培养基组成对米曲霉产氨基酰化酶的影响

氨基酰化酶是一类能够水解N-酰基-DL-氨基酸上酰胺键的酶。通过单因素实验和正交实验,对米曲霉产氨基酰化酶的培养基组成进行了优化,以提高其产量。确定较优培养基组成为:麸皮2%、尿素1.5%、磷酸二氢钠0.3%、硫酸镁0.35%、Tween-80 0.1%,在此培养基上得到的氨基酰化酶单位酶活可提高到440.51μmol.g-1.h-1。     

米曲霉氨基酰化酶拆分DL茶氨酸

DL-茶氨酸经乙酰化可生成N-乙酰-DL-茶氨酸，然后利用米曲霉氨基酰化酶拆分N-乙酰-DL-茶氨酸可以获得L-茶氨酸。本文对氨基酰化酶酶促反应也进行了初步研究，考察了反应温度、反应时间、pH值和底物浓度等因素对酶促反应的影响。实验表明，转化温度40℃、转化时间30h、pH为7．0和底物浓度0．2mol／L时，目的产物L-茶氨酸的产率最高。     

利用正交设计优化产氨基酰化酶的米曲霉的培养条件

采用正交实验设计L9（3^4）方法，利用摇瓶培养研究了产氨基酰化酶的米曲霉的培养温度、培养基pH和培养时间3个因素对菌体酶活的影响。结果表明，在培养温度29℃，培养基起始pH6．5以及培养时间50h。其酶活达595U／g以上。采用正交试验设计可以大大减小工作量，是一种高效率、快速、经济的实验设计方法。     

稀土和硫酸二乙酯联合诱变米曲霉产氨基酰化酶的研究

为获得高产氨基酰化酶的米曲霉菌株,选用稀土硝酸镧和硫酸二乙酯对米曲霉菌株CICC-2339进行联合诱变,经摇瓶发酵筛选到氨基酰化酶高产菌株SDESLa,其酶活为39.63 μmol·g-1·h-1,较出发菌株提高了39.50%,诱变效果显著.     

变色圈法筛选高产氨基酰化酶米曲霉菌株

将变色圈法用于筛选高产氨基酰化酶的米曲霉菌株,减少了筛选过程的工作量、缩短了筛选周期。通过实验验证了该方法在筛选高产氨基酰化酶米曲霉菌株时的可行性,并将其用于筛选经紫外线和硫酸二乙酯复合诱变的米曲霉。从突变株中获得了一株高产氨基酰化酶菌株W53,对其生长特性和产酶特性进行了研究。结果表明,米曲霉的氨基酰化酶是生长偶联型,菌体生物量和氨基酰化酶活力同时达到最高;突变株W53和出发菌株W19的产酶过程基本一致;W53拆分N-乙酰-DL-蛋氨酸的酶活力最高可达2569 U/g,比出发菌株W19提高了14.9%。     

固定化米曲霉氨基酰化酶反应动力学及其连续拆分DL-蛋氨酸实验

探讨了用明胶－戊二醛固定化米曲霉菌球氨基酰化酶反应动力学,对固定化菌体连续光学拆分DL－蛋氨酸进行了研究。当底物浓度小于200mmol/L时,没有底物抑制现象,此时拆分反应速度符合Michaelis-Menten方程。在37℃时,米氏常数和最大反应速率分别为11.9mmol/L和1.3mmol/L。在连续拆分反应中反应物体积流量为7.5ml/h,底物浓度为200和400mmol/L时,L－蛋氨酸的转化率分别为93％和78％。随体积流量的增加,L－蛋氨酸转化率降低。固定化菌体的操作半衰期为82d。     

固定化米曲霉氨基酰化酶反应动力学及其连续拆分DL–蛋氨酸实验

探讨了用明胶-戊二醛固定化米曲霉菌球氨基酰化酶反应动力学,对固定化菌体连续光学拆分DL-蛋氨酸进行了研究. 当底物浓度小于200 mmol/L时,没有底物抑制现象,此时拆分反应速率符合Michaelis-Menten方程. 在37℃时, 米氏常数和最大反应速率分别为11.9 mmol/L和1.3 mmol/L. 在连续拆分反应中反应物体积流量为7.5 ml/h,底物浓度为200和400 mmol/L时,L-蛋氨酸的转化率分别为93%和78%. 随体积流量的增加,L-蛋氨酸转化率降低. 固定化菌体的操作半衰期为82 d.     

Application of Box-Behnken design in optimisation for polysaccharides extraction from cultured mycelium of Cordyceps sinensis

A three-level Box-Behnken design, combined with the canonical and ridge analyses, was employed to optimise the process parameters for polysaccharide extraction from cultured mycelium of Cordyceps sinensis, one of the most valued traditional Chinese medicines and health foods. The critical factors selected for the investigation were extraction temperature, duration of time and number of times. The experimental results were fitted with a second-order polynomial equation by a multiple regression analysis and more than 96% of the variation could be predicted by the models. The canonical analysis of surface responses revealed that the three eigenvalues had different signs, indicating a saddle stationary surface. The optimal conditions for extraction of polysaccharides from the cultured mycelium of C. sinensis were determined, using the ridge analysis, as extracting 110 min at 88.9 °C for three times. Under the optimal conditions the corresponding response value predicted for polysaccharide production was 15.85%, which was confirmed by validation experiments.     

