属离子对酶活性的影响. 实验结果证实, EDTA, Mg^2＋, Ca^2＋, Hg^2＋和Cu^2＋能显著影响酶活性, 说明该酶为一种金属蛋白酶.     

嗜麦芽糖寡养单胞菌H002对铀的生物吸附

为处理铀元素造成的大规模水体污染,本研究拟采取生物治理的策略,以微生物吸附的方法来达到去除铀污染的目的.本实验从142株具有辐射抗性的细菌分离物中,筛选并鉴定到一株对重金属铀具有高度耐受能力的菌株H002;经16s rRNA和生理生化鉴定,确定H002属于嗜麦芽糖寡养单胞菌(Stenotrophomonas maltophilia).通过不同参数条件下的吸附实验表明,该菌在TGY培养基中培养12 h的细胞,对铀的吸附量最大;其中细胞表面的羧基参与铀的吸附;此外,不同阴、阳离子和表面活性剂会影响H002细胞对铀的吸附,其中十二烷基硫酸钠(Sodium dodecyl sulfate)可以促进对铀的吸附,而高浓度CO22-和HCO3-离子则抑制铀的吸附.本研究最终得到了一株具有铀耐受能力并能吸附铀的菌株H002,具有作为清除铀污染生物材料的潜力.     

猪源嗜麦芽窄食单胞菌的分离鉴定与耐药性研究

从临床上疑似嗜麦芽窄食单胞菌感染的病猪,进行病原分离、生化鉴定、特异性PCR鉴定和耐药性试验。结果获得了猪源嗜麦芽窄食单胞菌分离株20个．其中16株来源于病猪的呼吸系统;该菌对大多数抗生素具有耐药性且呈现多重耐药性．对复方新诺明最敏感。本研究从病原学角度证实猪嗜麦芽窄食单胞菌的临床感染并对临床用药有指导意义。     

Characterization and Antimicrobial Evaluation of Dilution-Stable Microemulsions Against Stenotrophomonas maltrophilia

The purpose of this study is to load glycerol monolaurate (GML) in a microemulsion system for the inactivation of Stenotrophomonas maltophilia. The influence of ethanol, propylene glycol (PG) and sodium benzoate (Na-Bz) on oil solubilization of GML microemulsions is clearly reflected in the phase behavior of these systems. One dilution-stable microemulsion formulation was determined and stable by the examination of the physical stability. Results showed that the microemulsion at 100 ppm, comparable to 3 mg/L ceftazidime, had much higher antimicrobial activities against S. maltophilia, with a major contribution of GML, and Na-Bz enhanced the antimicrobial activities as a hydrotrope.     

A New <Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis> Strain Producing Laccase. Use in Decolorization of Synthetics Dyes

Laccase activity was detected in a soil bacterium Stenotrophomonas maltophilia AAP56 identified by biochemical and molecular methods. It was produced in cells at the stationary growth phase in Luria Bertani (LB) medium added by 0.4 mM copper sulfate. The addition of CuSO₄ in culture medium improved production of laccase activity. However, one laccase enzyme was detected by native polyacrylamide gel electrophoresis. The enzyme showed syringaldazine (K _m = 53 μM), 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (K _m = 700 μM), and pyrocatechol (K _m = 25 μM) oxidase activity and was activated by addition of 0.1% (v/v) Triton-X-100 in the reaction mixture. Moreover, the laccase activity was increased 2.6-fold by the addition of 10 mM copper sulfate; the enzyme was totally inhibited by ethylenediaminetetraacetic acid (5 mM), suggesting that this laccase is a metal-dependant one. Decolorization activity of some synthetic dyes (methylene blue, methyl green, toluidine blue, Congo red, methyl orange, and pink) and the industrial effluent (SITEX Black) was achieved by the bacteria S. maltophilia AAP56 in the LB growth medium under shaking conditions.