杜连祥
主要从事工业微生物和生物制药方面的研究。
个性化签名
- 姓名:杜连祥
- 目前身份:
- 担任导师情况:
- 学位:
-
学术头衔:
博士生导师
- 职称:-
-
学科领域:
微生物学
- 研究兴趣:主要从事工业微生物和生物制药方面的研究。
杜连祥,男,1941年8月出生,河北青龙人。1964年毕业于天津轻工业学院发酵工程专业。现任天津科技大学生物工程学院教授,博士生导师。国家优秀教师,天津市劳动模范。享受政府特殊津贴,被天津市政府授予“工业发酵工程”授衔专家。担任中国微生物学会理事,中国啤酒工业协会理事,中国微生物学会工业微生物委员会副主任等社会兼职。
杜连祥教授主要从事工业微生物和生物制药方面的研究,先后主持承担国家自然科学基金项目“油为碳源抗生素合成机理及发酵动力学研究”等国家、省部级科研项目16项。获1987年天津市科技进步三等奖,1989年科技进步二等奖;1991年科技进步三等奖;1992年科技进步一等奖;1990年河北省科技进步二等奖;2000年天津市科技进步二等奖;2004年天津市科技发明二等奖,北京市科技进步二等奖;2005年天津市科技进步二等奖。共授权发明专利3项。在《生物工程学报》、《食品与发酵工业》等核心期刊上发表高水平论文数十篇,并有多篇文章被SCI收录;主编《工业微生物学实验技术》等教材。
-
主页访问
1422
-
关注数
0
-
成果阅读
429
-
成果数
10
【期刊论文】Adaptive response of Saccharomyces cerevisiae to hyperosmotic and oxidative stress
杜连祥, Fuping Lu a, *, Yu Wang a, Dongqing Bai a, b, Lianxiang Du a
Process Biochemistry 40(2005)3614-3618,-0001,():
-1年11月30日
After being pretreated with a sublethal dose of either oxidative or hyperosmotic stress, Saccharomyces cerevisiae cells could withstand a subsequent higher dose of the same stress. Cross-adaptation also existed between the two stresses. Especially wild-type cells pretreated with 1%KCl (a hyperosmotic stress generating agent) and hyperosmotic-resistance mutant cells could significantly resist to lethal concentration of H2O2 (10mM). The addition of N-acetylcysteine (NAC) (30 mg 1-1) and MnSO4 (4mM) showed a strong reversion to hyperosmotic stress, which implicated that antioxidants participated in yeast adaptation to hyperosmosis. The two sublethal dose of stresses treatment, especially hyperosmosis, increased the level of GSH, CAT, SOD and total antioxidant capability (T-AOC) in yeast cells, which indicated adaptation between oxidative and hyperosmotic stresses was accompanied by the production of above several antioxidants. Furthermore, at least seven new proteins were induced by both oxidative and osmotic treatment.
Saccharomyces cerevisiae, Hyperosmotic stress, Oxidative stress, Protein oxidation, Adaptive response
-
36浏览
-
0点赞
-
0收藏
-
0分享
-
96下载
-
0评论
-
引用
杜连祥, Mei Guo, Fuping Lu, Jun Pu, Dongqing Bai, Lianxiang Du
Appl Microbiol Biotechnol (2005) 69: 178-183,-0001,():
-1年11月30日
A cDNA encoding for laccase was isolated from the ligninolytic fungus Trametes versicolor by RNA-PCR. The cDNA corresponds to the gene Lcc1, which encodes a laccase isoenzyme of 498 amino acid residues preceded by a 22-residue signal peptide. The Lcc1 cDNA was cloned into the vectors pMETA and pMETαA and expressed in Pichia methanolica. The laccase activity obtained with the Saccharomyces cerevisiae α-factor signal peptide was found to be twofold higher than that obtained with the native secretion signal peptide. The extracellular laccase activity in recombinants with the α-factor signal peptide was 9.79Uml−1. The presence of 0.2 mM copper was necessary for optimal activity of laccase. The expression level was favoured by lower cultivation temperature. The identity of the recombinant protein was further confirmed by immunodetection using Western blot analysis. As expected, the molecular mass of the mature laccase was 64.0kDa, similar to that of the native form.
