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1.
Medium optimization for enhanced production of cytosine-substituted mildiomycin analogue （MIL-C） by Streptoverticillium rimofaciens ZJU 5119 总被引：1，自引：0，他引：1
Cytosine-substituted mildiomycin analogue (MIL-C) was produced effectively by supplementing cytosine into the culture of Streptoverticillium rimofaciens. In order to improve the yield of MIL-C, statistically-based experimental designs were applied to optimize the fermentation medium for S. rimofaciens ZJU 5119. Fifteen culture conditions were examined for their significances on MIL-C production using Plackett-Burrnan design. The Plackett-Burman design and one-variable-at-a-time design indicated that glucose and rice meal as the complex carbon sources, and peanut cake meal and NH4NO3 as the complex nitrogen sources were beneficial for MIL-C production in S. rimofaciens ZJU 5119. The results of further central composition design (CCD)showed that the optimal concentration of glucose, rice meal and peanut cake meal were 18.7 g/L, 64.8 g/L and 65.1 g/L,respectively. By using this optimal fermentation medium, the MIL-C concentration was increased up to 1336.5 mg/L, an approximate 3.8-fold improvement over the previous concentration (350.0 mg/L) with un-optimized medium. This work will be very helpful to the large-scale production of MIL-C in the future. 相似文献
2.
Studies on optimization of nitrogen sources for astaxanthin production by <Emphasis Type="Italic">Phaffia rhodozyma</Emphasis> 总被引：2，自引：0，他引：2
Ni H Chen QH Ruan H Yang YF Li LJ Wu GB Hu Y He GQ 《Journal of Zhejiang University. Science. B》2007,8(5):365-370
Fermentation ofPhaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using （NH4）2SO4, KNO3 and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L （NH4）2SO4, 0.49 g/L KNO3 and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively. 相似文献
3.
Bacillus subtilisis widely used for the productionof vitamins and other products including industrial en-zymes such as amylases, proteases and lipase. Butsyntheses of those products are repressed by the pres-ence of glucose, which is viewed as carbon catabolicrepression (CCR). InBacillus subtilis,CCR is main-ly mediated by the global regulator protein CcpAwhich was encoded byccpAgene[1]. It is therefore of crucial importance to relievingCCR ofBacillus subtilisfor industrial production. I… 相似文献
4.
本实验考察了基因工程茵Ecoli TOP10F’重组质粒pBAD／gⅢA—NTF2的稳定性。结果表明,该基因工程菌在连续传代30代后,质粒的目的基因片断没有缺失,具有结构稳定性;在无抗生素选择压力下,经传10代后,菌株的质粒没发生质粒丢失现象,20代后各代菌种质粒保有率低于100％,但50代后的菌种质粒保有率仍高达88％。因此,表现出一定的分离稳定性。该基因工程菌在优化培养基和优化条件下的整个发酵过程中,始终表现出良好的结构和分离稳定性。 相似文献
5.
Response surface methodology （RSM） was used to optimize the fermentation medium for enhancing pyruvic acid production by Torulopsis glabrata TP19. In the first step of optimization, with Plackett-Burman design, ammonium sulfate, glucose and nicotinic acid were found to be the important factors affecting pyruvic acid production significantly. In the second step, a 23 full factorial central composite design and RSM were applied to determine the optimal concentration of each significant variable. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the critical components were obtained as follows： ammonium sulfate 0.7498 （10.75 g/L）, glucose 0.9383 （109.38 g/L） and nicotinic acid 0.3633 （7.86 mg/L） with a predicted value of maximum pyruvic acid production of 42.2 g/L. Under the optimal conditions, the practical pyruvic acid production was 42.4 g/L. The determination coefficient （R^2） was 0.9483, which ensures adequate credibility of the model. By scaling up fermentation from flask to jar fermentor, we obtained promising results. 相似文献
6.
以饲料为原料筛选出一株既产乳酸又产芽孢的益生菌,经过分子生物学鉴定为凝结芽孢杆菌(Bacillus coagulans),命名为T-8。从培养基碳源、氮源、有机物浸出液、培养基pH、培养温度、培养时间等方面对菌株T-8展开发酵特性研究,结果表明,最优培养基配方为:玉米粉6 g/L、豆粕粉15 g/L、小麦麸皮浸出液60%、酵母粉5 g/L、蛋白胨10 g/L、氯化钠10 g/L,pH 5.0。以5%的接种量将种子液转接至最优培养基,在47℃的摇床中以200 r/min的转速振荡培养28 h,菌株T-8的细胞密度达到4.8×109 CFU/mL,芽孢率达到95.2%。本试验可为益生菌凝结芽孢杆菌应用于饲用微生物菌剂的工业化生产提供参考。 相似文献
7.
