响应面方法优化菊粉酶液体发酵培养基的研究

目的：为提高菊粉酶的产量。方法：运用Plackett-Burman设计法和响应面分析法,对Kluyveromyces S120液体发酵生产菊粉酶的培养基进行了优化。首先通过Plackett-Burman方法对8个相关影响因素的效应进行评价,并筛选出了有显著正效应的菊芋粉、玉米浆、(NH_4)H_2PO_4等三个因素,其他五个因素没有显著影响。然后根据Box-Behnken的中心组合设计实验和响应面分析方法确定了上述三个主要影响因素的最佳浓度。结果：在优化培养基下,菊粉酶产量为102．82u/mL,是优化前的2．1倍。     

产酸克雷伯氏杆菌发酵产2，3-丁二醇的培养基优化

采用不同设计方法相结合的策略对耐高糖产酸克雷伯氏杆菌（Klebsiella oxytoca）ME—UD-3-4发酵产2,3-丁二醇的培养基进行优化。首先在单因素实验的基础上采用Plackett—Burrnan设计法对影响ME—UD-3-4发酵产2,3-丁二醇的相关因素进行研究,筛选到3种有显著效应的因素（P〈0．05）：葡萄糖、玉米浆和MgSO4·7H2O。然后利用响应曲面法（Response Surface Methodology,RSM）对这3种因素的最佳水平范围进一步探讨;对得到的回归模型进行分析,得最佳条件（g／L）：葡萄糖220、玉米浆19和MgSO4·7H2O 0．4;在最佳条件下,发酵80h,2,3-丁二醇产量从原来的57．3 g／L提高到86．1 g／L,生产强度由0．72g／（L·h）提高到1．08g/（L·h）。     

黑曲霉纤维素酶高产固态培养基的快速优选研究

研究采用正交设计,对影响黑曲霉 Ｆ_２７产纤维素酶的 １０个营养因素在较少水平下进行初选,从中选出对产酶影响显著的４个因素．在此基础上采用均匀设计,在较多水平下进行较少次数的试验,将试验结果用微机逐步回归处理,得到最优回归方程,由回归方程计算出预测极值点和预测极值、优化出高产纤维素酶固态培养基配方．经试验验证,试验实测值和预测值基本相符, ＣＭＣａｓｅ活力平均达 ３７０５Ｕ／ｇ, ＦＰＡ活力平均达 ７．４Ｕ／ｇ．表明对于多因子试验,应用正交设计结合均匀设计的试验方法是快速、简便、可行的．     

响应面法优化Burkholderiasp．SYBCLIP—Y发酵产低温脂肪酶条件的研究

用响应面法对Burkholderiasp．SYBCLIP—Y液体发酵产低温脂肪酶的发酵条件进行了快速优化。首先利用Plackett—Burman设计对影响其产酶相关因素进行评估并筛选出具有显著效应的三个因素：牛肉膏,橄榄油,TritonX-100;用最陡爬坡路径逼近最大产酶区域后,利用响应面中心组合设计对显著因素进行优化,确定出牛肉膏,橄榄油,TritonX-100的最佳浓度分别为：牛肉膏31．8g／L、橄榄油21mL／L、TritonX-10036．55mL／L,优化后脂肪酶的酶活达到61．52U／mL,是优化前的2．62倍。     

公立医院医师满意度现状及对策探讨

目的：探讨公立医院医师满意度影响因素,为医院管理者了解医师的想法提供依据。方法：行设计员工满意度测评量表,开展预调查,对预调查量表采用可靠性分析及因子分析进行信度、效度检验,采用Likert五点测量法,对量表进行赋分,结果采用描述性分析及方差分析结果采用描述性分析及方差分析等方法。结果：不同职称医师满意度差异有统计学意义（P〈0．05）,不同医院医师总体满意度有差异（P〈0．05）;影响医生满意度的主要因素为工作回报、文化活动、工作本身．主观环境、科研氛围、工作本身．客观环境。结论：主治医师的满意度较低,医师总体满意度处于中等水平以上,提高医师薪酬,增加业余文化活动,提高医院职工就餐质量及环境．重视科研,开展多形式的培训是提高医师满意度的重要方法。     

