拟无枝菌酸菌M3-1降解丙酯草醚的特性及应用初探

丙酯草醚是我国自主研制的一种新型高效油菜田除草剂。采用现代分子生物技术和微生物学传统方法从长期施用嘧啶水杨酸类除草剂的农田中富集、筛选出以丙酯草醚为唯一碳源生长的高效降解菌株M3-1,并对其最适降解条件及在土壤中的应用进行研究。结果表明:(1)经形态、生理生化和16S rDNA鉴定,M3-1属于拟无枝菌酸菌(Amycolatopsis sp.)。(2)最适温度为35℃,pH为6.0,接种量为8%,在底物(ZJ0273)浓度为100～400 mg.L.1均具有一定的降解效果,且降解率随底物浓度的增加而降低,浓度高于300 mg.L.1时,对M3-1的降解效果有较强的抑制作用。(3)最适降解条件下,水溶液中培养25 d对浓度为100 mg.L.1丙酯草醚的降解率可达63.3%。(4)M3-1在土壤中对ZJ0273有很好的去除效果,处理25 d,其降解率可达53.7%。     

海洋栖热菌β-胡萝卜素15,15’双加氧酶原核表达

以海洋栖热菌（Oceanithermus sp.）DN-1基因组DNA为模板,通过PCR法扩增β-胡萝卜素15,15’双加氧酶BCMOTA基因,构建重组质粒,在大肠杆菌Escherichia coli BL21(DE3)中表达,并采用高效液相色谱（HPLC）法检测酶活性。结果表明,通过镍柱亲和层析和分子筛Sephacryl TM S-100,得到纯化重组β-胡萝卜素15,15’双加氧酶,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析结果表明,该酶分子质量约35 kDa;该酶的最适温度为50 ℃,最适pH值为8.0。在此优化反应条件下,生成569.3 mg/L全反式视黄醛,转化率达到89.5%。     

红曲霉菌种选育及固态发酵法生产Monacolin K研究

以紫红曲霉（Monascus purpureus）No．1为出发菌株，对该菌受制霉菌素影响的情况进行了研究，经紫外线诱变筛选得到1株具有稳定遗传性状的抗药性突变株AS．12，并对其产MonacolinK的发酵条件进行了摸索和研究，结果表明，在最佳的发酵条件下，物料厚度4cm，初始含水量50％，温度28℃，发酵12d，MonacolinK含量达2．186mg/g红曲干粉。     

降烟碱节杆菌发酵条件研究

在12种发酵培养基、不同温度、pH和培养时间条件下,对细菌生长能力和降烟碱能力进行测定,初步筛选出适宜4#菌株生长的为11号培养基,在5号、7号、9号培养基中4#菌株降烟碱能力强;适宜9#菌株生长的为8号培养基,在7号、9号、11号、12号培养基中9#菌株的降烟碱能力较强。适宜4#和9#的培养条件分别为30℃、pH6.5、48h和35℃、pH6.5、56h。     

链霉菌TD-1代谢产物对饲料中污染霉菌的抑制作用及稳定性

采用对峙培养法,检测分离于土壤中的链霉菌TD-1代谢产物对饲料中污染霉菌构巢曲霉（Aspergillus nidulans）、纯绿青霉（Penicillium verrucosum）、赭曲霉（Aspergillus ochraceus）、黄曲霉（Aspergillus flavus）、黑曲霉（Aspergillus niger）、米曲霉（Aspergillus oryzae）、茄腐镰刀菌（Fusarium solani）、桔青霉（Penicillum citrinum）、串珠镰刀菌（Fusarium moniliforme）的抑菌活性,结果表明：TD-1菌株代谢产物对以上霉菌具有较强的抑制作用,抑菌带宽度为10 mm左右.通过TD-1菌株的代谢过程曲线发现,该菌株发酵至第6 d抑菌率达90%左右.稳定性试验结果显示：该菌株所产生的活性物质具有较好的耐热性、耐酸碱性、抗紫外辐射能力.     

Identification and Characterization of a Novel Thermophilic,Organic Solvent Stable Lipase of <Emphasis Type="Italic">Bacillus</Emphasis> from a Hot Spring

A novel lipase gene lip256 was cloned and identified from the genomic library of hot spring strain Bacillus sp. HT19. The deduced amino acid sequence of lip256 has less than 32% identity to a predicted esterase (Cog1752) from Photobacterium leiognathi lrivu.4.1 and contains a novel motif (GTSAG) that differs from other clusters in the lipase superfamily. Following purification, a single band was obtained with a molecular mass of 33 kDa by SDS-PAGE, and the optimal temperature and pH for lipolytic activity of Lip25 were 70 °C and 9.0, respectively. Lip256 exhibited high activity at high temperatures, with 40% maximum activity at 80 °C and good stability at temperatures ranges between 50 and 80 °C. Additionally, the enzyme was highly stable in the presence of butyl-alcohol, glycerol, acetonitrile, pyridine, and urea. However, the presence of acetone, methanol, trichloromethane, petroleum ether, hexane, tert-butanol, isopropanol, dithiothreitol, ethylenediaminetetraacetic acid, polyhexamethylene biguanide, dimethyl sulfoxide, benzene, Triton X-100, Tween-20, Tween-80, and sodium dodecyl sulfate suppressed or absolutely inhibited enzyme activity. Furthermore, Ca²⁺, Mg²⁺, and Cu²⁺ suppressed enzyme activity, whereas Na⁺, Fe³⁺, K⁺, Fe²⁺, and Sr²⁺ enhanced enzyme activity. The unique characteristics of novel lipase Lip256, including its thermo-alkaliphilic performance, high tolerance toward metal ions, inhibitors, and detergents, and high stability in organic solvents, implied that this enzyme might be an interesting candidate for industrial processes.     