-
31浏览
-
0点赞
-
0收藏
-
0分享
-
241下载
-
0评论
-
引用
【期刊论文】Purification and characterization of a novel fibrinolytic enzyme from Rhizopus chinensis 12
杜连祥, Liu Xiao-lan, Du Lian-xiang, Lu Fu-ping, Zheng Xi-qun, Xiao Jing
Appl Microbiol Biotechnol (2005) 67: 209-214,-0001,():
-1年11月30日
A novel fibrinolytic enzyme from Rhizopus chinensis 12 was purified through ammonium sulfate precipitation, hydrophobic interaction, ionic exchange, and gel filtration chromatography. The purification protocol resulted in a 893-fold purification of the enzyme, with a final yield of 42.6%. The apparent molecular weight of the enzyme was 18.0kDa, determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and 16.6kDa by gel filtration chromatography, which revealed a monomeric form of the enzyme. The isoelectric point of the enzyme estimated by isoelectric focusing electrophoresis was 8.5±0.1. The enzyme hydrolyzed fibrin. It cleaved the α, β, and γ chains of fibrinogen simultaneously, and it also hydrolyzed casein and N-succinyl-Ala-Ala-Pro-PhepNA. The enzyme had an optimal temperature of 45℃, and an optimal pH of 10.5. EDTA, PCMB, and PMSF inhibited the activity of the enzyme, and SBTI, Lys, TPCK, and Aprotinine had no obvious inhibition, which suggested that the activity center of the enzyme had hydrosulfuryl and metal. The first 12 amino acids of the N-terminal sequence of the enzyme were S-V-S-E-I-Q-LM-H-N-L-G and had no homology with that of other fibrinolytic enzyme from other microbes.
-
95浏览
-
0点赞
-
0收藏
-
0分享
-
91下载
-
0评论
-
引用
【期刊论文】Bioconversion of methyl-testosterone in a biphasic system
杜连祥, Bie Songtao *, Du lianxiang, Zhang Liming, Lu Fuping
Process Biochemistry 40(2005)3309-3313,-0001,():
-1年11月30日
The bioconversion of methyl-testosterone to methandienone in a biphasic system was studied using freeze-dried, thawed and growing Arthrobacter simplex AS 1.94* cells as biocatalyst. The biphasic system consists of an organic phase [30% (v/v)] with steroid 10g/L and an aqueous phase [70% (v/v)] containing the cells (5g.d.c.w/L). Carbon tetrachloride and Tween-80 were used as organic solvent and surfactant, respectively, and menadione was added as an external electron acceptor. The factors affecting the conversion rate in the two-phase system were investigated. The results showed that menadione was necessary in this system. Small amounts of product added to the reaction system can increase the dehydrogenation rate. Higher activities were obtained with thawed cells as compared to the freeze-dried and growing cells. Under the optimal operation conditions, more than 95% (w/w) of added methyl-testosterone could be converted to methandienone within 48h. Also, an easy way to recover the product in a high purity is proposed.
Methyl-testosterone, Methandienone, Bioconversion, Arthrobacter simplex, △1,, 2-Dehydrogenation, A biphasic system
-
36浏览
-
0点赞
-
0收藏
-
0分享
-
101下载
-
0评论
-
引用
杜连祥, Cai Heng *, Zhongjun Chen, Lianxiang Du & Fuping Lu
Biotechnology Letters (2005) 27: 1731-1736,-0001,():
-1年11月30日
Alpha amylase gene from Bacillus licheniformis was mutated by site-directed mutagenesis to improve its acid stability. The mutant gene was expression in Bacillus subtilis under the control of the promoter of sacB gene which was followed by either the a-amylase leader peptide of Bacillus licheniformis or the signal peptide sequence of sacB gene of Bacillus subtilis. Both peptides efficiently directed the secretion of a-amylase from the recombinant B. subtilis cells. The extracellular a-amylase activities in two recombinants were 1001 and 2012 U ml-1, respectively. The purity of the recombinant product was confirmed by SDSPAGE.
acid stability, a-amylase gene, Bacillus subtilis, expression
-
48浏览
-
0点赞
-
0收藏
-
0分享
-
55下载
-
0评论
-
引用
杜连祥, Zhongjun Chen, Heng Cai, Fuping Lu * & Lianxiang Du
Biotechnology Letters (2005) 27: 1745-1749,-0001,():
-1年11月30日
The expression of a synthetic gene encoding monellin, a sweet protein, in E. coli under the control of T7 promoter from phage is described. The single-chain monellin gene was designed based on the biased codons of E. coli so as to optimize its expression. Monellin was produced and accounted for 45% of total soluble proteins. It was purified to yield 43 mg protein per g dry cell wt. The purity of the recombinant protein was confirmed by SDS-PAGE.