高产蛹虫草菌株发酵培养基的研究

在单因素试验初步确定高产蛹虫草菌株发酵培养基的基础上,以蛹虫草茵丝体中腺苷含量为指标,进行11因素2水平Plackett—Burman试验设计试验,结合多元一次回归模型和F检验方法,筛选出发酵培养基中影响显著的组分酵母浸粉、蔗糖和维生素B1,采用旋转中心组合设计方法对这三个组分进行进一步优化,结合多元二次回归模型和响应面分析,获得高产蛹虫草菌株的最佳培养基（g／L）：蔗糖18．85、蛋白胨10、酵母浸粉18．97、KH2PO,3、MgSO4 3、维生素Bl0．235、ZnCl20．011、（NH4）2S0410。验证试验结果表明蛹虫草腺苷得率较单因素优化获得的发酵培养基提高了26．91％。     

响应面法优化低温淀粉酶发酵条件研究

目的：利用利用响应面法对低温淀粉酶发酵条件进行优化,确定最佳产酶条件。方法：使用DesignExpert软件的Plackett—Burman设计法对低温淀粉酶发酵条件进行了筛选,并利用响应面分析法进行了主要影响因素进行了回归分析。结果：筛出3个影响较大的因素,即蔗糖、蛋白胨、装添量;得到了各因素的最佳水平值,即蔗糖1．08％、蛋白胨2．02％、装添量87ml。结论：经3批发酵验证。其实验平均值38．1U／ml与预测值38．5U／ml非常接近,相关系数为98．8％。     

几种农艺因素影响小麦籽粒蛋白质含量的数学模型研究

用二次回归旋转组合设计研究了基本亩数（X_1）、基苗氮肥（X_2）、孕穗期氮肥（X_3)、过磷酸钙（X_4）和氯化钾（X_5）用量对小麦籽粒蛋白质含量的影响．结果表明,X_4、X_1和X_2对蛋白质含量有显著作用,因素之间存在着复杂的互作效应．利用计算机模拟选优,获得了本试验条件下蛋白质含量大于13％的5个农艺因素组合方案,X_1=172.35～186.75万／ha;x_2=116.6～123.6kg／ha;x_3=33.75～39.15kg／ha;x_4=333.9～448.8kg／ha;x_5=190.65～233.10kg／ha．     

响应面法优化玉米秸秆同步酶解发酵产乙醇条件

对汽爆玉米秸秆同步酶解发酵生产乙醇的条件进行优化。首先利用Fractional Factorial设计法对影响乙醇产量的7个因素进行评价,筛选出具有显著效应的3个因素,即反应温度、酶添加量、总反应时间,再以Box—Behnken设计法及响应面分析法确定主要因素的最佳水平,即反应温度37℃,每g纤维素添加纤维素酶32u,反应时间87h,此时乙醇体积分数达到3．69％。新工艺条件实验结果表明,乙醇体积分数在87h可达到3．76％,和原工艺相比,反应时间缩短了9h,乙醇体积分数提高了13％。     

响应面法优化波赛链霉菌JMC 06001发酵培养基

采用响应面法对产生抑菌活性物质的波赛链霉菌（Streptomyces peucetius）菌株JMC 06001的发酵培养基进行优化。首先采用Minnimum Run Equireplicated Res IV设计对初始发酵培养基的8个营养因素进行筛选,获得影响产生抑菌活性物质的3个主要影响因素：葡萄糖、大豆粉和NaCI;然后用最陡爬坡实验快速逼近最大响应区域;最后,结合Box-Behnken设计及响应面分析,确定主要影响因素的最佳浓度,得出该菌株产抑菌活性物质的最优发酵培养基配方为：葡萄糖1．2％,麦芽糖0．7％,蛋白胨0．9％,大豆粉1．4％,NaCl3．7％,CaCO3 0．1％,复合盐A液2．0％,复合盐B液0．1％,起始pH值7．0。用优化后的培养基发酵所得发酵液对敏感指示菌藤黄八叠球菌的抑菌圈直径达31．5mm,与预测值的相对偏差仅为1．59％,与用初始发酵培养基发酵所得发酵液的抑菌效果（抑菌圈直径26．5mm）相比提高了18．9％。     

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、...     

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.     

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.     

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.     

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。     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000