羰基还原酶基因工程菌产酶和不对称合成(R)-苯乙醇的条件优化

以羰基还原酶基因工程菌全细胞为催化剂,探讨不对称合成(R)-苯乙醇的各种影响因素,优化其产酶和转化条件。结果表明,优化后的发酵培养基为甘油10 g/L、蛋白胨10 g/L、酵母粉5 g/L,(NH_4)_2SO_4 5 g/L、NaCl 10 g/L,氨苄青霉素50μg/mL,pH 7.2;诱导剂IPTG的浓度为0.25 mmol/L,诱导时间为20 h,诱导温度为18℃,此条件下测得基因工程菌粗酶活最高为6.65 U/mL。建立的最佳转化条件:羰基还原酶基因工程菌发酵20 h后的细胞浓度为0.3 g/mL,转化体系初始pH为7.0,温度为37℃,辅助底物异丙醇浓度为10%,底物终浓度为60 mmol/L,转化时间为24 h。此时底物转化率最高可达98.88%,产物对映体过量值(e.e.值)为99.43%。反应体系扩增至3 000 mL后,产物e.e.值仍保持在99.0%左右,底物转化率可为92%以上,(R)-苯乙醇的产量也可达到6.67 g/L。     

米曲霉产淀粉酶固体发酵条件的优化

对本实验室筛选的米曲霉产淀粉酶固体发酵（SSF）条件进行而来优化。研究结果表明,所添加的6种碳源均抑制JLSP-14淀粉酶的合成;所补充的6种氮源中,酵母膏和硝酸铵可提高淀粉酶产量,以酵母膏为最佳,其最佳添加量为3．0％,其余4种氮源则抑制酶的生成：含水量以50％为最佳,发酵72h淀粉酶产量最高。     

提取果胶过程中双酶法去除马铃薯薯渣残余淀粉的研究

马铃薯渣中含有大量的淀粉,探索双酶法逐步水解马铃薯薯渣中的残留淀粉的研究,目的是解决马铃薯薯渣提取果胶的中性糖残留问题提高果胶纯度。结果表明,耐高温a-淀粉酶和高转化率糖化酶组合使用可有效的水解马铃薯薯渣中的残留淀粉,其最优工艺条件为:液化时耐高温a-淀粉酶的最适条件为,为pH为6.3,液化温度为95℃,加酶量为90u/g,液化时间为30min;高转化率糖化酶的最适条件为pH为4.3,液化温度为60℃,加酶量为198u/g,糖化时间为2.0h,在最适条件下,最终后续产品果胶的纯度上升至80%以上。     

淀粉酶产色链霉菌1628发酵培养基及发酵条件的优化

淀粉酶产色链霉菌1628(Streptomyces diastatochromogenes)是一株具有广阔应用前景的生防菌株,其合成的主要活性物质为核苷类抗生素——丰加霉素.为了提高淀粉酶产色链霉菌的发酵水平,本研究采用单因素和正交实验设计,对菌株1628的发酵培养基和发酵条件进行了优化,优化后的最佳培养基配方为:可溶性淀粉1.5%、黄豆粉5%、FeSO40.1%、CaCO30.5%、NH4NO3 0.3%、KH2PO4 0.3%,最适初始pH7.0.在供试的发酵条件内,装液量60mL/250mL,发酵温度28℃,摇床转速180r/min,发酵时间为92h.活性物质丰加霉素的产量最高156.17mg·L-1,比优化前提高了3.3倍.     

环氧基功能化块体SiO_2大孔材料对漆酶的固定化

采用后嫁接法在溶剂热条件下对新型块体SiO2大孔材料进行环氧基化改性,以环氧基功能化SiO2大孔材料作为载体,通过共价结合法固定化诺维信(Novozymes)工业级漆酶,对固定化条件进行了优化,研究了固定化酶与游离酶的酶学性质。结果表明,在固定化时间为4 h、pH值为4.5、初始酶液浓度为25 mg/mL时,固定化效果最好,固定化漆酶活力达到101.7 U/g;固定化漆酶的最适pH值为4.0,最适温度为50℃,其pH值稳定性和热稳定性都显著优于游离漆酶。固定化漆酶具有可重复操作的性质,与底物反应反复操作10批次后剩余活性为43.4%。     

