1.Optimization of liquid culture conditions of dihydropyrimidinase producing strain Pseudomonas putida 9801
Honghui HUANG ; Jun LI ; Zhuoyi HU
Chinese Journal of Biochemical Pharmaceutics 2001;22(1):15-17
Purpose The aim is to optimize the liquid culture conditions of dihydropyrimidinase producing strain Pseudomonas putida 9801. Methods Plackett-Burman design and spherical symmetric desig n were used.Results Optimum conditions for dihydropyrimidinas e formation of Pseudomonas putida 9801 were defined:yeast extract 2.39%, Glu cose 1.81%,Uracil 0.06%,K2HPO4*3H2O 0.2%, MgCl2*6H2O 0.05%and NaCl 0 .3%,when the strain was cultured at 32℃ for 10 h,about 3.02 units/ml of hydanto inase was obtained. This value was quite consistent with the theory value(2.91 u nits/ml).Conclusion The liquid culture conditions of dihydrop yrimidinase producing strain were optimized.
2.Expression and Purification of Hi-lys Peptide,a Recombinant Relevant to Hirudin
Jie WU ; Zhuoyi HU ; Jingjing LIU
Chinese Journal of Biochemistry and Molecular Biology 2005;21(3):287-291
A new fusion expression vector, pED-P8-Hi-lys was designed and constructed. It includes four parts, a 20 peptide sequence of hirudin that can maintain anticoagulant activity, the C-terminus of asparaginase as a fusion partner, basic octopeptide (KRKRKKSR) that makes the fusion partner easy to remove, and the unique acid-labile aspartyl-prolyl bond. It was transformed into E. coli BL-21 and the fusion protein (AnsB-C-P8-Hi-lys) was expressed effectively as inclusion bodies after inducing by lactose. The objective peptide Hi-lys was purified by means of cell disruption, washing, ethanol precipitation, acid hydrolysis, and DEAE-cellulose 52 column chromatography. The antithrombin activity of the purified Hi-lys peptide was about 50 ATU/mg by thrombin activity assays.
3.Cloning of D-hydantoinase Gene from Pseudomonas and Its Expression in E.coli
Zhiqiang LI ; Jingjing LIU ; Zhuoyi HU ; Zhenghua WANG ; Xin MING
Journal of China Pharmaceutical University 2001;(3):227-230
AIM The purpose is to construct D-hydantoinase genetic engineering strain for the purpose of the industrial production of D-p-hydroxyphenylglycine. METHODS D-hydantoinase gene was created from Pseudomonas putida 9801 by PCR technique and inserted into pMD18-T vector. The recombinant plasmid was transformed into several Escherichia coli strains. The positive transformants with D-hydantoinase activity were obtained by the two step screening, digoxigenin DNA labeling in situ hybridization and D-hydantoinase activity assay. RESULTS The D-hydantoinase activity of the genetic engineering strain E.coli BL21/pMD-dht was 1700 U*L-1 and increased as high as 8 times compared with those of wild-type strain Pseudomonas putida 9801. The subunit molecular weight of recombinant D-hydantoinase was about 53 kDa measured by SDS-PAGE. The amount of the recombinant D-hydantoinase was about 20 percent of total bacterial soluble proteins. CONCLUSION The genetic engineering strain E.coli BL21/pMD-dht possesses the initial industrial production prospects.
4.Review of Research on Issues and Countermeasures in the Early Warning System of Major Infectious Diseases Epidemic in China
Zhuoyi WANG ; Lei HAN ; Meirong HU ; Jinyi PAN ; Yuanlei YUE
Chinese Medical Ethics 2023;36(11):1231-1237
Major infectious disease epidemic continues to pose a threat to human health and society, and the effective establishment and implementation of an early warning system plays a key role in addressing public health security risks. At present, the research on early warning of infectious diseases in the academic community mainly focuses on early warning information system, early warning mechanism, laws and regulations of early warning of infectious diseases, and some studies lack specific suggestions on operation methods. By collating and summarizing the literature from 2002 to 2022, regarding the early warning system and mechanism of major infectious diseases, this paper focused on analyzing the public health ethical dilemmas existing in the early warning process and discussing how to strengthen the construction of the early warning system of infectious diseases, so as to lay the foundation for creating more scientific early warning schemes of infectious diseases.
5.Regulation of aerobic glycolysis to decelerate tumor proliferation by small molecule inhibitors targeting glucose transporters.
Meng GAO ; Jian HUANG ; Xin JIANG ; Yafei YUAN ; Huanhuan PANG ; Shuchen LUO ; Nan WANG ; Chengbo YAO ; Zuwan LIN ; Debing PU ; Shuo ZHANG ; Pengcheng SUN ; Zhuoyi LIU ; Yu XIAO ; Qian WANG ; Zeping HU ; Hang YIN
Protein & Cell 2020;11(6):446-451