铅、锰对黄伞菌丝形态及细胞活性的影响

【目的】确定铅(Pb2+)、锰(Mn2+)对黄伞菌丝形态、结构及其菌丝活力的影响,并比较黄伞对Pb2+、Mn2+的适应性和耐受性。【方法】采用平板培养和液体培养方法,结合菌落形态和菌丝的电镜观察,测定菌落直径、菌丝鲜重,并以原子吸收分光光度计测定Pb2+、Mn2+含量。同时测定液体培养条件下菌丝的鲜重和胞外多糖产量以验证菌丝活力。【结果】不同Pb2+、Mn2+浓度下黄伞菌丝体形态变化明显,Pb2+、Mn2+的浓度≥500 mg/L可显著抑制黄伞菌丝的生长;高浓度Pb2+、Mn2+下(Pb2+≥700 mg/L、Mn2+≥2 000 mg/L)黄伞菌丝体锁状联合大量减少,且大小不一、分布不均,菌丝褶皱变形。黄伞在菌丝平板生长过程中,当Pb2+、Mn2+分别为100 mg/L时,黄伞菌丝生长速度最快。黄伞在液体培养过程中,当Mn2+浓度为300 mg/L、Pb2+浓度为50 mg/L时,菌丝鲜重以及产生的胞外多糖含量最大。【结论】Pb2+、Mn2+对黄伞的菌丝生长、菌丝活力及结构形态有较大的影响;黄伞对锰离子的适应性和耐性明显高于对铅离子的适应性和耐性。     

Alteration of cell-wall composition ofFusarium oxysporum by copper stress

A strain ofFusarium oxysporum tolerated copper in the growth medium at concentrations up to 600 mg/L. The optimum growth was obtained at 200 mg Cu/L. The mycelium acquired a blue color in the presence of copper. The copper content of isolated cell walls obtained from mycelium grown in the presence of 600 mg Cu/L was 1.5 times higher than that of cell walls obtained from mycelium grown at 200 mg Cu/L and it contained 2.2 and 3.3% copper at 200 and 600 mg Cu/L, respectively. The amount of protein and total sugars increased in both the mycelium and its isolated cell walls in the presence of copper in the growth medium, chitin was also increased in the cell wall, reaching its maximum amount at 200 mg Cu/L— about 2.4 times higher than without copper. Most of amino acid concentrations in the cell wall were increased in the presence of 200 mg Cu/L and decreased above this concentration. Isoleucine, leucine, tyrosine, phenylalanine, and arginine showed the highest increase at this concentration. The altered cell walls obtained from mycelium grown at 200 and 400 mg Cu/L could rebind individual metals more than the control cell walls could. Rebinding of individual metals was in the order Zn>Fe>Ni>Cu>Co. Rebinding of copper by isolated cell walls depended on pH and temperature.     

绿僵菌无纺布菌条的制作(I)液体种子培养条件

研究了金龟子绿僵菌菌株M a83液体培养最佳营养及环境条件,为无纺布培养提供良好的液体种子。以生物量为指标,通过液体摇瓶试验确定了绿僵菌M a83菌株液体种子培养参数,对不同的氮源、碳源及其浓度和初始pH进行单因素分析。结果表明,绿僵菌M a83菌株液体种子培养最佳营养条件为4%黄豆粉,2%白砂糖,0.5%MgSO4.7H2O,0.05g/LKC l,0.1g/L FeSO4.7H2O,1.0g/L KH2PO4,pH6.3~6.5。70L发酵罐培养,干生物量第三天可达35g/L,显著优于筛选前培养基(SDA,28g/L,a=0.05)。     