响应面法优化改性芦苇生物炭对Se(Ⅳ)吸附条件研究

采用响应面法优化不同改性芦苇生物炭对水中硒的吸附条件,用Box-Behnken Design实验设计考察初始硒质量浓度、反应时间、吸附剂添加量和pH值等因素对硒去除率的影响。实验结果表明,各因素对芦苇生物炭(PB)去除硒的影响大小顺序依次为初始硒质量浓度、添加量、反应时间、初始溶液pH值;各因素对柠檬酸改性芦苇生物炭(CA-PB)和壳聚糖改性芦苇生物炭(CTS-PB)去除硒的影响大小顺序依次为初始硒质量浓度、反应时间、添加量、初始溶液pH值。PB吸附水中硒的最佳反应条件为初始硒质量浓度500μg/L,PB添加量为4.69g/L,初始溶液pH值为2.81,反应时间为2.50h,此时对硒的去除率达95.46%。CA-PB吸附水中硒的最佳反应条件为初始硒质量浓度479.76μg/L,CA-PB添加量为4.84g/L,初始溶液pH值为2.74,反应时间为2.59h,此时对硒的去除率达99.70%。CTS-PB吸附水中硒的最佳反应条件为初始硒质量浓度500μg/L,CTS-PB添加量为4.94g/L,初始溶液pH值为3.0,反应时间为2.50h,此时对硒的去除率达99.83%。     

北高灌蓝莓的离体快速繁殖研究

对北高灌蓝莓伯克利(Berkely)、蓝丰(Bluecrop)两个品种的离体快速繁殖进行了研究。试验表明:使用MS+Zt1.5mg/L+NAA0.2mg/L(MS:VB1增至0.4mg/L),培养基中添加蔗糖20g/L,琼脂粉5.6g/L,pH值5.2,接种后培养在温度为25±2℃,光强度为2500lx,光周期为16h进行增殖培养;采用改良1/2WPM基本培养基添加0.4mg/LIBA,蔗糖12g/L,琼脂粉3g/L,pH值5.0的生根培养基上培养为最优的北高灌蓝莓快速繁殖方案。     

黑曲霉内切聚半乳糖醛酸酶A基因在大肠杆菌中的表达及产物酶学性质研究

采用Trizol法提取黑曲霉(Aspergillus niger JL-15)的总RNA,RT-PCR扩增到黑曲霉内切聚半乳糖醛酸酶A基因(pgaA),pgaA全长1113bp,编码370个氨基酸残基.pgaA经EcoRI和NotI双酶切,定向插入到表达载体pET-32a(+)中,并转化Escherichia coli Rosetta-gami B(DE3),重组子经IPTG诱导,其表达产物(rePGA)果胶酶活性最高为637.0U/mg.酶学性质研究表明,rePGA的最适温度为60℃,热稳定性较差,最适pH为pH 5.0;Ca2+对其活性具有显著促进作用,而Fe3+和Mg2+对其活性具有显著抑制作用;SDS-PGAE结果表明,rePGA的相对分子质量约为40 500Da;rePGA的米氏常数(Km)和最大反应速度(Vmax)分别为4.05mg/mL和39.37μmol/(mL.min),rePGA能快速降低果胶溶液的粘度,符合其内切作用模式特点.     

软枣猕猴桃果胶的提取及其抑菌活性研究

目的为了优化软枣猕猴桃果胶的提取工艺,明确果胶及其酶解产物的抑菌活性。方法通过响应面法优化软枣猕猴桃果胶的提取工艺,通过滤纸片法测定果胶及其酶解产物对测试菌种的抑菌活性。结果得到了果胶的最优提取条件,料液比(g/mL)为1∶5,浸提pH值为2.3,浸提时间为80 min,浸提温度为70℃,在此条件下果胶提取率为(92.56±4.266)%;抑菌实验显示,果胶及其酶解产物对大肠杆菌的最低抑菌浓度分别为0.375 mg/mL和0.75 mg/mL,对金黄色葡萄球菌的最低抑菌浓度分别为3 mg/mL和0.75 mg/mL,对酵母菌的最低抑菌浓度分别为3 mg/mL和1.5 mg/mL,对青霉的最低抑菌浓度分别为0.75 mg/mL和0.375 mg/mL。结论酸提醇沉法可有效应用于果胶提取,软枣猕猴桃果胶及其酶解产物具有一定的抑菌活性,且酶解产物的抑菌效果优于果胶。     

