PVA降解菌的产酶条件及特性研究

目前对PVA生物降解研究重点逐渐转移到对PVA降解菌和PVA降解酶的研究开发上，随着对PVA降解高效新菌株的不断发现和PVA降解酶作用机理和分泌机制的深入了解，利用高效微生物或酶法治理PVA这类高聚物的污染将具有较大的应用潜力。本论文研究工作正是基于这种客观条件下进行的，对本实验室前期分离的PVA降解菌株P1、共生菌B1+B2、Pa、Pb为研究对象，重点研究了菌株P1和共生菌B1+B2的产酶条件和产酶特性，验证找出了影响菌株P1产酶的生长因子，论证了菌株B1+B2的产酶特性，优化得出了菌株B1+B2的最佳产酶条件；然后对共生菌B1+B2的PVA降解酶的稳定性进行了研究；最后研究了最佳组合菌的产酶特性和最佳产酶条件。主要研究结果如下： 1 通过对菌株P1产酶因子的研究，找出了核黄素是菌株P1产酶的必须因子，在以淀粉为碳源时，核黄素只是产酶的必须因子，而不是菌体生长的必须因子；在以PVA为碳源时，核黄素既是生长的必须因子，也是产酶的必须因子，是菌株P1的生长因子。 2 对共生菌B1+B2的产酶条件和产酶特性进行了研究，并通过正交实验找出了影响菌株产酶的主要条件和菌株产酶的最佳条件。 3 对共生菌PVA降解酶的稳定性进行了研究，确定了影响酶稳定性的主要理化条件。 4 通过对菌株降解性能的比较，确定菌株Pa、Pb、共生菌、P1的作为组合菌的组成菌，然后通过复配实验确定出菌株的最佳组合为菌株Pa、P1、共生菌，最后通过正交实验确定最佳组合菌的最佳配比。 5对影响组合菌产酶的因素进行了研究，通过正交实验确定了影响组合菌产酶的主要因素和最佳产酶条件。 本文通过对PVA降解菌株产酶条件和特性的研究，旨在为PVA降解菌生产酶制剂及进一步优化PVA降解菌在PVA废水治理中的应用提供理论和应用依据。 Now the PVA-degrading bacteria and polyvinyl alcohol-degrading enzyme are the key studies on the PVA biological degradation. It has great application potential using special bacteria and enzyme to treat pollution of PVA, with some high efficient Strain and enzyme were found. The study of this paper was based on that objective condition. The stain P1, symbiotic bacteria B1+B2, stain Pa and strain Pb were studied .The conditions of enzyme production and enzyme production characteristic of stain P1, symbiotic bacteria B1+B2 were our key study, we tested and verified the growth factor which effected enzyme production of strain P1, demonstrated enzyme production characteristic of symbiotic bacteria B1+B2, optimized and obtained the optimum conditions of enzyme production; then we studied the stability of polyvinyl alcohol-degrading enzyme of strain B1+B2; last the enzyme characteristic and the optimum conditions of alcohol-degrading enzyme production of optimum combination stains were studied. The main study results are below: 1. Through the study of enzyme production factor we found that lactoflavin is the necessary factor in strain P1 enzyme production. When we used starch to be carbon energy, lactoflavin is only the necessary factor of enzyme production, but not growth factor. When we used PVA to be carbon energy, lactoflavin was not only the necessary growth factor ,but also the necessary enzyme production factor.So it was the growth factor of strain P1 2. The enzyme production conditions and enzyme production characteristic of symbiotic bacteria B1+B2 were studied. Through the orthogonal experimental design, the main conditions which effected the enzyme production and the optimum conditions of enzyme production were obtained 3. Through the study of the stability of polyvinyl alcohol-degrading enzyme, the main physical and chemical conditions which effected the enzyme stability were 4. The stain P1,symbiotic bacteria B1+B2, stain Pa and strain Pb were selected to form combination bacteria. The stain P1,symbiotic bacteria B1+B2,stain Pa were the optimum combination through duplication experiment. Then the optimum ratio was obtained through orthogonal experiment. 5. Studied the factors which effected the polyvinyl alcohol-degrading enzyme activity, then through orthogonal experiment, the main factors and condition of enzyme production which effected the combination bacteria were achieved. The result of the study was valuable for the ferment of the PVA-degrading enzyme and the optimization of the PVA-degrading performance in the PVA wastewater.