ZHU Yong GAN Jun-jiang ZHANG Guo-liang YAO Bin ZHU Wen-jie MENG Qin 《浙江大学学报(A卷英文版)》2007,8(9):1514-1520
In this work,rhamnolipid production was investigated using waste frying oil as the sole carbon source. By culture in shaking flasks,a naturally isolated strain synthesized rhamnolipid at concentration of 12.47 g/L and its mutant after treatment by UV light increased this productivity to 24.61 g/L. Fermentation was also conducted in a 50 L bioreactor and the productivity reached over 20 g/L. Hence,with a stable and high productive mutant strain,it could be feasible to reuse waste frying oil for rhamnolipid production on industrial scale. 相似文献
8.
将人细胞周期蛋白D1基因克隆入原核表达载体pET-20b中获得重组质粒pET-20b-eyeD,经酶切鉴定正确后转化大肠杆菌BL21PlaysS后获得表达菌株．该菌株经IPTG诱导后表达的目的蛋白有部分分泌到培养基上清中,将培养基上清中的蛋白沉淀后用Ni^2＋螯合柱进行纯化,最后可得到纯度达到95％以上的目的蛋白．蛋白电泳显示纯化蛋白的分子量约为33KD,Westemblot分析表明,在电泳胶的相应分子量处出现特异性条带,说明已经成功表达和纯化了重组人细胞周期蛋白D1． 相似文献
9.
通过PCR技术扩增出人铁蛋白基因,经酶切后与表达载体质粒pGEX-4T-2连接,重组质粒转化感受态大肠杆菌,利用菌落PCR、质粒双酶切、测序,证实成功地构建了人铁蛋白基因表达载体,利用IPTG对重组菌进行诱导表达,通过尿素洗涤纯化目的蛋白用于制备抗体． 相似文献
10.
生菜遗传转化体系的建立及口蹄疫抗原决定簇融合基因的导入 总被引：1，自引：0，他引：1
以散叶生菜大速生为试材,以MS为基本培养基,采用不同激素配比,确定生菜高效诱芽培养基为MS 1.5 mg/L 6-BA 0.2mg/L IAA;抗生素敏感性试验表明,筛选培养基中适宜的潮霉素选择压为20 mg/L,抑菌剂羧苄青霉素的适宜质量浓度为300 mg/L;通过根癌农杆菌介导的叶盘法将携带O型和A型口蹄疫抗原决定簇融合基因O21-O14-A21-HBcAg转入大速生散叶生菜,PCR检测转化率达50%。 相似文献
11.
对虾白斑综合症病毒（WSSV）编码与核酸合成代谢相关的核糖核苷酸还原酶（Ribonucleotide Reductases,RR）,与病毒DNA的复制有关．利用克隆技术将RR基因克隆到L4440载体,构建体内合成dsRNA的大肠杆菌HTn5工程菌．诱导该工程菌合成了RR基因的特异双链RNA（RR—dsRNA）和非特异双链RNA（gfP—dsRNA）,分别与WSSV混合共注射凡纳滨对虾,在感染72h后用病毒检测试剂盒提取DNA模板用于荧光定量PCR分析,结果显示RR—dsRNA能有效抑制WSSV病毒粒子的增值． 相似文献
12.
Xu YX Li YL Xu SC Liu Y Wang X Tang JW 《Journal of Zhejiang University. Science. B》2008,9(7):558-566
Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization.Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each sig-nificant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by mul-tiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x1 (urea)=0.163 (41.63 g/L), x2 (Na2CO3)=-1.68 (2.64 g/L), x3 (MGSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization. 相似文献
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14.
本研究对大肠杆菌进行紫外照射诱变后,获得了通过其内部的SOS修复系统产生的突变菌株,采用异丙醇沉淀法对各组菌种进行质粒DNA提取.通过测定各组OD值,经过比较与分析得出了经紫外处理10min后的大肠杆菌质粒DNA浓度最高,为215ug/ml,其次为正常组的190ug/ml.经紫外照射30min后的大肠杆菌质粒DNA浓度最低,仅为65ug/ml.紫外处理时间与质粒DNA浓度整体呈现下降趋势.初步推测是由于大肠杆菌SOS修复系统的RecA蛋白促进了不完全同源的DNA序列之间的重组,产生大量的错配碱基,从而引起突变,进而影响到其质粒DNA的浓度. 相似文献
15.
