多杀菌素种子培养基及发酵培养基的优化

目的 以刺糖多孢菌CB11为试验菌株,以发酵培养基菌体浓度和多杀菌素产量为指标,通过单因素实验和正交试验对多杀菌素发酵的种子培养基的碳氮源成分进行优化,在此基础上通过响应面分析试验优化发酵培养基,找出最显著因素并确定其最佳值.方法 种子培养基碳氮源成分单因素实验和正交试验,发酵培养基响应面试验.结果 得到最优种子培养基配方为:葡萄糖10g/L,甘油5g/L,TSB25g/L,玉米浆10g/L,棉籽蛋白25g/L.在此培养基上生长的种子接入发酵培养基中所获得的多杀菌素产量达到411.26mg/L,较优化前水平(220.30mg/L)提高了86.68％.在此基础上,通过响应面分析试验优化了发酵培养基,得到对多杀菌素生产影响最显著的三个因素及最佳含最分别为葡萄糖66.6g/L、玉米浆14.6g/L、K2HPO4 2.5g/L,经验证获得的多杀菌素产量为544.60 g/L,较优化前产量提高了24.48％.结论 通过对多杀菌素发酵摇瓶种子培养基及发酵培养基的优化,找出了影响多杀菌素发酵的最显著因子及其最适含量,利用优化后的培养基发酵多杀菌素产量提高了86.68％,取得了较好的效果.     

海葵来源真菌 Cochliobolus lunatus 产大环内酯化合物LL-Z1640-2 的发酵优化

目的 对海葵来源真菌 Cochliobolus lunatus (TA26-46) 进行发酵优化以提高该菌产大环内酯化合物 LL-Z1640-2 的产量。方法 运用单因素试验设计和正交试验设计的方法,对菌株产 LL-Z1640-2 的碳源、氮源、初始 pH 值、发酵时间等发酵条件进行了优化。结果 得到了该菌株产 LL-Z1640-2 的优化发酵条件,其中,复合型培养基中可溶性淀粉 30.0 g.L–1,麦芽糖 20.0 g.L–1,硝酸钠 1.0 g.L–1,蛋白胨 3.0 g.L–1;初始 pH 6.0,培养时间 84 h。结论 在优化发酵条件下,LL-Z1640-2 产量可达到 (119.8±1.6) mg.L–1,比优化前提高了 3.6 倍。     

海洋真菌Penicillium sp.产抗肿瘤天然产物brefeldin A的发酵优化研究

目的 对海洋来源真菌Penicillium sp.产抗肿瘤天然产物brefeldin A的发酵优化,以期获得高产率目标化合物。方法 运用单因素实验设计,重点考察了培养方式、发酵培养基组成 (如碳源、氮源、盐度等)以及培养条件 (温度、摇床转速)等因素。结果 最佳发酵条件为：马铃薯200 g/L,麦芽糖10 g/L,淀粉20 g/L,海盐15 g/L,28 ℃, 摇床转速160 r/min,培养14天。结论 在该发酵条件下,brefeldin A的实际产量达到40 mg/L,较优化前提高了近10倍。     

海洋放线菌Streptomyces parvulus OUCMDZ-2554产放线菌素D的发酵条件优化

目的 对海洋放线菌Streptomyces parvulus OUCMDZ-2554产放线菌素D的发酵条件进行优化,提高放线菌素D的产量。方法 对培养基的组成、种龄、接种量、温度、装液量、pH、盐度和发酵时间等条件的研究,通过单因素和正交试验,选择最优发酵条件;并通过波谱分析及其理化性质确定放线菌素D结构。结果 最佳培养基为：K2HPO4 1.5 g、MgSO4 0.5 g、酵母浸膏5 g、可溶性淀粉22.5 g、陈海水1000 mL;最佳发酵条件：装液量150/500 mL（v/v）、种龄4天、接种量5%、盐度3%、起始pH 7.5、发酵温度28℃、摇床（180转每分）发酵12天。结论 以最佳发酵条件发酵,优化后放线菌素D的产量为优化前的3.6倍,达到364 mg/L。     

纺锤链霉菌SD-07产抗真菌抗生素CA-SD07发酵条件的优化

本文是对纺锤链霉菌SD-07(Streptomycese netropsis)发酵五烯大环内酯类抗真菌抗生素CA-SD07条件的优化.首先通过发酵液抑菌活力筛选获得理想的碳、氮源;其次单因素实验优化发酵条件至合理范围;最后用数理统计方法对发酵条件进行综合优化.结果 是Plackett-Burman筛选出影响CA-SD07产量的显著因素,中心组合试验设计(CCD)确定出其最适值.最后最适发酵条件为:葡萄糖35g/L,硫酸铵0.6g/L,豆粕粉2.5g/L,磷酸二氢钾1.00g/L,硫酸镁0.06g/L,pH5.9.在该培养条件进行5L发酵罐发酵验证,CA-SD07发酵单位达444.857mg/L,比优化前提高了375.4%.试验结果提示纺锤链霉菌SD-07菌株发酵CA-SD07的特性是接种量影响不大,对氧的需求比较高.     