氮源对灵芝菌丝体液态深层发酵合成灵芝三萜的影响

以沪农灵芝1号为供试菌株,葡萄糖作为碳源,用硫酸铵、氯化铵、鱼粉蛋白胨、胰蛋白胨和酵母粉作为氮源,研究不同种类氮源对灵芝菌丝体液态深层发酵过程的影响。首先,确定了N-10酵母自溶粉作为发酵的氮源,降低了发酵的复杂性和不确定性;其次,考察N-10酵母自溶粉不同浓度对灵芝菌丝体发酵合成灵芝三萜过程中菌丝体的生物量、葡萄糖消耗、灵芝三萜产量等方面的影响,确定了N-10酵母自溶粉的适宜添加浓度。在此基础上,采用响应面中心组合设计,对4因素最佳水平范围进行研究,结果表明,葡萄糖、N-10酵母自溶粉、磷酸二氢钾和七水硫酸镁的含量分别为31.06g/L、2.76g/L、1.77g/L和1.99g/L时,灵芝三萜的理论产量为21.166g/kg干菌丝体,实际发酵产量提高到21.153g/kg干菌丝体。与原工艺相比,新工艺的灵芝三萜产量提高了6.22%。     

富铬蚁巢伞（鸡菌）液体培养(英文)

A species of mushroom, Termitomyces albuminosus, was cultured in liquid medium for production of chromium-enriched mycelium. The influence of chromium (Ⅲ ) on mycelial growth of T. albuminosus was investigated. An optimum medium composed of 5.6g/L yeast extract, 51.6g/L hydrolyzed rice, 2g/L KH2PO4, and 20mg/L chromium(Ⅲ ) with initial pH of 4.5 was obtained by using method of central composite design (CCD). After incubation of 84h, the maximal biomass of chromium-enriched mycelia reached 24.23g DMW(dried mycelial weight)/L with 272μg/g DMW chromium content in 500mL flasks containing 100mL medium with an inoculum of 8% on a shaker of 100r/min under an optimized cultivation condition at 28℃.     

古尼虫草小孢变种无性型原生质体制备及再生条件的研究

以从古尼虫草小孢变种Cordyceps gunnii var.minor上分离的无性型古尼拟青霉小孢变种Paecilomyces gunnii var.minor G106M为出发菌株,进行原生质体制备及再生条件的研究。将培养16h的菌丝体用溶壁酶和蜗牛酶的混合酶液于28℃酶解1.5~2h,原生质体产量可达3.74×107/mL。以培养12h的菌丝制备的原生质体在0.6mol/L氯化钠的SDAY培养基上再生率最高,为17.92%。     

Fermentation and microencapsulation of the nematophagous fungus <Emphasis Type="Italic">Hirsutella rhossiliensis</Emphasis> in a novel type of hollow beads

In this work, fermentation and formulation aspects of the nematophagous fungus Hirsutella rhossiliensis BBA were investigated. When incubated in 2% (w/w) glucose and 0.5% (w/w) yeast extract medium in a 1-L Erlenmeyer flask without baffles, heavy pellet formation was observed. Only 40% of the mycelium had a size less than 500 μm. When a flask with three baffles was used, the portion of mycelium <500 μm rose to 95%. In the next step, the influence of aeration rate and stirrer speed on production of finely dispersed mycelium in a stirred tank reactor was investigated. The best fermentation results were obtained at 0.4 vvm and 400 rpm stirrer speed with 90% mycelium <500 μm and 5 g/L biomass. Then, mycelium was microencapsulated in hollow beads based on sulfoethylcellulose (SEC). Experiments on the capsule nutrient reservoir showed that 15% (w/w) corn gluten and 0.5% (w/w) yeast extract could be replaced with 3% (w/w) autoclaved baker's yeast which was never used as capsule additive before. Radial growth of mycelium out of dried hollow beads containing 1% (w/w) biomass and 3% (w/w) baker's yeast was faster than for alginate beads containing equivalent amounts of biomass and yeast indicating a higher bio-control potential.     

忍冬桑黄和蛹虫草共发酵联产真菌多糖初步研究

忍冬桑黄和蛹虫草两种药用真菌均可产活性多糖,共发酵模式是产生新化合物或提高化合物含量或药效的潜在方式.本研究尝试用忍冬桑黄和蛹虫草共发酵联产真菌多糖,并对其共发酵所得的菌丝体多糖开展抗氧化活性试验.结果表明,当忍冬桑黄和蛹虫草预先分别发酵3d和1d后再同时接种共发酵,两菌可以较好地进行共生长,菌体总量和总多糖得率显著高...     