以拟南芥为材料,对热激蛋白HSP70基因片段进行了克隆研究,获得了最佳的分子克隆实验体系．首先提取拟南芥总DNA,接着在热激蛋白HSP70编码区设计引物,扩增出HSP70编码区的片断,再将HSP70编码区的片断加上具有EcoRⅠ酶切位点双链的人工接头,通过EcoRⅠ对其进行酶切．再与经过EcoRⅠ酶切并已经去磷酸化的PUC19载体通过T4连接酶进行连接,然后将连接产物置人到感受态大肠杆菌细胞中（即转化）,并让这种细胞在含有Amp＋／IPTG／X—Gal的LB培养基上生长．挑取培养基上蓝白斑中的白斑,进行质粒DNA提取,通过酶切质粒DNA,从而初步判断目的片段已被克隆． 相似文献
16.
以袋鼠爪花基部侧芽为外植体,进行了组织培养与快速繁殖技术的研究.结果表明:最佳诱导培养基为1/2 MS+BA 1.0 mg.L-1+NAA 0.05 mg.L-1+Ad 1‰+卡拉胶7 g.L-1+蔗糖20 g.L-1;最佳增殖培养基为1/2 MS+0.5 mg.L-1 BA+0.05 mg.L-1 NAA+Ad 1‰+卡拉胶7 g.L-1+蔗糖20 g.L-1;最佳生根培养基为1/2 MS+0.5 mg.L-1NAA+Ad 5‰+卡拉胶7 g.L-1+蔗糖20 g.L-1.炼苗后,移入泥炭和沙比例为2:1的基质中,移栽成活率高达90%. 相似文献
17.
根据GenBank数据库中猪圆环病毒Ⅱ型的基因组序列设计引物,采用PCR技术从病料基因组DNA中扩增出PCVⅡ河南地方株ORF4基因,全长180 bp,编码59个氨基酸.将该基因克隆至载体pGEX-4T-3中形成pGEX-4T-3-ORF4表达载体.经PCR、酶切和测序鉴定后,转化表达菌株BL21（DE3）诱导表达.SDS-PAGE结果显示：ORF4能够在大肠杆菌中表达,产物的分子量约为32 kD,且以包涵体形式存在.Western Blot检测结果显示,纯化后的ORF4蛋白能够与鼠抗6＆#215;His标签单克隆抗体发生特异性反应,为进一步研究PCVⅡORF4蛋白的特性与功能奠定了基础. 相似文献
18.
Biodegradation of crude oil by Pseudomonas aeruginosa in the presence of rhamnolipids 总被引：3，自引：0，他引：3
INTRODUCTION Oily wastewater, especially from oil field, hasposed a great hazard for terrestrial and marine eco-systems. The traditional treatment of oily wastewater,such as containment and collection using floatingbooms, adsorption by natural or synthetic materials,etc., can not degrade the crude oil thoroughly (Ollis,1992). So far, biodegradation suggests an effectivemethod. During biodegradation, crude oil is used asan organic carbon source by a microbial process,resulting in the brea… 相似文献
19.
以产没食子酸植物内生真菌CP01菌株为出发菌株,对影响菌体的生长代谢的葡萄糖添加量、培养温度、pH值、摇瓶最佳装液量、代谢前体物(苯丙氨酸)的添加量等因素进行了研究。结果表明:CP01菌株,在菌体培养5d的时候,没食子酸的含量和产量达到最高;葡萄糖含量为20g/L、培养温度为28℃、摇瓶装液量为100mL时,没食子酸的含量和产量达到最高。此外,苯丙氨酸含量为150mg/L时,没食子酸的含量和产量达到最高。 相似文献
20.
安诺兰的试管繁殖研究 总被引：1，自引：0，他引：1
利用我国云南、海南名优野生兰花品种安诺兰（ＡｎｏｔａｈａｉｎａｎｅｎｓｉｓＳｃｈｉｔｒ）的种子作外植体，采用试管无菌发芽的方法，成功地培养出试管苗．合适的种子萌发培养基为Ｈｙｐｏｎｅｘ３ｇ／Ｌ＋水解乳蛋白１ｇ／Ｌ＋活性碳０２％．较好的生根培养基为Ｈｙｐｏｎｅｘ３ｇ／Ｌ＋ＮＡＡ０５ｍｇ／Ｌ＋活性碳０２％．ＭＳ基本培养基不适合安诺兰的试管培养 相似文献