Preparation of chitosan functionalized end‐capped Ag‐NPs and composited with Fe3O4‐NPs: Controlled release to pH‐responsive delivery of progesterone and antibacterial activity against pseudomonas aeruginosa (PAO‐1)

In this work, functionalized chitosan end‐capped Ag nanoparticles (NPs) and composited with Fe₃O₄‐NPs was prepared as pH‐responsive controlled release carrier for gastric‐specific drug delivery. The structure of prepared material was characterized by FE‐SEM, XRD, EDS and FT‐IR analysis. The loading behavior of the progesterone onto this novel material was studied in aqueous medium at 25°C and their release was followed spectrophotometrically at 37°C in seven different buffer solutions (pH 1.2, 2.2, 3.2, 4.2, 5.2, 6.2 and 7.2) to simulate intestine and gastric media which experimental results reveal more release rate in pH 1.2 (gastric medium) with respect to other buffers. This observation is attributed to dependency of the CS‐IMBDO‐Ag‐Fe₃O₄‐NPs and progesterone structure with buffer pH that candidate this new material as prospective pH‐sensitive carrier for gastric‐targeted drug delivery. On the other hand, the antibacterial properties of this material against gram‐negative bacterium pseudomonas aeruginosa (PAO‐1) in agar plates was studied and accordingly based on broth micro dilution the minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) with respect to standard CLSI in different concentrations of CS‐IMBDO‐Ag‐Fe₃O₄‐NPs was calculated. The results reveal that MIC and MBC values are 50 and 1250 μg/mL, respectively. In addition, extracts of Portulaca oleracea leaves was prepared and its antibacterial activity in single and binary system with CS‐IMBDO‐Ag‐Fe₃O₄‐NPs as synergies effect against PAO‐1 was tested and results shown that these materials have significant synergistic effect for each other.     

一株产吩嗪羧酸抗生素的甜菜内生细菌的分离与鉴定

从中国农业大学试验田中生长的健康甜菜肉质根内分离到1株分泌高活性抗真菌色素物质的优势内生细菌TCl,通过形态学、生理生化特征和16SrDNA系统发育分析,确认该菌株为绿针假单胞菌致金色假单胞菌亚种（Pseudomonas chlororaphis subsp．aureofaciens）。对TCI发酵分泌物经硅胶柱层析分离提纯后,采用薄层层析（TLC）和紫外一可见光谱扫描分析确认其主要活性成分为吩嗪一1一羧酸（Phenazine一1一Carboxylic Acid,PCA）。从TCl菌悬液浸种处理过的甜菜苗根内可以重复分离到完全相同的菌株,表明TCl的确可以在甜菜根内定殖,该菌株有可能被制成活菌制剂作为生物农药直接应用于宿主植物病害的生物防治。     

F3重组毒素的克隆表达及纯化

以pEKH-F3质粒为模板,PCR扩增F3片段,将F3插入克隆质粒PQE80L,转化大肠杆菌,筛选,获得含有F3的PQE80L重组载体的克隆.提取绿脓杆菌菌株染色体DNA为模板,PCR扩增绿脓杆菌外毒素A催化区,PE40.然后将PQE80-F3和PE40重组,得到表达PQE80L-F3-PE40载体.转化至大肠杆菌DH5a,BL21,表达融合蛋白F3-PE40.结果显示,大肠杆菌表达了融合蛋白F3-PE40,表达的融合蛋白量大概占菌体总体蛋白量的20%.通过质粒提取、PCR扩增构建F3-PE40表达载体转化大肠杆菌,成功地表达了融合蛋白F3-PE40,为大规模表达、纯化F3-PE40的进一步功能奠定了基础.     

培养基成分对海洋假单胞菌7-11产蛋白酶的影响

对海洋假单胞菌7-11(pseudomonas sp.7-11)产蛋白酶进行了培养基的优化,初步确定了促使蛋白酶产率提高的培养基的配方.经过26℃、140r/min、12 h培养,蛋白酶的相对产量可达201.7 IU/mL,比培养基优化前的26.8 IU/mL提高了646%.     

一种假单胞菌诱导CaCO3结晶

选用实验室自培育斯氏假单胞菌,通过测定pH值、电导率变化研究了细菌液、菌体液对碳酸钙结晶过程的影响,并通过SEM、XRD、红外等测试技术对生成的碳酸钙进行表征。 研究表明,斯氏假单胞菌细菌液与菌体液对碳酸钙结晶过程具有抑制作用,浓度增加,抑制作用越显著。 SEM、XRD和红外光谱的分析结果显示,细菌液可诱导亚稳态球霰石生成,菌体液能诱导出中孔方解石型碳酸钙。     

复合菌体及单一菌体催化的微生物燃料电池产电机理初步研究

利用淡水沉积物作为接种源构建了微生物燃料电池,考察苯酚对该微生物燃料电池性能的影响.结果表明,在淡水沉积物接种的微生物燃料电池中,电流的产生是由富集在电极表面的细菌引起的.苯酚降低了细菌消耗葡萄糖的速率,并在加入相同量葡萄糖的情况下,延长了产电时间.另一方面,实验还研究了一株从沉积物微生物燃料电池中分离出的单菌株的产电情况.该菌株在微生物燃料电池中需要借助自身代谢产生有电极反应活性的中间产物才能产电.GC-MS分析表明,中间产物中有吩嗪类物质,该类物质可在该细菌细胞与石墨电极之间充当电子传递介体,实现电子从细胞向电极的传递.     

耐碱性木聚糖酶发酵生产条件的研究

研究了不同培养条件下的产耐碱性木聚糖酶的情况 .类黄假单胞菌SQ153 产酶适宜条件为 :以4%的麸皮为碳源 ,以 0 .6 %的尿素为氮源 ,pH 10 .0 ,37℃ ,2 0 0r/ pm培养 14h ,其酶活力达 5 98IU/mL培养基 .在 5L自控发酵罐中 ,产酶达到同样水平 ,发酵周期缩短约 1h .     

Use of Eggshell Membrane as an Immobilization Platform in Microbial Sensing

Abstract

A new microbial biosensor was developed by immobilizing Pseudomonas fluorescens cells on eggshell membrane via physical adsorbtion. Bacteria-modified eggshell membrane was fixed tightly onto the surface of a carbon paste electrode (CPE) with a silicone rubber o-ring to construct a microbial biosensor. The measurements were based on the respiratory activity of the cells caused by oxygen consumption during bacterial metabolism. A mediated biosensor was also developed by ferrocene. As well as the response characteristics, stabilities and substrate specificities were investigated. Data were given as the comparison of mediated and unmediated systems.     

门多萨假单胞菌DS04-T对Poly(3HB-co-4HB)的降解研究

考察了门多萨假单胞菌DS04-T对Poly(3HB-co-4HB)的降解行为。以Poly(3HB-co-4HB)为唯一碳源,分别考察培养时间、培养温度、摇床转速、装液量、培养基起始pH值、接种量等因素对降解行为的影响。结合正交试验优化获得了菌株的最佳产酶条件:培养时间为28h,培养温度为30℃,培养基初始pH值为7.3,摇床转速为150r/min,培养基装液量为120mL(250mL三角瓶),接种量为1.5%(体积分数),此条件下菌株对Poly(3HB-co-4HB)的降解酶活力可达(26.2±0.7)U.mL-1。