Screening of Penicillium species for occurrence of lectins and their characterization

Out of 15 Penicillium species screened for lectin activities, P. griseofulvum and P. thomii were found to possess mycelial lectin activity. None of the species displayed extracellular or cell surface‐bound lectin activity. Both species agglutinated rabbit erythrocytes. P. griseofulvum lectin showed specificity to human type O erythrocytes. While P. thomii lectin specifically agglutinated human type A erythrocytes. Highest lectin activities from P. thomii and P. griseofulvum were expressed after 8 and 7 days of growth, respectively. Lectins from both the species displayed a high binding affinity to chondroitin‐6‐sulphate, mucin, asialofetuin, D‐sucrose, and D‐trehalose. Ammonium sulphate at 50% saturation yielded 80% of the total lectin activity. Dialysis and ultrafiltration of the precipitates resulted in 1.79 and 3.46 fold purification of P. griseofulvum and P. thomii lectins, respectively. Both lectins showed pH optima between 7.0–8.0 and were stable near the neutral pH after 2 h. P. thomii lectin exhibited optimal activity at 35–40 °C, and P. griseofulvum lectin at 30–40 °C. P. thomii lectin showed a complete loss of activity above 40 °C, P. griseofulvum lectin was stable at or below 35 °C. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

青霉源抗菌肽类霉肽素的体外抑菌活性研究

目的研究抗菌肽类霉肽素（AF）的体外抑菌活性。方法通过抑菌圈法和二倍稀释法检测AF的抗菌谱和对金黄色葡萄球菌的MIC;通过检测在AF中传代菌的敏感性确定细菌是否容易对AF产生抗药菌;将AF和细菌在不同温度和pH环境作用后检测抑菌活性,明确AF发挥活性的最适温度和pH值。结果AF对大部分供试细菌和抗药菌有效,对金黄色葡萄球菌的MIC为0．8mg／ml,金黄色葡萄球菌在AF中传代200代后敏感性不变。在14℃到30℃之间AF的抑菌活性随温度升高而降低,在pH3到pH10之间AF的活性随pH值升高而降低。结论AF是一种对抗性菌有效的广谱抗菌肽,而且不容易诱导细菌产生抗药性。     

Morphology of Penicillium marneffei under oxidative stress in vitro

Penicillium marneffei is an intracellular pathogen; the mechanism allowing it to survive under oxidative stress remains unclear. For a better understanding of the response of P. marneffei to oxidative stress, the change in ultrastructure of this fungus before and after treatment with hydrogen peroxide was examined. A bamboo rat isolate and human isolate of P. marneffei were cultured on PDA at 25 °C and on BHI agar at 37 °C for 7 days respectively, with and without hydrogen peroxide; the morphology of strains was examined by optical microscopy and transmission electron microscopy. While comparing the human isolate with the bamboo rat isolate cultured without hydrogen peroxide, it showed no significant difference in ultrastructure. Microbodies were seen under transmission electron microscope in the yeast form, but could not be seen in mould form. After the strains were cultured with hydrogen peroxide, the mould form produced more rose red pigment; organelles of the fungal cells had been involved at different levels. Furthermore, the mould form of the human isolate with decreased conidia production and the yeast form with apoptosis could be observed. These results indicated that different strains of P. marneffei may have different levels of power to survive under oxidative stress.     

生物炭改善青霉素菌渣厌氧消化性能

目的 为提高青霉素菌渣厌氧消化性能。方法 在(35±2)℃,采用批式发酵工艺,在不同有机负荷菌渣中添加不同量生物炭进行厌氧消化。结果 添加生物炭,在有机负荷为2%VS和3%VS时,沼气产量比对照提高42.8%以上,甲烷含量比对照提前3~5d达到峰值,恢复pH值至7.0以上的时间明显缩短,VFA下降速度增快,氨氮浓度均在2500mg/L以下,VS去除率提高20%以上。有机负荷为4%VS时,沼气产量比对照提高了175.2%,但系统仍未正常运行。未添加生物炭,有机负荷为3% VS和4%VS时,发酵系统酸化,没有启动成功。结论     

产黄青霉Penicillium chrysogenum S-3-25人工海水培养基发酵次级代谢产物研究

目的 阐明南极深海来源真菌产黄青霉Penicillium chrysogenum S-3-25人工海水培养基的发酵次级代谢产物及其活性。方法 利用各种色谱技术分离纯化次级代谢产物,根据理化和波谱数据（核磁共振、质谱技术和Marfey分析）鉴定化合物结构,采用3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide（MTT）法评价细胞毒活性。结果 从真菌S-3-25人工海水培养基发酵产物中分离鉴定了8个单体化合物：2-[[(2S)-2-amino-1-oxopropyl] amino] benzoic acid (1),methyl 2-[[(2S)-2-amino-1-oxopropyl] amino] benzoate (2),2-[[(2S)-2-hydroxy-1-oxopropyl] amino] benzamide (3),3-(p-hydroxyphenyl)-N-methylpropionamide (4),4-hydroxycinnamamide (5),cyclo-(L-Pro-L-Tyr) (6),脑苷脂A (7) 及脑苷脂B (8)。细胞毒活性测试结果表明化合物1~8在50 μM作用浓度下对三种受试细胞（人乳腺癌MCF-7,人肺癌A549以及小鼠小胶质BV2细胞）的抑制率均低于50%。结论 从极地深海来源真菌产黄青霉S-3-25人工海水培养基发酵产物中分离得到8个单体化合物,其中化合物1和2为新天然产物,未见有文献数据的报道。化合物1~8均未呈现较强的细胞毒活性。     