超滤技术在碱性果胶酶浓缩工艺中的应用

运用从自然界中分离得到的一株具有产生特效高活性碱性果胶酶的软腐欧文氏菌株Erwinia Carotovora IFO 3830,通过发酵得到了含有高活性碱性果胶酶的发酵液由于果胶酶在该发酵液中浓度极低,仅为数μg/g左右.为了从大量的但含有目的酶浓度却极低的溶液中将目的酶分离出来,一般所使用的化工浓缩方法由于其操作温度较高使酶产品失活而无法运用.目前被普遍用于酶浓缩的硫酸铵沉淀法,不仅化学药品的消耗极大,而且后处理工艺又比较复杂;回收得到的沉淀酶制品中因混入了大量的培养基和其他杂质,导致后处理工序的更加复杂.本研究采用截留分子量为5 000和10 000(道尔顿)的超滤平面膜组件,可以直接从去除了菌体的发酵液中浓缩回收了目的产物碱性果胶酶,在浓缩率在20倍的条件下,取得了98.3%的高回收率.此外,还对超滤过程的表观阻力、浓缩率、渗透流束和浓缩液浓度等随着起滤过程的进行的变化规律进行了较为详细的研究     

Low cost removal of reactive dyes using wheat bran

In this study, the adsorption of Reactive Blue 19 (RB 19), Reactive Red 195 (RR 195) and Reactive Yellow 145 (RY 145) onto wheat bran, generated as a by-product material from flour factory, was studied with respect to initial pH, temperature, initial dye concentration, adsorbent concentration and adsorbent size. The adsorption of RB 19, RR 195 and RY 145 onto wheat bran increased with increasing temperature and initial dye concentration while the adsorbed RB 19, RR 195 and RY 145 amounts decreased with increasing initial pH and adsorbent concentration. The Langmuir and Freundlich isotherm models were applied to the experimental equilibrium data depending on temperature and the isotherm constants were determined by using linear regression analysis. The monolayer covarage capacities of wheat bran for RB 19, RR 195 and RY 145 dyes were obtained as 117.6, 119.1 and 196.1 mg/g at 60 degrees C, respectively. It was observed that the reactive dye adsorption capacity of wheat bran decreased in the order of RY 145>RB 19>RR 195. The pseudo-second order kinetic and Weber-Morris models were applied to the experimental data and it was found that both the surface adsorption as well as intraparticle diffusion contributed to the actual adsorption processes of RB 19, RR 195 and RY 145. Regression coefficients (R2) for the pseudo-second order kinetic model were higher than 0.99. Thermodynamic studies showed that the adsorption of RB 19, RR 195 and RY 145 dyes onto wheat bran was endothermic in nature.     

Utilization of organic by-products for the removal of organophosphorous pesticide from aqueous media

Sorption potential of rice (Oryza sativa) bran and rice husk for the removal of triazophos (TAP), an organophosphate pesticide, has been studied. The specific surface area were found to be 19+/-0.7 m(2)g(-1) and 11+/-0.8m(2)g(-1) for rice bran and rice husk, respectively. Rice bran exhibited higher removal efficiency (98+/-1.3%) than rice husk (94+/-1.2%) by employing triazophos solution concentration of 3 x 10(-5) M onto 0.2 g of each sorbent for 120 min agitation time at pH 6 and 303 K. The concentration range (3.2-32) x 10(-5) M was screened and sorption capacities of rice bran and rice husk for triazophos were computed by different sorption isotherms. The energy of sorption for rice bran and rice husk was assessed as 14+/-0.1 and 11+/-0.2 kJ mol(-1) and kinetics of the sorption is estimated to be 0.016+/-0.002 and 0.013+/-0.002 min(-1), respectively. Intraparticle diffusion rate was computed to be 4+/-0.8 and 4+/-0.9 nmol g(-1)min(-1/2). Thermodynamic constants DeltaH, DeltaS and DeltaG at varying temperatures (283-323 K) were also calculated.     

The adsorption of Cd(II) ions on sulphuric acid-treated wheat bran

The adsorption of Cd(II) ions which is one of the most important toxic metals by using sulphuric acid-treated wheat bran (STWB) was investigated. The effects of solution pH and temperature, contact time and initial Cd(II) concentration on the adsorption yield were studied. The equilibrium time for the adsorption process was determined as 4 h. The adsorbent used in this study gave the highest adsorption capacity at around pH 5.4. At this pH, adsorption capacity for an initial Cd(II) ions concentration of 100 mg/L was found to be 43.1 mg/g at 25 degrees C for contact time of 4 h. The equilibrium data were analysed using Langmuir and Freundlich isotherm models to calculate isotherm constants. The maximum adsorption capacity (qmax) which is a Langmuir constant decreased from 101.0 to 62.5 mg/g with increasing temperature from 25 to 70 degrees C. Langmuir isotherm data were evaluated to determine the thermodynamic parameters for the adsorption process. The enthalpy change (deltaH(o)) for the process was found to be exothermic. The free energy change (deltaG(o)) showed that the process was feasible. The kinetic results indicated that the adsorption process of Cd(II) ions by STWB followed first-order rate expression and adsorption rate constant was calculated as 0.0081 l/min at 25 degrees C. It was observed that the desorption yield of Cd(II) was highly pH dependent.     

