王正祥
1、工业微生物资源和育种;2、酶的分子结构、功能、人工进化和高效表达;3、分子微生物学。
个性化签名
- 姓名:王正祥
- 目前身份:
- 担任导师情况:
- 学位:
-
学术头衔:
博士生导师, 教育部“新世纪优秀人才支持计划”入选者
- 职称:-
-
学科领域:
发酵工程
- 研究兴趣:1、工业微生物资源和育种;2、酶的分子结构、功能、人工进化和高效表达;3、分子微生物学。
王正祥,男,1964年3月生,江苏盐城市人,民盟、中共党员,工学博士,博士后(Westfalische Wilhelms University-Muenster分子微生物学和生物工程研究所2001.3-2002.10),访问研究教授(Stellenbosch大学微生物学系2000.1-2000.12)。现为江南大学教授、博导,教育部新世纪优秀人才支持计划人选,江苏省高校新世纪学术带头人培养人选(2000),江南大学生物工程学科学术带头人之一,《生化与分子生物学》学科带头人,《工业微生物学》学术带头人。主要参与了“好氧发酵法生产甘油”(国家“九五”科技攻关项目)的研究。主持完成了“乙醇基因工程生产菌的构建”(教育部科技司)的研究和江苏省自然科学基金项目1项,江苏省教委自然科学基金项目1项,江苏省骨干教师基金项目1项。主要参与完成其他省级项目3项。共发表研究论文107篇,SCI级论文7篇;副主编著作2部,申报国家发明专利2项。正负责“新型生物饲料关键技术研究与新产品开发”(国家863计划),主持“工业微生物资源数据库”(教育部)和“利用现代生物技术改造产酶工业生产菌种”(企业委托)的研究。曾获江苏省有突出贡献的中青年专家(1995),江苏省“八五”先进科技工作者(1995)和扬州市优秀大中专毕业生(1996)等荣誉。现主要研究方向和研究兴趣为:1、工业微生物资源和育种;2、酶的分子结构、功能、人工进化和高效表达;3、分子微生物学。
-
主页访问
4058
-
关注数
0
-
成果阅读
477
-
成果数
9
【期刊论文】耐高渗压高产甘油的一个假丝酵母新种——产甘油假丝酵母*
王正祥, 诸葛健, 方慧英
微生物学报,1999,39(1):68~74,-0001,():
-1年11月30日
从自然标本中分离获得一高产甘油的菌株WL2002-5,仅发酵葡萄糖及微发酵蔗糖,能利用葡萄糖、蔗糖、乙醇生长,微利用甘油和柠檬酸,不利用肌醇、硝酸盐、赤藓醇、阿拉伯醇、甘露醇,与DBB显色反应为阴性,可在含500g/L葡萄糖或100mL/L醋酸的培养基中40℃下生长,可在水活度为0.890-0.900的培养基中生长,出芽生殖,易形成“假丝酵母菌型”假菌丝,不进行有性生殖,线粒体DNA的分子量为20kb,是假丝酵母属的一个新种,定名为产甘油假丝酵母(Candida glycerol genesis Zhuge sp. nov.)。
产甘油假丝酵母, 鉴定, 发酵
-
84浏览
-
0点赞
-
0收藏
-
0分享
-
201下载
-
0评论
-
引用
【期刊论文】利用木糖和葡萄糖合成乙醇的新型重组大肠杆菌的研究
王正祥, 孙金凤, 徐敏, 张峰, 王正祥*
微生物学报,2004,44(5):600~604,-0001,():
-1年11月30日
利用PCR方法从运动发酵单孢菌染色体DNA扩增出乙醇合成途径的关键酶基因pdc、adhB,分别用tac启动子控制表达,构建了可以在Escherichia coli JMI 09中表达的重组质粒pKK-PA、pEtac-PA。初步的乙醇发酵结果表明,在 E. coli中只引入 adhB基因不能拓宽其中的产乙醇途径,引入pdc基因可以与宿主自身的ADH酶协同作用,使碳流有效导向产乙醇方向。同时引入pdc、adhB基因可以在宿主E. coli中成功建立产乙醇途径。
重组大肠杆菌, 乙醇, 发酵
-
44浏览
-
0点赞
-
0收藏
-
0分享
-
209下载
-
0评论
-
引用
王正祥, 诸葛健, 曹钰, 陈珺, 方慧英
微生物学报,2000,40(2):180~187,-0001,():
-1年11月30日
本文对产甘油假丝酵母的甘油代谢关键酶进行了研究,发现产甘油假丝酵母同化甘油能力极弱,少量葡萄糖明显改善其同化甘油的能力;线粒体3-磷酸甘油脱氢酶受3-磷酸甘油的强烈诱导,受葡萄糖代谢的阻遏。在甘油发酵过程中,产甘油假丝酵母胞浆3-磷酸甘油脱氢酶酶活处于较高水平并在36h和60h时出现两次酶活高峰,其中第一次酶活峰值水平决定产甘油假丝酵母的甘油合成和积累水平,成为甘油高速积累期(18~48h)甘油合成的关键 性的限速酶。在甘油发酵18~48h内,3-磷酸甘油酯酶的酶活处于高水平,并在36h时出现酶活峰值;处于缓慢甘油积累阶段的48~72h间,3-磷酸甘油酯酶已处于低水平表达,此时,3-磷酸甘油酯酶则成为甘油合成的限速酶。 产甘油假丝酵母稳定并高表达其胞浆3-磷酸甘油脱氢酶基因并且其所表达的3-磷酸甘油酯酶酶活远高于胞浆3-磷酸甘油脱氢酶这一特征是其高产甘油根本所在。