三孢布拉氏霉发酵产β-胡萝卜素的研究

本论文研究了利用三孢布拉氏霉(Blakeslea trispora)发酵产β-胡萝卜素的培养条件。主要包括：发酵培养基的确定，发酵条件的优化。还考察了发酵菌丝体中β-胡萝卜素的提取方法及薄层层析等。 首先研究了培养基成分对三孢布拉氏霉发酵产β-胡萝卜素的影响。确立了玉米淀粉作为碳源，黄豆粉（热榨）作为氮源，棉籽油作为植物油的发酵培养基配方，其成分为：玉米淀粉 3％，黄豆粉（热榨） 2％，棉籽油 3％，KH2PO4 0.2%，MgSO4·7H2O 0.2%，维生素B1 0.002％，pH值6.0。 其次，通过比较不同的发酵影响因子，分别得到最适的条件：如三孢布拉氏霉正负菌接种比例为1.3：0.7，培养基pH值为7.0（灭菌后），发酵促进因子为Triton X-100。并采用正交试验法，确定其最佳发酵条件为正负菌接种比例1.3/0.7，发酵培养基pH为7.0，在培养基中添加表面活性基Triton X-100 0.08％。使该菌株产β-胡萝卜素的量达到0.73g/L，较初始发酵条件提高了3.3倍。 研究中还找到一个简便有效的对β-胡萝卜素的提取方法，选用盐酸-热处理法进行细胞破壁，并选用沸程为60～90℃的石油醚进行萃取。 用三孢布拉霉菌丝体内类胡萝卜索的石油醚提取液点样于硅胶G板，以丙酮：石油醚（5：95）为展开剂能将β-胡萝卜素与其它类胡萝卜索分离。该方法简便快速，并有一定实用价值。 The fermentative conditions of β-carotene by Blakeslea trispora have been investigated. These conditions include fermentation medium, the optimization of some fermentation factor. The extracting methods and the TLC of carotenoids were also researched. Firstly, the effects of composition of fermentation medium on the yield of β-carotene were studied. the results showed that the best fermentation medium was corn starch 3％，soybean power 2％，cottonseed oil 3％，KH2PO4 0.2%，MgSO4·7H2O 0.2%，vitamin B1 0.002％，pH value 6.0. Secondly, through compared some factors, such as different proportion of plus and minus strains, pH value, nonionic surfactants, respective best values have been obtained. The best proportion of plus and minus strains is 1.3:0.7, pH value of fermentation medium (sterilized) is 7.0, fermentation accelerant which acts as surfactants is Triton x-100. Farther on, the fermentative conditions were optimized through orthogonal experiment, the optimization showed that proportion of plus and minus strains is 1.3:0.7，pH value is 7.0, content of Triton x-100 is 0.08％. And the yield of β-carotene reached 0.73g/L, which was up to 3.3 times through the fermentation. In the extracting study, it has showed hydrochloric acid-heat treatment is a simple, convenient and effective extracting methods is which was used to destroy the cell wall, and the extracting organic solvent is petroleum ether whose boiling range is 60~90 ℃. In the TLC experiments, extracting contents in the petroleum ether were spotted in the silicagel plate, and the mixed liquor of acetone and petroleum ether (5：95) is developping agent, which can distinguish β-carotene from other carotenoids. It is a simple and quick technique.

HIRFL NB1控制系统的设计

通过对新B1聚束器的介绍 ,比较详细地分析了NB1腔体内部组件对谐振频率的影响 ,并以此为依据 ,阐述了NB1控制系统的设计应如何结合其物理特性 ,最终设计出了一套实时控制系统 .该系统包含RF发射机控制系统和拖动控制系统 ,均可手动和计算机对RF发射机和 5个步进电机实时控制

碳离子辐照细胞的相对生物学效率

研究了传能线密度 (LET)≥ 1 2 5.5keV μm的碳离子辐照小鼠黑色素瘤B1 6、人的宫颈癌HeLa、中国仓鼠肺V79、人的肝癌SMMC 772 1 4种细胞的相对生物学效率 (RBE) .得到了当LET =1 2 5.5keV μm时 ,RBE依赖于细胞种类并随细胞存活水平的升高而增加的关系 ,以及当LET≥ 1 2 5.5keV μm时 ,RBE随着LET的增大而变小的关系

DNA和细胞对重离子辐照诱导DNA双链断裂的敏感性比较

用倒转脉冲场凝胶电泳 (PIGE)比较研究了 50 Me V/u1 2 C6 +辐照小鼠 B1 6黑色素瘤细胞及其脱蛋白 DNA分子 ,从而诱导 DNA双链断裂 (DSB)的辐射敏感性。结果表明 ,不论辐照脱蛋白 DNA还是辐照完整细胞 ,诱导的 DNA片段释放百分比 (PR)都随着剂量的增加而增加 ,在超过 4 0 Gy后 ,PR趋向一个近似相同的准阈值～ 81 %。辐照脱蛋白 DNA诱导的 DSB产额为 0 .4 0 DSBs/1 0 0 Mbp/Gy,辐照完整细胞诱导的 DSB产额约为 0 .1 9DSBs/1 0 0 Mbp/Gy。说明脱蛋白 DNA分子比完整细胞对重离子辐照要敏感

