液态发酵条件下拟康宁木霉8985产纤维素酶能力初探

液态发酵条件下,以微晶纤维素为唯一碳源,比较了拟康宁木霉Trichoderma koningiopsis 8985和里氏木霉T. reesei QM9414产纤维素酶的能力。8985发酵12 h开始产生纤维素酶,36 h时酶活达到产酶峰值的50%,此时QM9414尚未诱导产酶。测定8985发酵84 h时上清液中滤纸纤维素酶、羧甲基纤维素酶、β-葡萄糖苷酶和木聚糖酶的酶活分别为1.06、3.62、1.80和6.67 IU/mL,分别是QM9414上述酶活的1.72、1.70、6.35和1.12倍。8985滤纸纤维素酶酶活的最适反应条件为pH 4.5,反应温度50 ℃,在Fe³⁺ (≤ 4 mmol/L)和Cu²⁺ (0-10 mmol/L)存在条件下酶活稳定。     

磁性Fe3O4微粒对葵花籽壳酶水解的影响研究

将磁化后的Fe3O4微粒添加于葵花籽壳酶水解过程中, 分析在不同的Fe3O4添加量和不同的添加方法下, 葵花籽壳酶水解过程中纤维素酶酶活﹑纤维素转化率及还原糖浓度的变化特征, 研究磁性Fe3O4微粒对纤维素酶水解葵花籽壳的影响。并通过考察酶水解反应前后水解液的表面张力值和pH值的变化, 探讨和分析磁性Fe3O4微粒作用下纤维素酶的磁效应机制。结果表明, 磁性Fe3O4添加量为0.5 g/L~2.0 g/L时, 对纤维素酶酶活的提高﹑还原糖浓度的增加和纤维素的转化在48 h后表现出较明显的促进作用。磁性Fe     

筛选两株稻杆降解放线菌

【目的】筛选能够同时降解纤维素、半纤维素、木质素的微生物菌株,并研究其对稻杆的降解效果。【方法】采用羧甲基纤维素钠、半纤维素平板水解圈法、苯胺蓝平板脱色法进行初筛;利用DNS法测定胞外酶活性;在含有2%稻秆的液体发酵培养基中摇瓶培养10天后,洗去菌体测定稻杆失重率、木质纤维素类物质降解率,同时测定稻杆断裂拉力进行复筛。【结果】筛选出两株能够同时高效降解木质纤维素的放线菌A3和A6,其纤维素和半纤维素酶活较高,最高纤维素全酶活、β-葡萄糖苷酶活、外切酶活和内切酶活分别为:12.84和12.85、6.23和6.53、24.56和17.80、14.00和18.80 U/mL;最高半纤维素酶活分别为:83.05和52.98 U/mL;木质素酶活较低,菌株A3和A6的木素过氧化物酶最大值为:12.72和14.67 U/mL,锰过氧化物酶最大值分别为:22.48和24.67 U/mL,漆酶最大值分别为:28.40和33.04 U/mL。经形态学、培养特征和分子生物学分析,两株菌株鉴定为链霉菌,对稻杆均有较好的降解效果,在第10 d后稻杆断裂拉力测定值分别比初始时降低62.67%和66.67%;稻杆失重率在31.50%和35.83%;菌株A3对纤维素、半纤维素、木质素降解率为38.73%、33.16%和20.68%,菌株A6为47.69%、28.64和22.59%。【结论】放线菌A3和A6对纤维素、半纤维素、木质素均具有降解作用,且酶活较高,具有较好的应用前景。     

工业纤维素诱导里氏木霉RUT C-30产葡聚糖酶的条件

为了研究工业纤维素诱导里氏木霉RUT C-30产葡聚糖酶的最佳条件,根据单因素实验结果,以工业纤维素、(NH4)2SO4和生物素为实验因素,滤纸酶活为响应值,进行中心组合设计,建立一个二次多项式数学模型,进行响应面优化,寻找最优产酶结果.经过优化,选出工业纤维素、(NH4)2SO4和生物素的添加量分别为39.485 g/L、6.232 g/L和249.872 μg/L,最高的滤纸比酶活为6.298 U/mL,实验验证,滤纸比酶活为6.118 U/mL,与预测值相差了2.86%.     