L-精氨酸高产菌红曲霉的筛选及发酵初步研究

对不同样品中的真菌进行了L-精氨酸高产菌株的筛选,从红曲米中分离出1株高产L-精氨酸菌株红曲霉H13,其初始发酵液中L-精氨酸产量为1.67 g/L.通过单因素实验对其液体发酵进行研究.结果表明,最佳碳源是可溶性淀粉,最佳浓度为10％,最佳氮源是蛋白胨,最佳浓度为3％,最佳pH值4.5,温度30℃,种子培养时间为18 h,摇瓶装液量50 mL/250 mL.转速为120 r/min,发酵时间7d.在此条件下进行发酵培养,所得发酵液产L-精氨酸4.08 g/L,菌丝体干重为10.32 g/L,菌丝体中L-精氨酸含量为24.17 mg/g.     

不同物质对白鬼笔菌丝生长的影响

本文研究了不同物质对白鬼笔菌丝生长的影响.采用平板培养法,以白鬼笔菌株Phallus impudicus-1为供试菌株,分析了 5种物质对白鬼笔菌丝生长的影响.供试碳源中,当以红糖为碳源时,白鬼笔菌丝生长速率最快,为1.445mm/d,蔗糖和葡萄糖次之;供试氮源中,当以酵母膏为氮源时,白鬼笔菌丝浓密,菌丝生长速率最快,...     

培养条件对头孢霉菌丝体脂肪酸组分的影响

研究了头孢霉(Cephalosporium sp.)菌丝体最大生产力和多不饱和脂肪酸形成积累的条件。菌丝体最适培养条件为：麦芽糖60g/L\,KNO33g/L、起始pH为60、500mL三角瓶装100mL培养基、接种25%、25℃培养10d则菌丝体达到最大干重。多不饱和脂肪酸形成积累的最适条件为：葡萄糖10～20g/L、(NH4)2SO4或NH4Cl 3g/L、培养基起始pH为40、500mL三角瓶装100mL培养基、接种10%～20%、10℃下照光培养。〖JP2〗因此,在整个生产流程中可采用不同条件分段掌握的技术原则。同时提出在多不饱和脂肪酸的形成和积累途径中油酸(18∶1)向亚油酸(18∶2)的转化是关键,为进一步探索最适培养条件和关键酶的调节提供依据。     

Effects of Cymbopogon nardus (L.) W. Watson essential oil on the growth and morphogenesis of Aspergillus niger

The growth inhibitory effect of Cymbopogon nardus (L.) W. Watson var. nurdus essential oil on Aspergillus niger (Van Tieghem) mycelium was determined on agar medium. The mycelium growth was completely inhibited at 800 mg/L. This concentration was found to be lethal under the test conditions. Essential oil at 400 mg/L caused growth inhibition of 80% after 4 days of incubation, and a delay in conidiation of 4 days compared with the control. Microscopic observations were carried out to determine the ultrastructural modifications of A. niger hyphae after treatment with C. nardus essential oil. The main change observed by transmission electron microscopy concerned the hyphal diameter and the hyphal wall, which appeared markedly thinner. These modifications in cytological structure might be caused by the interference of the essential oil with the enzymes responsible for wall synthesis which disturb normal growth. Moreover, the essential oil caused plasma membrane disruption and mitochondrial structure disorganization. The findings thus indicate the possibility of exploiting Cymbopogon nardus essential oil as an effective inhibitor of biodegrading and storage-contaminating fungi.     