阿佛曼链霉菌的~(12)C离子辐照效应及高产菌株选育；Research on Mutation Breeding of High-producing Strains from Streptomyces Avermitilis Irradiated by Ion Beam of ~(12)C~(6+)

选用12C6+离子辐照诱变阿维菌素B1a产生菌ZJAV-A1,研究其诱变效应。实验结果表明,12C6+离子辐照剂量50Gy时致死率97%,正突变率最高可达到34.2%。通过12C6+离子诱变处理,结合平板培养基及斜面培养基的正突变菌株筛选,最终获得一株稳定性良好,阿维菌素B1a组分产量稳定在4460—4588μg/ml之间,较出发菌株提高11.1%—14.7%的突变株ZJAV-Y1-203。

过柱纯化-高效液相色谱法测定玉米黄曲霉毒素

通过硅镁净化柱纯化,利用高效液相色谱法测定了玉米(Zeamays)中的黄曲霉毒素,在短时间(10min)内实现了黄曲霉毒素主要组分B1、G1、B2和G2的分离,确定了它们的回归方程和检测限。并在此基础上,结合目前研究现状,分析了该方法在应用中存在的问题,讨论了B1、G1、B2和G24种主要组分的出峰顺序。该试验结果将有助于高效液相色谱法在检测黄曲霉毒素应用中的日臻完善。

缓释肥料的研制及其缓效性评价

在模拟条件下通过筛选选择2种亲水性材料聚乙烯醇和聚丙烯酸钠,并用2种材料制造了6种包膜肥料SRF-A1、SRF-A2、SRF-A3、SRF-B1、SRF-B2、SRF-B3。并用水浸泡法和土柱淋溶法检验其缓效性,最后得出SRF-A3和SRF-B3在同种材料中具有较好的缓效性。

抗Her-2-scFv-SEC2融合免疫毒素的构建和功能研究(英文)

用基因工程方法,将金黄色葡萄球菌肠毒素 C2 与抗人表皮生长因子受体 HER-2 单链抗体 scFv-B1,以一连接短肽连接,构建融合免疫毒素 B-L-SEC2,并用改进的新型表达载体 pASK75-EX,在大肠杆菌 BL21(ED3)中表达. 以不溶性包涵体形式表达的目的蛋白经变性后以镍离子螯和层析纯化,并以透析法进行复性. 流式细胞术和 MTT 实验结果表明,纯化复性的融合免疫毒素 B-L-SEC2,在体外具有与 HER-2 过表达的靶细胞 SK-Br-3 特异性结合的活性,并对该细胞产生显著的特异性生长抑制作用.

天然吸附澄清剂在板栗壳棕色素精致中的应用

色泽是构成食品感官质量的一个重要因素,保持或赋予食品良好色泽是食品工业中亟待解决的重要问题。板栗壳棕色素是一种稳定性好,无毒副作用,甚至对人体具一定保健作用的天然色素,故可作优良的食品添加剂。然所含杂质则大大地降低其色价和品质,本文将板栗壳棕色素分别用101果汁澄清剂、ZTC1+1Ⅱ型澄清剂和壳聚糖进行絮凝处理,确定了各自的作用条件,并对其絮凝效果进行比较。101果汁澄清剂法:101果汁澄清剂1/125(v/v),温度室温(约15℃),色价7.27;ZTC1+1Ⅱ型澄清剂法:先B后A,B1/500(v/v),A1/1000(v/v),温度约40℃,色价9.39;壳聚糖法:1/125(v/v),温度约50℃,色价6.67。实验结果表明,三种方法对棕色素色价提高均有较好的效果,但总体上看,ZTC1+1Ⅱ型澄清剂法效果最好,色价高达9.39,故ZTC1+1Ⅱ型澄清剂法明显优于其它两种方法。

微生物混合培养及其应用

简述了混合培养微生物资源及其应用的研究进展。在长期的实验和生产实践中 ,人们发现很多生物过程是微生物纯培养不能完成或只能微弱进行的 ,必须依靠两种或两种以上的微生物共同培养完成。对于很多工业污染物、生物农药、纤维素、几丁质的生物降解 ,微生物混合培养是必要的 ;微生物混合培养可用于维生素C、维生素B1 2 、组氨酸、缬氨酸、L 苹果酸等发酵生产 ,还可用于药物的甾体转化、沼气发酵、湿法冶金等。混合培养的微生物资源应受到人们更多的重视。