辅助能量物质柠檬酸促进ε-聚赖氨酸的合成

为提高ε-聚赖氨酸(ε-PL)合成能力,考察了柠檬酸对Streptomyces sp.M-Z18菌体生长和ε-PL合成的影响。研究发现,柠檬酸作为辅助能量物质,对ε-PL发酵过程影响显著。通过对柠檬酸添加时间、添加量和pH值的优化,确定了最佳柠檬酸添加方式,结合补料-分批发酵工艺,显著提高了ε-PL的产量。实验结果表明,在5L发酵罐中,维持pH为3.8,初始柠檬酸添加量为20 g/L时,分批发酵ε-PL的产量达到9.50 g/L,产率达到4.40 g/(L.d),较未添加柠檬酸发酵分别提高了46.4%和48.1%。采用添加柠檬酸的补料-分批发酵工艺,发酵168 h后ε-PL的产量达到22.89 g/L,是分批发酵的2.41倍。     

Comparative study on biosorption of Pb(Ⅱ) and Cr(Ⅵ) by Synechococcus sp.

The comparative study on adsorptions of Pb(Ⅱ) and Cr(Ⅵ) ions by free cells and immobilized cells of Synechococcus sp.was performed,in which different aspects including Zeta potential of the cells,the influence of pH,temperature and initial concentration of metal ions,as well as adsorption kinetics and mechanism were referred.The lyophilized free cells have a surface isoelectric point at pH 3,and the correlative experiment indicates that there is an electrostatic adsorption feature of Cr(Ⅵ) and Pb(Ⅱ).The immobilization of the free cells by Ca-alginate does not significantly modify the adsorption features of the biosorbent.The absorption processes of Cr(Ⅵ) and Pb(Ⅱ) on both free and immobilized cells are apparently affected by pH and the initial concentration of metal ions in the bulk solution,but are much weakly affected by temperature in the test range of 10-50 ℃.The slow course of biosorption follows the first order kinetic model,the adsorption of Pb(Ⅱ) obeys both Langmuir and Freundlich isotherm models,while the adsorption of Cr(Ⅵ) obeys only Freundlich model.FT-IR results indicate that carboxylic,alcoholic,amide and amino groups are responsible for the binding of the metal ions,and reduction of Cr(Ⅵ) to Cr(Ⅲ) takes place after Cr(Ⅵ) adsorbs electrostatically onto the surface of the biosorbents.     

天然抗氧化剂对裂壶藻油氧化稳定性影响的研究

采用Schaal烘箱法,以过氧化值和DHA含量变化为油脂氧化稳定性的评价指标,研究5种天然抗氧化剂对裂壶藻油氧化稳定性的影响。从中选择3种较优的天然抗氧化剂按一定比例复配得到复配天然抗氧化剂,并研究其对裂壶藻油氧化稳定性的影响。结果表明:5种抗氧化剂均能提高裂壶藻油的氧化稳定性,其中维生素E、迷迭香酸、茶多酚对裂壶藻油的氧化稳定性的提升较大;3种天然抗氧化剂复配配方为维生素E与迷迭香酸、茶多酚复配比例为2∶1∶2时对裂壶藻油的氧化稳定性的提升最大,在此条件下15 d后裂壶藻油的过氧化值为(30.25±0.57)mmol/kg,且该配比下DHA含量为(3.32±0.12)g/100 g,油脂的过氧化值低,DHA含量高,品质较好。     

塑料薄膜材料在微藻培养环境中的稳定性和生物附着行为

考察了6种薄膜材料在次氯酸钠(NaClO)溶液、HCl溶液(pH=2)和NaOH溶液(pH=12)中的稳定性及其在普通小球藻培养体系中的生物附着行为。结果表明,聚氯乙烯(PVC)在3种溶液中浸泡后420 nm处的透光率下降约50%,聚氨酯(PU)在NaClO溶液中、乙烯?醋酸乙烯酯(EVA)在NaOH溶液中浸泡后420 nm处的透光率下降10%?15%,聚乙烯(PE)、聚丙烯(PP)和聚对苯二甲酸乙二醇酯(PET)在3种溶液中浸泡后的透光率没有明显变化;6种薄膜材料在普通小球藻培养体系中均有微藻明显附着,浸泡7 d后透光率明显下降,其中PVC表面附着最严重,浸泡7 d后透光率接近0,附着物量随浸泡时间延长而增加,浸泡45 d后达3069 ?g/cm2;其它5种薄膜表面附着物量先增加后降低,PU和EVA在第15 d时、PE, PP和PET在第30 d时附着物量最大,分别为292, 375, 292, 194和236 ?g/cm2。