甘油, 产甘油假丝酵母, 线粒体3-磷酸甘油脱氢酶, 胞浆3-磷酸甘油脱氢酶, 3-磷酸甘油酯酶
-
77浏览
-
0点赞
-
0收藏
-
0分享
-
238下载
-
0评论
-
引用
【期刊论文】产甘油假丝酵母胞浆32磷酸甘油脱氢酶编码基因的克隆·
王正祥, 诸葛健
微生物学报,1999,39(4):321~326,-0001,():
-1年11月30日
当酵母细胞处于高渗压环境时,甘油被诱导合成以提高其胞内渗透压,这一过程受HOG途径的调控。GPDI基因为HOG途径的重要靶基因,高效表达使胞内3-磷酸甘油脱氢酶酶活水平提高可极大地提高甘油的产量。本研究将产甘油假丝酵母(Candida glycerolo-genesis)染色体DNA经Sau3 AI部分酶解后的5-10kbDNA片段与经Bam HI线性化及CIP处理过的酵母-大肠杆菌穿梭质粒YEp51连接,以大肠杆菌DH5a为受体,构建产甘油假丝酵母的染色体基因文库。通过遗传互补法,在含50g/L氯化钠的培养基上筛选出15个转化子,对转化子0601进行了进一步鉴定,转化子0601所含质粒YEp0601带有YEp51的标记并可以消除Saccbaro myces cervisiae642菌株由于其GPD1,GAPD2两基因的缺失突变而表现出的渗透压敏感性,表明已克隆到产甘油假丝酵母的编码胞浆3-磷酸甘油脱氢酶的基因。
产甘油假丝酵母, 3-磷酸甘油脱氢酶基因, 克隆
-
46浏览
-
0点赞
-
0收藏
-
0分享
-
121下载
-
0评论
-
引用
【期刊论文】Two phenotypically compensating isocitrate dehydrogenases inRalstonia eutropha
王正祥, Zheng-Xiang Wang a;b, Christian Bra
FEMSM icrobiology Letters 11174(2003)1-8,-0001,():
-1年11月30日
The tricarboxylic acid (TCA) cycle enzyme isocitrate dehydrogenase (IDH) and the glyoxylate bypass enzyme isocitrate lyase are involved in catabolism of isocitrate and play a key role in controlling the metabolic flux between the TCA cycle and the glyoxylate shunt. Two IDH genes icd1 and icd2 of Ralstonia eutropha HF39, encoding isocitrate dehydrogenase 1 (IDH1) and isocitrate dehydrogenase 2 (IDH2), were identified and characterized. Icd1 was functionally expressed in Escherichia coli, whereas icd2 was expressed in E. coli but no activity was obtained. Interposon-mutants of icd1 (HF39vicd1) and icd2 (HF39vicd2) of R. eutropha HF39 were constructed and their phenotypes were investigated. HF39vicd1 retained 43% of IDH activity, which was not induced by a cetate, and HF39vicd2 expressed 74% of acetate-induced IDH activity. Both HF39vicd1and HF39vicd2 kept the same growth rate on acetate as the wild-type. These data suggested that IDH1 is induced by acetate. The interposon-mutants HF39vicd1 and HF39vicd2 accumulated the same amount of poly(3-hydroxybutyric acid) as the wild-type.