里氏木霉液体发酵产纤维素酶的研究

在摇瓶试验基础上,采用里氏木霉(Trichoderma reesei)HC-415菌株进行5L自控罐产纤维素酶深层发酵试验。在通气量为 0.2—0.6vvm、搅拌速度为 400r/min、发酵液pH控制在5.8—6.1的条件下,发酵液的羧甲基纤维素(CMC)酶酶活最高为325.0mg糖/ml,滤纸糖酶(FPA)酶活最高达17.9mg糖/ml。发酵周期为108h。所得冻干纤维素酶粉CMC酶活最高3111IU/g,FPA最高135IU/g ,对发酵液得率平均6.7g/L。酶活总收率CMC酶活平均78.2％,FPA酶活平均73.5％。     

长梗木霉纤维素酶的产生及提取

对长梗木霉(Trichoderma Longibrachiatum)ANU_3-958纤维素酶的产生及提取进行了研究。结果表明,固体曲培养144小时,固体曲与浸提液比例为1:7(W/V),采用硫酸铵分级沉淀法时所得纤维素酶活力最高。所得冻干纤维素酶粉经测定:羧甲基纤维素(CMC)酶活最高为2788.89IU/g,滤纸糖酶活(FPA)最高为79.44IU/g。相对固体曲得率平均为13.28%。     

紫孢侧耳栽培期基质中纤维素类的降解和有关酶活的变化

以棉籽壳为基质栽培紫孢侧耳（Ｐｌｅｕｒｏｔｕｓｓａｐｉｄｕｓ）,分别在菌丝长满期、菇蕾期、一茬菇和二茬菇时测定培养料中纤维素和半纤维素含量的变化以及纤维素酶（ＣＭＣ酶）和半纤维素酶活性的变化。结果表明：从接种到一茬菇时期,纤维素酶、半纤维素酶活不断升高,此后,迅速降低;基质中两种酶的底物含量降低速度以菇蕾一一茬菇时最大。菇蕾期后,两种底物的减少量多,菇蕾期以前较少。菇蕾期以前半纤维素降解稍多,菇蕾期以后纤维素降解较多。     

黑曲霉GD-6纤维素酶液体发酵条件的研究

采用黑曲霉 (Aspergillusniger)GD 6液体发酵生产纤维素酶 ,研究了碳源、氮源、培养基起始 pH值、接种量、摇床转速、通气量对该菌株产纤维素酶活力的影响。结果表明 ,GD 6的最适发酵温度为 2 8～ 3 0℃ ,产酶pH为 5 .5～ 6.0 ,摇床最适转速为 1 5 0r/min ,最佳接种量为 1 0 %。在以 6.0 %稻草粉为碳源、1 %豆饼粉为氮源时产酶活力最高。在最适培养条件下 ,发酵周期为 1 2 0h,发酵液中CMC酶活为 1 88.6U/mL ,FP酶活为 2 7.0U/mL。     

青霉的纤维素酶抗降解物阻遏突变株的选育

从土样中筛出一株生长快,产纤维素酶较高的斜卧青霉(Penicillium decumbens114-2)。能在全纤维素(hlocellulose)双层平板上形成清晰的透明圈。摇瓶培养的滤纸酶活可达8.8mg 葡萄糖/ml.h。 114-2菌株(其纤维素酶合成可为葡萄糖所阻遏)经UV和NG诱变处理后,在含葡萄糖的全纤维素平板上筛选到多株仍能形成明显透明圈的突变株。其中JN15和JU1在含葡萄糖的全纤维素液体培养基中,在残余葡萄糖浓度为 1％左右时,滤纸酶活可分别达到7.3 和13.9mg葡萄糖ml·h。这是出发株114-2和纤维素酶高产菌株木霉EA_3-867和QM9414所不能的。在不加阻遏剂的对比试验中,两个突变株的纤维素酶产量都比较高。其中,JU1的滤纸、CMC和棉花酶活分别达到33mg葡萄糖/ml·h,234mg葡萄糖/ml·h和86mg葡萄糖/ml·24h。     