用均匀设计优化达托霉素发酵培养基

目的:优化达托霉素的发酵培养基,提高达托霉素的发酵单位.方法:通过单因素试验设计考察不同碳源、氮源对达托霉素发酵单位的影响,筛选出适用于达托霉素发酵的碳源和氮源;再采用均匀设计对培养基中碳源、氮源的配比进行优化.结果和结论:在备筛选的碳源中,最适用于达托霉素发酵的是糊精,最佳用量为20g/L;最优的氮源是黄豆粉和蛋白粉合用,用量分别为30 g/L和5 g/L.优化后的发酵培养基可提高达托霉素发酵单位30%以上.     

海洋来源放线菌Actinoalloteichus cyanogriseus WH1-2216-6 产浅蓝霉素A的发酵条件优化

目的 对海洋来源放线菌Actinoalloteichus cyanogriseus WH1-2216-6 产浅蓝霉素A的发酵条件进行优化,提高浅蓝霉素A的产量。方法 对培养基的组成、接种量、起始pH、盐度、发酵时间、前体浓度及大孔吸附树脂添加量等条件的研究,通过单因素和正交试验,选择最优发酵条件;结果 获得了浅蓝霉素A的最佳发酵条件,培养基组成(可溶性淀粉20 g,甘油20 g,蛋白胨20 g,CaCO₃2 g,2-吡啶甲酸400 mg,XAD-16大孔吸附树脂50 g,陈海水1 L)、装液量150/500 mL(体积分数)、5 d种龄、3.3%盐度、起始pH=7.5、摇床(180 r/min,28℃)发酵12 d。结论 以最佳发酵条件发酵,优化后的浅蓝霉素A的产量为优化前的7.0倍,达到610.5 mg/L。     

响应面法优化链霉菌HY6-S36产星孢菌素的发酵条件

摘要：目的 优化链霉菌HY6-S36产星孢菌素的发酵条件,以提高星孢菌素的产量。方法 在单因素试验的基础上通过 Plackett-Burman试验、最陡爬坡试验、Box-benhnken(BB)响应面法优化发酵条件。结果 优化后,最佳培养条件为麦芽糊精 69.8g/L,豆粉49.6g/L,烟酸0.5g/L,碳酸钙4.0g/L,转速200r/min,pH7.2,装液量30mL/250mL,接种量5%,温度28℃,发酵 时间7d,摇瓶产量为996mg/L。经50L发酵罐放大培养,发酵效价达1063mg/L。结论 在此优化条件下,星孢菌素的摇瓶产量 提高了177%。     

酵母发酵生产谷胱甘肽的培养基优化

目的应用Plackett-Burman设计和球面对称设计实验,对酵母发酵生产谷胱甘肽的培养基进行优化。方法首先通过Plackett-Burman设计方法从10个因素中选择出对发酵产量影响较大的因素,即葡萄糖、酵母膏和半胱氨酸含量,然后用球面对称设计对这3个因素各取5个水平进行优化。结果最佳培养基组成为:葡萄糖23.64g/L,酵母膏29.07g/L,半胱氨酸1g/L,(NH4)2SO42g/L,蛋白胨5g/L,KH2PO41g/L,MgSO41g/L,NaCl2g/L。在优化条件下,发酵液中谷胱甘肽积累量可达162.3mg/L,比优化前产量提高约56.2%。结论证明用Plackett-Burman设计和球面对称设计寻求菌体积累谷胱甘肽的最佳培养基组分是可行的。     

南极放线菌N-1产抑菌成分的响应面优化

目的本研究旨在对南极放线菌N-1的发酵工艺进行优化,以提高发酵液中抑菌成分的活性。方法本实验以抑菌活性(琼脂扩散法)为指导,对该菌的碳源、氮源、无机盐等培养基组成进行单因素考察,以期得到显著性影响因素,然后利用Plackett-Burman试验设计,最陡爬坡设计以及Box-Behnken组合设计对显著性影响因素进一步进行优化。结果通过这些条件的优化,最终得到南极放线菌N-1的最佳发酵条件为小米12.68g/L、NaNO36.67g/L、MgSO4.7H2O 0.31g/L、FeSO4.7H2O 0.03g/L、黄豆粉7.50g/L,CaCO30.60g/L,NaCl 20g/L,pH=6.0,温度28℃,转速200r.min-1。与优化前相比,发酵液的活性提高了5.94倍;菌体湿重提高了1.55倍。结论利用响应面法(RSM)对南极放线菌N-1的发酵条件进行优化之后,提高了该菌的菌体密度的同时,更加提高了发酵液中抑菌活性物质的抑菌能力,为活性天然产物的发现奠定了较好的基础。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.