Isocitrate dehydrogenase, Isocitrate lyase, Malate synthase, Polyhydroxyalkanoic acids, Ralstonia eutropha
-
41浏览
-
0点赞
-
0收藏
-
0分享
-
124下载
-
0评论
-
引用
【期刊论文】The glyoxylate bypass of Ralstonia eutropha
王正祥, Zheng-Xiang Wang a;b, Christian O. Bramer a, Alexander Steinbuchel a;
FEMS Microbiology Letters 228(2003)63-71,-0001,():
-1年11月30日
The glyoxylate bypass genes aceA1 (isocitrate lyase 1, ICL1), aceA2 (isocitrate lyase 2, ICL2) and aceB1 (malate synthase, MS1) of Ralstonia eutropha HF39 were cloned, sequenced and functionally expressed in Escherichia coli. Interposon-mutants of all three genes (vaceA1, vaceA2 and vaceB1) were constructed, and the phenotypes of the respective mutants were investigated. Whereas R. eutropha HF39vaceA1 retained only 19% of ICL activity and failed to grow on acetate, R. eutropha HF39vaceA2 retained 84% of acetate-inducible ICL activity, and growth on acetate was not retarded. These data suggested that ICL1 is in contrast to ICL2 induced by acetate and specific for the glyoxylate cycle. R. eutropha HF39vaceB1 retained on acetate as well as on gluconate about 41-42% of MS activity and exhibited retarded growth on acetate, indicating the presence of a second hitherto not identified MS in R. eutropha HF39. Whereas in R. eutropha HF39vaceA1 and R. eutropha HF39vaceA2 the yields of poly(3-hydroxybutyric acid), using gluconate as carbon source, were significantly reduced, R. eutropha HF39vaceB1 accumulated the same amount of this polyester from gluconate as well as from acetate as R. eutropha HF39.
Glyoxylate bypass, Isocitrate lyase, Malate synthase, Polyhydroxyalkanoate, Ralstonia eutropha
-
56浏览
-
0点赞
-
0收藏
-
0分享
-
56下载
-
0评论
-
引用
【期刊论文】Glycerol production by a novel osmotolerant yeast Candida glycerinogenes
王正祥, J. Zhuge, H.-Y. Fang, Z.-X. Wang, D.-Z. Chen H. -R. Jin, H. -L. Gu
Appl Microbiol Biotechnol (2001) 55: 686-692,-0001,():
-1年11月30日
Candida glycerinogenes, an osmotolerant yeast isolated from a natural sample in an environment of high osmotic pressure, had a modest sugar-tolerance and an extremely high glycerol productivity. The optimum conditions for glycerol formation by C. glycerinogenes were a temperature of 29-33℃ and a pH of 4-6. The optimum medium for glycerol production consisted of 230-250g glucose/l, 2g urea/l and 5ml corn steep liquor/l (55-65mg phosphates/l); the pH was not adjusted. The highest yield of glycerol was 64.5% (w/w) based on consumed glucose from 240g glucose/l, and the highest concentration of glycerol was 137g/l from 260g glucose/l. These results were obtained by using a 30-l agitated fermentor under optimal fermentation conditions. In ten batch-fermentations carried out in a 50,000-l airlift fermentor, an average yield of glycerol of 50.67% (w/w) and an average glycerol concentration of 121.9g/l were obtained from an average 240.6g glucose/l.
-
52浏览
-
0点赞
-
0收藏
-
0分享
-
485下载
-
0评论
-
引用
-
33浏览
-
0点赞
-
0收藏
-
0分享
-
91下载
-
0评论
-
引用
王正祥, Zheng-Xiang Wang, , Gerald Kayingo, Anders Blomberg and Bernard A. Prior*
Yeast 2002; 19: 1447-1458.,-0001,():
-1年11月30日
The dihydroxyacetone pathway, an alternative pathway for the dissimilation of glycerol via reduction by glycerol dehydrogenase and subsequent phosphorylation by dihydroxyacetone (DHA) kinase, is activated in the yeasts Saccharomyces cerevisiae and Zygosaccharomyces rouxii during osmotic stress. In experiments aimed at investigating the physiological function of the DHA pathway in Z. rouxii, a typical osmotolerant yeast, we cloned and characterized a DAK gene encoding dihydroxyacetone kinase from Z. rouxii NRRL 2547. Sequence analysis revealed a 1761 bp open reading frame, encoding a peptide composed of 587 deduced amino acids with the predicted molecular weight of 61 664 Da. As the amino acid sequence was most closely homologous (68% identity) to the S. cerevisiae Dak1p, we named the gene and protein ZrDAK1 and ZrDak1p, respectively. A putative ATP binding site was also found but no consensus element associated with osmoregulation was found in the upstream region of the ZrDAK1 gene. The ZrDAK1 gene complemented a S. cerevisiae W303-1A dak1 dak2 strain by improving the growth of the mutant on 50mmol/l dihydroxyacetone and by increasing the tolerance to dihydroxyacetone in a medium containing 5% sodium chloride, suggesting that it is a functional homologue of the S. cerevisiae DAK1. However, expression of the ZrDAK1 gene in the S. cerevisiae dak1 dak2 strain had no significant effect on glycerol levels during osmotic stress. The ZrDAK1 sequence has been deposited in the public data bases under Accession No.AJ294719; regions upstream and downstream of ZrDAK1are deposited as Accession Nos AJ294739 and AJ294720, respectively. Copyright 2002 John Wiley & Sons, Ltd. Copyright
Zygosaccharomyces rouxii, dihydroxyacetone, osmotic stress, DAK1, DHA pathway
-
44浏览
-
0点赞
-
0收藏
-
0分享
-
77下载
-
0评论
-
引用