pH值对绿色木霉(Trichoderma viride)产纤维素酶的影响

采用微晶纤维素为唯一诱导性碳源,对绿色木霉(Trichoderma viride)在摇瓶发酵过程中控制与不控制pH产纤维素酶进行比较.控制pH时胞外蛋白浓度为0.72 mg/mL比不控制pH时提高43%;FPA、EG、GB和CBH酶活为15.0U/mL,120.0U/mL,1.75U/mL,0.85U/mL分别是不控制pH时的2.1、2.3、11.7和1.7倍.在不同pH下测定纤维素酶液各酶活,表明pH值显著影响纤维素酶各单酶酶活.在pH2.7时,β-葡萄糖苷酶酶活仅为pH4.8时酶活的4%;pH回调试验结果表明β-葡萄糖苷酶对pH敏感,并在催化功能上发生不可逆变化.对纤维素酶液添加分离得到的各单酶,当添加β-葡萄糖苷酶时最多可以提高FPA酶活20%.因此β-葡萄糖苷酶是影响综合酶活的关键酶.通过拉曼光谱检测出β-葡萄糖苷酶在pH5.0有活性状态下,酶蛋白主链结构主要为a-螺旋和无规则卷曲;在pH2.0没有活性状态下,酶蛋白主链结构的无规则卷曲发生较大变化,a-螺旋也受到一定影响.这说明pH对β-葡萄糖苷酶构象的改变是造成其活性变化的主要原因.     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission

The molecular characterisation of species and genotypes of Cryptosporidium and Giardia is essential for accurately identifying organisms and assessing zoonotic transmission. Results of recent molecular epidemiological studies strongly suggest that zoonotic transmission plays an important role in cryptosporidiosis epidemiology. In such cases the most prevalent zoonotic species is Cryptosporidium parvum. Genotyping and subtyping data suggest that zoonotic transmission is not as prevalent in the epidemiology of giardiasis. Molecular characterisation of Cryptosporidium and Giardia is a relatively recent application that is evolving as new genes are found that increase the accuracy of identification while discovering a greater diversity of species and yet unnamed taxa within these two important genera. As molecular data accumulate, our understanding of the role of zoonotic transmission in epidemiology and clinical manifestations is becoming clearer.     

Synthesis and turnover of cerebrosides and phosphatidylserine of myelin and microsomal fractions of adult and developing rat brain.

The synthesis and turnover of cerebrosides and phospholipids was followed in microsomal and myelin fractions of developing and adult rat brains after an intracerebral injection of [U-14C]serine. The kinetics of incorporation of radioactivity into microsomal and myelin cerebrosides indicate the possibility of a precursor-product relationship between cerebrosides of these membranes. The specific radioactivity of myelin cerebrosides was corrected for the deposition of newly formed cerebrosides in myelin. Multiphasic curves were obtained for the decline in specific radioactivity of myelin and microsomal cerebrosides, suggesting different cerebroside pools in these membranes. The half-life of the fast turning-over pool of cerebrosides of myelin was 7 and 22 days for the developing and adult rat brain respectively. The half-life of the slowly turning-over pool of myelin cerebrosides was about 145 days for both groups of animals. The half-life of the rapidly turning-over microsomal cerebrosides was calculated to be 20 and 40 h for the developing and adult animals respectively. The half-life of the intermediate and slowly turning-over microsomal cerebrosides was 11 and 60 days respectively, for both groups of animals. The amount of incorporation of radioactivity into microsomal cerebrosides from L-serine was greatly decreased in the adult animals, and greater amounts of the precursor were directed towards the synthesis of phosphatidylserine. In the developing animals, considerable amounts of cerebrosides were synthesized from L-serine, besides phosphatidylserine. The time-course of incorporation indicated that a precursor-product relationship exists between microsomal and myelin phosphatidylserine. The half-life of microsomal phosphatidylserine was calculated to be about 8 h for the fast turning-over pool in both groups of animals.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Identification and composition of the tonsillar and anal enterococcal and streptococcal flora of dogs and cats

Enterococcus faecalis was the most frequently isolated enterococcal species from anal swabs and tonsils of dogs and cats, although in the anal samples from dogs Ent. hirae was found almost as often as Ent. faecalis. Most Ent.faecium strains from dog tonsils differed from those associated with humans and other animals in that they fermented sorbitol. Typical Ent. avium as well as atypical Ent. avium -like strains were seen in dogs, while the related species Ent. raffinosus was associated with cat tonsils. Enterococcus cecorum also occurred mainly in cats. Certain atypical strains, presumptively identified as Ent. cecorum , shared characteristics with Ent. columbae.
The most frequent streptococcal species in tonsils of cats and dogs were Streptococcus suis and Strep. canis. Streptococcus canis and Strep. bovis predominated in anal swabs. The canine Strep. suis differed from the common porcine strains in fermenting mannitol.
Forty-seven of the 288 isolates examined could not be identified or related to known species. The characteristics of two groups of these bacteria, provisionally called 'Ton 31 group' and 'O7 group' are described.