Cadmium(II) Removal by a Hyperaccumulator Fungus <Emphasis Type="Italic">Phoma</Emphasis> sp. F2 Isolated from Blende Soil

A cadmium(II)-resistant fungus, strain F2, isolated from blende soil was identified as Phoma sp. by morphological study and internal transcribed spacer sequencing. This strain could accumulate 280 mg of Cd(II)/g dry weight mycelium. In liquid medium containing 163.8 mg Cd(II)/L, 96% of Cd(II) was removed by the actively growing mycelium. In addition, both oven-dried and lyophilized mycelium could effectively adsorb Cd(II). There were removed 91% and 46.2% of Cd(II) from 51.6 mg Cd(II)/L solution by lyophilized biomass and oven-dried biomass respectively. Transmission electron microscopy and energy-dispersive X-ray analysis showed the accumulation of Cd(II) in the mycelium cell walls. Our results demonstrated that Phoma sp. F2 was a hyperaccumulator for the removal of Cd(II) from contaminated soil and water.     

用灵芝发酵人参水煎液的工艺优化及其产物的抗疲劳活性研究

以灵芝为发酵菌株,对其发酵所采用的基础培养基和人参水煎液培养基分别进行筛选,并对人参水煎液的灵芝升罐发酵工艺及其发酵产物的抗疲劳活性进行了研究,结果表明灵芝最适基础培养基为葡萄糖5%,玉米浆0.3%,豆饼粉1%,糖蜜0.6%,MgSO4 0.075%,KH2PO4 0.15%;最适人参水煎液培养基为红参水煎液,浓度为40g/L;灵芝在红参水煎液中升罐发酵的最佳条件为：发酵周期58–62h。在此优化条件下,菌丝体干重达到7.0233g/L,胞外粗多糖含量达到0.3167g/L。人参的灵芝发酵产物的高、低剂量能明显延长小鼠游泳耗竭时间,并且降低游泳休息后小鼠的血乳酸含量,其中低剂量与空白组有显著性差异（P<0.05）,高剂量与空白组有极显著性差异（P<0.01）。说明红参水煎液的灵芝发酵产物具有显著的抗疲劳作用。     

一株祼脚菇属菌株产抗菌活性物的基础液体培养碳、氮源等优化及其对柑橘青绿霉菌的作用

江西是柑橘类水果的主要产区之一,青绿霉菌是导致柑橘采后腐烂的主要病害,化学杀菌剂可产生病原抗药性和残留,一株具有拮抗此病菌的祼脚菇属菌株0612-9被发现。为了探索其产活性物质的代谢培养和活性作用,采用菌丝生长速率法和显微观察法测定了其抗菌活性,单因素试验、Plackett-Burman、最陡爬坡试验和Box-Behnken试验响应面设计等优化出液体培养基组成和培养条件。结果显示:液体培养基组成和培养条件为:马铃薯279.94g/L,葡萄糖30g/L,麸皮8.87g/L,MgSO₄·7H₂O 0.47g/L,KH₂PO₄ 2g/L,发酵周期10d,接种量5 ml/100 ml,转速180 r/min,摇床温度26℃,装液量100ml/250ml,发酵液抗病菌活性从42.26%提高到55.77%。对意大利青霉和指状青霉的EC₅₀分别为69.25g/L和56.70g/L,最小杀菌浓度250μg/ml低于阳性对照物抑霉唑的500μg/ml,并且对病原菌菌丝体有一定的毒害作用。该祼脚属菌株有潜在开发成柑橘采后保鲜剂的价值。     

New nonsynthetic medium for Rhizobium culture production from wastes

A nonsynthetic medium was formulated for placement of mannitol fully by saccharified pea husk (Pisum sativum L.) and water hyacinth (Eichhoornia crassipes) with Trichoderma viride QM 9414 and molasses. Yeast extract was Partially replaced by proteolysed pea husk, water hyacinth, and mycelium of T. viride QM 9414 by boiling 4 hr with 5% (v/v) HCl. The rhizobial growth was equal in both standard yeast extract mannitol (YEM) and formulated nonsynthetic media. However, barring Rhizobium phaseoli (urid) E-6, the rhizobial counts in thenon-synthetic medium were higher then the counts in YEM medium. In the fermentor, rhizobial growth was also almost equal to YEM medium. These results indicated that costly ingredients like mannitol and yeast extract can be replaced by hydrolysates of pea husk, water hyacinth, mycelium of T. viride, and molasses.     