Biosorption of nickel from protonated rice bran

In the present study biosorption technique, the accumulation of metals by biomass was used for the removal of nickel from aqueous medium. The rice bran in its acid treated (H(3)PO(4)) form was used as a low cost sorbent. The adsorption characteristics of nickel on protonated rice bran were evaluated as a function of pH, biosorbent size, biosorbent dosage, initial concentration of nickel and time. Within the tested pH range (pH 1-7), the protonated rice bran displayed more resistance to pH variation, retaining up to 102 mg/g of the nickel binding capacity at pH 6. Meanwhile, at lower pH values the uptake capacity decreased. The % age removal of nickel was maximum at 0.25 g of biosorbent dose and 0.25 mm biosorbent size. At the optimal conditions, metal ion uptake was increased as the initial metal ion concentration increased up to 100mg/L. Kinetic and isotherm experiments were carried out at the optimal pH 6.0 for nickel. The metal removal rate was rapid, with 57% of the total adsorption taking place within 15-30 min. The Freundlich and Langmuir models were used to describe the uptake of nickel on protonated rice bran. The Langmuir and Freundlich model parameters were evaluated. The equilibrium adsorption data was better fitted to Langmuir adsorption isotherm model. The adsorption followed pseudo second-order kinetic model. The thermodynamic assessment of the metal ion-rice bran biomass system indicated the feasibility and spontaneous nature of the process and DeltaG degrees values were evaluated as ranging from -22.82 to -24.04 kJ/mol for nickel sorption. The order of magnitude of the DeltaG degrees values indicated an ion-exchange physiochemical sorption process.     

Removal of Pb(II) ions from aqueous solutions by sulphuric acid-treated wheat bran

Sulphuric acid-treated wheat bran (STWB) was used as an adsorbent to remove Pb(II) ions from aqueous solution. It was observed that the adsorption yield of Pb(II) ions was found to be pH dependent. The equilibrium time for the process was determined as 2h. STWB gave the highest adsorption yield at around pH 6.0. At this pH, adsorption percentage for an initial Pb(II) ions concentration of 100mg/L was found to be 82.8 at 25 degrees C for contact time of 2h. The equilibrium data obtained at different temperatures fitted to the non-linear form of Langmuir, Freundlich and Redlich-Peterson and linear form of Langmuir and Freundlich models. Isotherm constants were calculated and compared for the models used. The maximum adsorption capacity (q(max)) which was obtained linear form of Langmuir model increased from 55.56 to 79.37mg/g with increasing temperature from 25 to 60 degrees C. Similar trend was observed for other isotherm constants related to the adsorption capacity. Linear form of Langmuir isotherm data was evaluated to determine the thermodynamic parameters for the process. Thermodynamic parameters show that adsorption process of Pb(II) ions is an endothermic and more effective process at high temperatures. The pseudo nth order kinetic model was successfully applied to the kinetic data and the order (n) of adsorption reaction was calculated at the range from 1.711 to 1.929. The values of k(ad) were found to be 5.82x10(-4) and 21.81x10(-4)(min(-1))(mg/g)(1-n) at 25 and 60 degrees C, respectively. Activation energy was determined as 29.65kJ/mol for the process. This suggest that the adsorption Pb(II) ions by STWB is chemically controlled.     

纳米Fe_3O_4/黑曲霉磁性微球对U(Ⅵ)的吸附性能及机制

在Fe_3O_4中加入黑曲霉培养基混合培养制备了Fe_3O_4/黑曲霉磁性微球,通过静态吸附试验,考察了pH值、温度、吸附剂用量、接触时间及U(Ⅵ)初始浓度等因素对Fe_3O_4/黑曲霉磁性微球吸附U(Ⅵ)效果的影响。结果表明:pH值是影响Fe_3O_4/黑曲霉磁性微球去除U(Ⅵ)的重要因素。pH=4、温度为25℃、U(Ⅵ)的初始浓度为10mg/L、Fe_3O_4/黑曲霉磁性微球投加量为1g/L时,Fe_3O_4/黑曲霉磁性微球对U(Ⅵ)的去除率达到98.89%,在吸附15h后趋于平衡。采用SEM、能谱分析、FTIR等手段分析了Fe_3O_4/黑曲霉磁性微球吸附U(Ⅵ)的机制。SEM-EDS结果表明,Fe_3O_4/黑曲霉磁性微球成功合成且呈网状结构;FTIR结果表明,Fe_3O_4/黑曲霉磁性微球与铀发生作用的主要基团有羟基、羧基、酰胺基等。     