不对称型双核半夹心结构铁化合物Cp′_2Fe_2(CO)_3Se_2C_2B_(10)H_(10)(Cp′=η~5-C_5H4Me)的合成、表征及结构

用碳硼烷的含硒锂盐 Li2 Se2 C2 B1 0 H1 0 (1 )与单茂羰基铁的氯化物 Cp′ Fe(CO) 2 Cl(2 )反应得到不对称型双核半夹心结构铁的化合物 Cp′2 Fe2 (CO) 3Se2 C2 B1 0 H1 0 (3 ) .X射线单晶结构分析表明其中一个铁原子是手性的 ,而且两个铁原子之间没有相互作用

人参四逆汤成分的研究

利用ESI/MS~n、LC/MS、MALDI-TOF/MS等技术,从人参四逆汤水煎液中分析和鉴定了10种二萜生物碱:苯甲酰中乌头碱benzoylmesaconitine、苯甲酰乌头碱henzoylaconitine、苯甲酰次乌头碱benzoylhypacoitine、尼奥灵neoline、附子灵fuziline、14-乙酰基塔拉地萨敏14-acetyl-talatisamine、苯甲酰次乌头碱亚油酸酯14-benzoylhypaconine-8-linoleate、苯甲酰去氧乌头碱亚油酸酯14-benzoydeoxyaconine-8-oleate、苯甲酰次乌头碱棕榈酸酯14-benzoylhypaconine-8-palmitate、塔拉地萨敏talatisamine和人参皂甙R_(a1)、R_(a2)、R_(b1)、R_(b2)、R_(b3)、R_c、R_d、R_e、R_(g1)、R_(g2)、R_(g3)、R_f等12种人参皂甙。脂肪酸酯型生物碱和苯甲酰类生物碱的提取组分有抗失血性休克作用和弱的正性肌力作用。人参皂甙的提取组分具有明显的负性肌力作用。

Metal ion and anion coordination in the thiamine-[Pt-II(NO2)(4)](2-) system. Structures of a metal complex, Pt(thiamine)(NO2)(3), and two salts, (H-thiamine)[Pt(NO2)(4)]center dot 2H(2)O and (thiamine monophosphate)(2)[Pt(NO2)(4)]center dot 2H(2)O

Reaction of thiamine or thiamine monophosphate (TMP) with K2Pt(NO2)(4) afforded a metal complex, Pt(thiamine)(NO2)(3) (1), and two salt-type compounds, (H-thiamine)[Pt(NO2)(4)]. 2H(2)O (2) and (TMP)(2)[Pt(NO2)(4)]. 2H(2)O (3), which were structurally characterized by X-ray diffraction. In 1, the square-planar Pt2+ ion is coordinated to the pyrimidine N(1'), a usual metal-binding site, and three NO2- groups. The thiamine molecule exists as a monovalent cation in 1 and a divalent cation in 2 while the TMP molecule is a monovalent cation in 3. In each compound, thiamine or TMP adopts the usual F conformation and forms two types of host-guest-like interactions with anions, which are of the bridging forms, C(2)-H . . . anion . . . pyrimidine-ring and N(4'1)-H(...)anion(...)thiazolium-ring. In 3, there is an additional anion-bridging interaction between the pyrimidine and thiazolium rings of TMP, being of the form C(6')-H . . . anion . . . thiazolium-ring. The salts 2 and 3 show similar hydrogen-bonded cyclic dimers of thiamine or TMP between which the anions are held. Results are compared with those of the other thiamine-platinum complexes. (C) 2001 Elsevier Science B.V. All rights reserved.

Host-guest-like thiamine-anion complexation in thiaminium bis(tetrafluoroborate)

In the title compound, 3-[(4-amino-2-methyl-5-pyrimidin-1-io)methyl]-5-(2-hydroxyethyl)-4-methylthiazolium(2+) bis(tetrafluoroborate), C12H18N4OS2+. 2BF(4)(-), the divalent thiamine cation (in the F conformation) is associated with BF4- anions via two characteristic bridging interactions between the thiazolium and pyrimidinium rings, i.e. C-H . . . BF4- . . . pyrimidinium and N-H . . . BF4- . . . thiazolium interactions. Thiamine molecules are linked by N-H . . .O hydrogen bonds to form a helical chain structure.