Study of the carbohydrate content in the mycelia of an active strain producing oxytetracycline and in an inactive mutant]

The content of carbohydrates in the mycelium of the active strain and inactive mutant of the oxytetracycline-producing organism under conditions favourable (starch medium) and unfavourable (glucose medium) for the antibiotic biosynthesis was studied. The mycelium of both organisms was fractionated and carbohydrate distribution according to the mycelium fractions and carbohydrate content in every fraction were investigated. No significant differences were observed between the active strain and inactive mutant with respect to the characteristics studied. The carbon source in the medium had the dominating effect on the chemical composition of the mycelium. The mycelium of both strains grown on the starch medium contained much more carbohydrates than that grown on the glucose medium. The carbohydrates of the mycelium grown on the starch medium were mainly found in fraction III and must be represented by polysaccharides.     

Production ofBeauveria bassiana [Deuteromycotina: Hyphomycetes] in different liquid media and subsequent conidiation of dry mycelium

Mycelium ofBeauveria bassiana can be grown in liquid culture, filtered, and the mycelium dried. After rehydration the mycelium sporulates. Two carbohydrate sources (sucrose and maltose), and one nitrogen/vitamin source (yeast extract) were tested for mycelium growth and subsequent conidial production. Maximum mycelium growth (12.31 mg/ml), in liquid culture, was in the sucrose (3.5%)/yeast extract (3.5%) medium, but mycelium from a maltose (2%)/yeast extract (0.75%) medium produced the maximum of 4.62×10⁶ conidia/mg dry mycelium after incubation in moist Petri dishes. Using the data on mycelium yield (in liquid culture) and conidial production (by dry mycelium) it is calculated that the sucrose (3.5%)/yeast extract (3.5%) and the maltose (2%)/yeast extract (0.75%) media produce most conidia per media volume (an equivalent of 3.52–3.72×10⁷ conidia/ml).      

16种茯苓菌株液体发酵产胞内多糖与三萜的比较研究

筛选茯苓高产胞内多糖和胞内三萜的优良液体发酵出发菌株。采用PDA富集固体平板培养与液体发酵培养测定菌丝体生长速率;采用液体发酵策略分析16种茯苓菌株产胞内多糖与胞内三萜的潜能。实验结果表明菌株生长于固体培养基与种子培养基的生长速率之间没有关联性;降低一级种子培养基初始pH值到4.0时能有效缓解茯苓菌株培养物褐化现象;AS5.137胞内多糖含量最高,达377.60±0.10 mg/g,而DB菌株显示出最高的胞内多糖产量,达1.01±0.13 g/L;Y1菌株胞内三萜含量最高,达83.89±4.28 mg/g,而Jingzhou28菌株胞内三萜产量最高,达136.63±26.66 mg/L。就生产茯苓胞内多糖与胞内三萜而言,AS5.137与DB菌株适合作为液体发酵产胞内多糖的出发菌株;Y1,Jingzhou28,Z(z)与Xingpinzhong菌株均较适合作为液体发酵产胞内三萜的出发菌株。     

产紫青霉中β-葡萄糖醛酸苷酶的诱导表达

针对产紫青霉(Penicillium purpurogenum)Li-3发酵生产β-葡萄糖醛酸苷酶存在的碳代谢阻遏现象,研究β-葡萄糖醛酸苷酶的高效诱导表达策略。在发酵条件优化的基础上,建立了新的产酶诱导工艺:葡萄糖的初始质量浓度5 g/L,在葡萄糖耗尽时加入20%诱导剂(10 g/L GL+1.2%Tween80)进行诱导,每24 h添加1次诱导剂,诱导72 h后立即转到40℃摇床发酵48 h。采用该工艺进行发酵,菌体出现了"二次生长"现象,比酶活从647.99 U/mL提高至2 356 U/mL,提高了近3倍。