磷石膏/聚氨基酸医用多孔复合材料的制备与表征

用化学共沉淀法合成了钙磷原子比(Ca/P)为1.50和1.67的磷石膏(PG), 用熔融聚合法制备四元氨基酸共聚物(PAA4), 用挤出发泡法制备了两种磷石膏/四元氨基酸共聚物(PG/PAA4)多孔复合材料。通过TGA、SEM、XRD、IR、EDS等对两种复合材料的组成结构进行了表征, 并研究了复合材料在磷酸缓冲液(PBS)中的体外降解性能。结果表明: 无机组分PG在两种复合材料中分布均一, 质量分数均为60%左右; PG/PAA4复合材料的孔径为100~400 μm, 孔隙率在60%左右; 多孔复合材料的有机和无机相之间有化学作用。PG/PAA4复合材料具有良好的体外降解性能, 其失重率随PG钙磷比的增加而增加, 降解液的pH值维持在6.9~7.4之间。PG/PAA4复合材料降解后, 其表面沉积了片状钙磷化合物, 推测该复合材料可能具有生物活性。     

响应面法优化CCMS / WG 复合包装膜制备工艺的研究

利用响应面分析法研究了CCMS/ WG 复合包装膜优化制备工艺,考察了谷朊粉、交联羧甲基玉米淀粉和丙三醇用量对CCMS/ WG 复合包装膜综合性能评定值的影响,得出了复合包装膜优化制备工艺的回归模型。结果表明,当谷朊粉用量为0. 65 g,交联羧甲基玉米淀粉用量为4. 77 g,丙三醇用量为1. 05 g 时,CCMS/ WG 复合包装膜性能最佳,其厚度为0. 096 mm,拉伸强度为20. 3 MPa,断裂伸长率为39. 1%,单位冲击破损能量为5. 73 J/ mm,水蒸气透过系数为3. 58 g·mm/ (m2 ·d·kPa),透氧系数为0. 69×10-15 cm3 ·cm / (m2 ·s·Pa),透光率为37. 6%,复合包装膜综合性能评定值最高达15. 73。     

炭/炭-陶瓷/炭多元复合材料的原位热压制备与性能

以纤维素和凹凸棒石(PG)为原料,在220℃下水热24h制备凹凸棒石/炭(PG/C)复合材料。采用浸渍-炭化工艺在炭/炭(C/C)复合材料中引入PG/C作为添加剂,一步热压对材料最终成型,原位获得C/C-陶瓷/C复合材料。研究了添加PG/C对C/C力学性能和抗氧化性能的影响。结果表明:PG/C在热压过程中转变为顽辉石/C,顽辉石/C通过"填充"和"桥联"起增强作用,顽辉石陶瓷表面负载纳米炭层有效避免了陶瓷相与基体炭间弱结合的产生。随着PG/C中表面负载纳米炭含量的减少,C/C的强度逐渐增加。当炭含量为13%的PG/C作为添加剂时,C/C的抗弯强度为263MPa,弹性模量为47GPa,相对于没有添加剂的C/C抗弯强度提高了45%,弹性模量提高了42%;相对于以PG作为添加剂的C/C抗弯强度提高了16%,弹性模量提高了27%。添加PG/C使C/C抗氧化性得到了提高;1 000℃下C/C的质量损失降低了12%~18%。     

In vitro binding capacities of three dietary fibers and their mixture for four toxic elements, cholesterol, and bile acid

Water-soluble dietary fibers from apple peels and water-insoluble dietary fibers from wheat bran and soybean-seed hull were used to evaluate their binding capacities for four toxic elements (Pb, Hg, Cd, and As), lard, cholesterol, and bile acids. The water-soluble dietary fibers showed a higher binding capacity for three toxic cations, cholesterol, and sodium cholate; and a lower binding capacity for lard, compared to the water-insoluble ones. A mixture of the dietary fibers from all samples - apple peels, wheat bran, and soybean-seed hull - in the ratio 2:4:4 (w/w) significantly increased the binding capacity of water-insoluble dietary fibers for the three toxic cations, cholesterol, and sodium cholate; moreover, the mixture could lower the concentrations of Pb(2+) and Cd(+) in the tested solutions to levels lower than those occurring in rice and vegetables grown in polluted soils. However, all the tested fibers showed a low binding capacity for the toxic anion, AsO(3)(3-).