Escherichia coli, expression, monellin, purification, sweet protein
-
67浏览
-
0点赞
-
0收藏
-
0分享
-
16下载
-
0评论
-
引用
【期刊论文】活酵母衍生物对丁抗氧化能力和部分免疫活性指标的影响*
杜连祥, 白东清, , 路福平, 王玉, 郭梅
动物学报,51 (4):664-668,2005,-0001,():
-1年11月30日
在饲料中添加0.4%活酵母衍生物(LYCD)喂养丁(Tinca tinca),对该鱼超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、氧化代谢产物丙二醛(MDA)以及溶菌酶(L YZ)等指标进行测定。结果表明:LYCD可显著提高机体的抗氧化能力;促进鱼体SOD酶的产生,试验与对照组鱼鳃SOD 酶活性表现出极显著差异,肝胰脏和脾脏SOD酶的含量显著提高;试验组肝胰脏CAT酶活性极显著升高而氧化代谢产物MDA 则极显著下降,脾脏也有显著差异;试验组鱼鳃和肝胰脏组织LYZ活性极显著高于对照组,脾脏L YZ 活性也显著提高,而肾脏没有明显变化;同时LYCD还能增加白细胞的吞噬能力,白细胞吞噬指数明显增加[动物学报51(4):664-668,2005] 。
活酵母衍生物, 丁, 免疫, 超氧化物歧化酶, 过氧化氢酶, 丙二醛, 溶菌酶
-
32浏览
-
0点赞
-
0收藏
-
0分享
-
52下载
-
0评论
-
引用
【期刊论文】甜蛋白Monellin 基因在大肠杆菌中的高效表达
杜连祥, 陈忠军, 蔡恒, 路福平*
生物工程学报,21(4):569~572,2005,-0001,():
-1年11月30日
根据已报道的单链monellin甜蛋白的氨基酸序列,采用细菌偏爱密码子,人工合成了全长294bp的monellin基因。插入到大肠杆菌表达载体pet-22b中,构建重组分泌型表达载体pETMO。经IPTG诱导pETMO所含有的甜蛋白基因可在大肠杆菌BL21(DE3)中高效表达,表达量占菌体可溶性蛋白的4418%。且经纯化后测定其甜度是蔗糖的3000倍。得到的甜蛋白热稳性及耐酸性均比天然产物有所提高。
甜蛋白Monellin, 细菌优化密码子, 重组PCR, 诱导表达
-
28浏览
-
0点赞
-
0收藏
-
0分享
-
57下载
-
0评论
-
引用
【期刊论文】人工合成的单链甜蛋白monellin基因在大肠杆菌中的高效表达
杜连祥, (陈忠军, 路福平, 蔡恒, 杜连祥)
食品与发西酵工业,2005,31(9):18~20,-0001,():
-1年11月30日
根据已报道的单链monellin甜蛋白的氨基酸序列,采用细菌偏爱密码子,人工合成了全长294bp的monellin基因。插入到大肠杆菌表达载体Pet-22b中,构建重组分泌型表达载体pETMO。经IPTG诱导pET-2MO所含有的甜蛋白基因可在大肠杆菌BL21(DE3)中高效表达,表达量占菌体可溶性蛋白的4418%。且经纯化后测定其甜度是蔗糖的3000倍。
甜蛋白monellin, 细菌优化密码子, 重组PCR, 诱导表达
-
30浏览
-
0点赞
-
0收藏
-
0分享
-
63下载
-
0评论
-
引用
【期刊论文】杂色云芝漆酶基因(Lccl)的克隆及在甲醇毕赤酵母中的表达
杜连祥, 郭梅, , 蒲军, 路福平*, 白东清
菌物学报,24(2):221~226,2005,-0001,():
-1年11月30日
以白腐菌杂色云芝Coriolus versicolor RNA为模板,通过RT-PCR获得漆酶Lccl基因的cDNA片段。构建了甲醇酵母表达质粒Pmeta-Lccl载体,并将其线性化后用电穿孔法导入Pichina methabolica PMAD16,部分阳性克隆的PCR结果表明Lccl基因已经整合到甲醇毕赤酵母染色体上,经摇瓶培养筛选出表达水平较高的酵母工程菌株。漆酶酶活力达53U/L
cDNA, 载体的构建, 异源表达
-
26浏览
-
0点赞
-
0收藏
-
0分享
-
31下载
-
0评论
-
引用