经皮肤损伤感染马尔尼菲青霉菌致病力动物实验研究

目的研究马尔尼菲青霉菌经皮肤损伤途径感染小鼠致病力情况。方法将马尔尼菲青霉菌野生株和人感染株孢子悬液分别注入鼠尾皮内,观察接种后发病情况,于第15,50 d分批处死、解剖。结果马尔尼菲青霉菌导致小鼠皮肤感染发病率为100%,而85%小鼠皮损可自行消退痊愈,15%小鼠出现皮肤播散性感染;组织病理示:皮损处细胞性炎症反应在皮损中显著。从发病时间和早期病变严重程度比较,野生株致病力显著强于人感染株(P<0.05);但是从后期病变严重程度和自行痊愈率比较,野生株和人感染株致病力差异无显著性(P>0.05)。结论马尔尼菲青霉菌可以经皮肤损伤引起小鼠致病,其引起机体剧烈细胞免疫应答反应是致病力重要因素之一;野生株和人感染株感染早期致病力有差异,预后无差异。     

氟康唑诱导菌丝相马尔尼菲青霉菌产67.5kDa蛋白

目的探讨在体外加入氟康唑培养菌丝相马尔尼菲青霉菌的外分泌蛋白质变化情况。方法11株马尔尼菲青霉菌分别接种于含8μg/mL和不含氟康唑的30mL沙氏液基(SDB)中,25℃培养1周。E-test法测定不同培养液中菌丝对氟康唑的药敏。同时提取培养液中的蛋白质,聚丙烯酰胺凝胶电泳比较外分泌蛋白质差异。结果含氟康唑SDB中的马尔尼菲青霉菌菌丝悬液均变为玫瑰红色,不含氟康唑SDB中菌丝悬液颜色不变红。含氟康唑SDB中菌丝氟康唑MIC值显著增高(P<0.01),而且分泌67.5kDa蛋白质。结论在25℃沙氏液基中,8μg/mL氟康唑可以诱导马尔尼菲青霉菌产红色色素并分泌67.5kDa蛋白质。     

广泛播散型马尔尼菲青霉病患者的护理

报告了1例广泛播散型马尔尼菲青霉病患者的护理。主要内容包括:心理护理、皮肤护理、药物不良反应的观察、发热护理、营养护理、标本采集及配合诊断、控制感染、严密观察病情等。经过近3个月的精心治疗与护理,患者康复出院。     

Influence of oxidative stress and grains on sclerotial biomass and carotenoid yield of Penicillium sp. PT95

Oxidative stress and grains were evaluated for carotenoid production by solid‐state fermentation using Penicillium sp. PT95. When the fungus was grown at high oxidative stress, its sclerotial biomass and carotenoid content in sclerotia increased significantly with respect to low oxidative stress (P < 0.01). High oxidative stress also caused a statistically significant increase in carotenoid yield as compared with low oxidative stress (P < 0.01). Both the sclerotial biomass and the amount of carotenoid accumulated in sclerotia of strain PT95 were strongly dependent on the grain medium used. Among the grain media tested under high oxidative stress, buckwheat medium gave the highest content of carotenoid in sclerotia (828 μg/g dry sclerotia), millet medium gave respectively the highest sclerotial biomass (12.69 g/100 g grain) and carotenoid yield (10.152 mg/100 g grain). (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

具有免疫增强活性红树林真菌的分离及菌株WC1024的鉴定

目的 对采自海南省文昌市清澜港红树林的土壤样品进行真菌选择性分离,对其发酵液进行免疫增强活性筛选,并对活性较好的菌株进行鉴定.方法 利用四甲基偶氮唑盐(MTT)法检测菌株的免疫活性.通过菌落特征、显微形态、5.8SrDNA-ITS序列测定及其系统发育分析对菌株WC1024进行鉴定.结果 共分离得到真菌104株,活性筛选得到阳性菌株5株.其中菌株WC1024发酵液粗提物可刺激人外周血单个核细胞增殖,具有较强的免疫增强活性.通过各种指标鉴定菌株WC1024为疣孢青霉(Penicillium verruculosum).结论 添加青霉素和链霉素的海水马丁培养基用于海洋真菌的分类培养具有很好的选择性,首次鉴定发现1株具有显著免疫增强活性的疣孢青霉.