米曲霉菌体光学拆分N-乙酰-DL-苯甘氨酸的动力学研究

用液相培养基培养的米曲霉菌体直接拆分N 乙酰 DL 苯甘氨酸,并对该酶促反应进行了初步研究。考察了反应温度、pH值、金属离子、底物浓度、产物浓度等因素对反应的影响。结果表明,酶促反应的最适反应温度和pH值分别为52℃和7.0,当底物浓度小于200mmol·L-1时,反应符合Michaelis Menten方程,当底物浓度大于200mmol·L-1时,反应存在底物抑制现象。进一步研究发现水解反应也存在着产物抑制现象。     

米曲霉果糖基转移酶催化合成蔗糖-6-乙酸酯

对来自于米曲霉(Aspergillus oryzae)的果糖基转移酶催化蔗糖(s)与葡萄糖-6-乙酸酯(g-6-a)生成蔗糖-6-乙酸酯(s-6-a)的反应进行了研究。考察了底物质量比m(g-6-a)∶m(s)、反应温度、反应时间、pH、酶的质量浓度对反应的影响。单因素实验结果表明,最优的催化反应条件为:反应温度50℃、pH=6.2、m(g-6-a)∶m(s)=1∶3、反应时间48 h、酶的质量浓度为4.0 g/L。在该条件下,g-6-a的摩尔转化率可达26.69%。     

炭样小单孢菌抗生素对水稻白叶枯病菌的抑菌作用

[目的]研究炭样小单孢菌所产抗生素对水稻白叶枯病菌的抑菌作用.[方法]采用杯碟法和二倍稀释法探讨了炭样小单孢菌所产抗生素对水稻白叶枯病菌的抑菌活性.[结果]炭样小单孢菌所产的抗生素对水稻白叶枯病菌的抑菌直径达27.10 mm,最低抑菌体积分数与最低杀菌体积分数分别为0.78%和25%,在最低杀菌质量浓度下,抗生素对水稻白叶枯病菌作用18 h杀菌率为100%.[结论]炭样小单孢菌所产的抗生素对水稻白叶枯病菌具有较强抑菌作用.     

Production of Fumaric Acid from Raw Starchy Materials by Rhizopus oryzae with SSF

To reduce the manufacturing cost of fumaric acid production by fermentation, four kinds of raw starchy materials were investigated, cassava powder, corn powder, degermed corn powder and starch. By comparing the nitrogen contents in them, carbon/nitrogen ratio of raw materials was the key factor affecting the fumaric acid production by Rhizopus oryzae. Based on this point, degermed corn powder was selected as the best raw starchy material for fumaric acid production. In addition, the simultaneous saccharification and fermentation (SSF) process for fumaric acid production was optimized. With degermed corn powder as substrate at 100 g/L total sugar concentration, the fumaric acid production (35.51 g/L), yield (0.355 g/g) and productivity (0.493 [g/(L.h)]) were obtained via SSF process in a 7-L fermentor.     

Production of fructooligosaccharides by immobilized mycelium of Aspergillus japonicus

β-Fructofuranosidase (EC 3.2.1.26) in Aspergillus japonicus mycelium was immobilized by entrapment in calcium alginate gel. After immobilization, the enzyme was active over a wider pH range, and had improved thermostability. The total amount of fructooligosaccharides produced by immobilized enzyme was similar to that produced by a free enzyme system. A packed-bed reactor was employed for production of fructooligosaccharides at 42°C using the immobilized enzyme. The reactions were continued for 35 days and only 17% of enzyme activity was lost during this period.     

RP-HPLC测定红曲菌丝体中辅酶Q_(10)的研究

用反相高效液相色谱测定红曲菌丝体中辅酶Q10的含量。采用超声破碎细胞方法提取红曲菌丝体中辅酶Q10,色谱柱为Eclipse Pius C18(4.6*250 mm,5?m),流动相为甲醇∶无水乙醇=(V∶V)50∶50;流量为1 m L/min;进样体积为10μL;检测波长为275 nm;柱温为30℃。辅酶Q10在4.4453125~71.125?g/m L浓度范围内线性关系良好,回归方程为Y=8.7180X-4.7213,R2=0.9999,平均加标回收率为66.29%,RSD为2.41%。该法具有良好的线性关系,重现性好、精密度高、准确度好,可用于红曲菌丝体中辅酶Q10的含量测定。     

Lipolytic Enzyme Production by Immobilized Rhizopus oryzae

In this paper, a suitable technique as well as an adequate material for the immobilization of the fungus Rhizopus oryzae were investigated. This organism has been shown to have potential in the field of food aroma due to the production of extracellular lipolytic enzymes. However, an efficient production system at bioreactor scale for its application to the flavor compounds production is still needed. Among the supports studied, alginate beads were the best carrier materials, leading to the highest lipolytic activities of up to 400 U/L after 3 days of cultivation. Repeated batch cultures were carried out to improve cell concentration and lipolytic activity. The gel beads produced lipolytic enzyme under optimized conditions for consecutive batch fermentations without marked activity loss and deformation attained a maximum level of 715 U/L after three batches. The production of lipolytic enzyme by immobilized Rhizopus oryzae in a 2‐litre airlift bioreactor with the optimized conditions was evaluated. Lipolytic activities of 487 U/L were attained, operating in successive batches without operational problems and the bioparticles (the fungus grows in alginate beads) maintained their shape throughout fermentation.     

霉菌R.oryzae IFO催化大豆油脂合成生物柴油的研究

直接将霉菌R.oryzae IFO细胞应用于催化大豆油脂转酯合成生物柴油,并对其反应条件进行了优化.在反应所需甲醇分3次等量加入、醇油摩尔比11、反应温度35℃、缓冲液用量0.05mL/g油,缓冲液pH值范围3.6～6.98,生物柴油(脂肪酸甲酯)最终得率可达86%.