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Production of human liver prolidase by Saccharomyces cerevisiae as host cells

Acta Pharmacologica Sinica 2004年第6期25卷 92-98页

作者： Shu-hao WANG, Min LIU, Mu-gen CHI, Qing-ding WANG, Man-ji SUN2 Institute of Pharmacologyand Toxicology, Academy of Military Medical Science, Beijing 100850, China Institute of Pharmacology and Toxicology Academy of Military Medical Science Beijing 100850 Chin

AIM: To clone and express the recombinant human liver prolidase in yeast and explore the activities of both dipeptidaseand organophosphoric acid anhydrolase(OPAA). METHODS:The cDNAencodinghuman liver prolidase derived from healthy adult liver was cloned into the pYES2, an expression vector of S cerevisiae, and then trans- formed into S cerevisiae INVSc1 by electroporation. The transformant with the highest enzymatic activity was induced by galactose for expression. The optimal induction conditions (temperature, induction time, and the initial amount of inoculation cells) were estimated by orthogonal experimental design. The recombinant prolidase and OPAA activities were assayed by spectrocolorimetric methods. RESULTS: The recombinant enzyme catalyzed the hydrolysis of organophosphorous compound soman as well as the hydrolysis of dipeptide Gly-Pro. Under the optimal induction conditions (20 h, 25 oC, initial OD600=0.4), the maximum activities of prolidase and OPAA came to 226.5 and 578 μmol·min-1·g-1 protein in cell lysate supernatants, respectively. SDS-PAGE of the recombinant enzyme in disrupted cell supernatants showed a molecular weight of 56 kDa. Intensity scanning of the SDS-PAGE gel revealed that the enzyme accounted for 3.16 % of the total protein in the supernatant. One liter incubation medium produced 7 g of wet yeast cell containing 4.56 mg of the recombination protein. CONCLUSION: The recombinant human liver prolidase produced by yeast cell (S cerevisiae) exhibited both dipeptidase and OPAA activities.

关键词： prolidase liver human S Cerevisiae organophosphorous compounds

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Assessment of the correlation between serum prolidase and alpha-fetoprotein levels in patients with hepatocellular carcinoma

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World Journal of Gastroenterology 2015年第22期21卷 6999-7007页

作者： Sevil Uygun Ilikhan Muammer Bilici Hatice Sahin Ayse Semra Demir Akca Murat Can Ibrahim Ilker Oz Berrak Guven M Cagatay Buyukuysal Yucel Ustundag Department of Internal medicine Bülent Ecevit University Faculty of Medicine Department of Family Medicine Bülent Ecevit University Faculty of Medicine Department of Biochemistry Bülent Ecevit University Faculty of Medicine Department of Radiology Bülent Ecevit University Faculty of Medicine Department of Biostatistic Bülent Ecevit University Faculty of Medicine Bülent Ecevit University Faculty of Medicine Department of Gastroenterology

AIM: To determine the predictive value of increased prolidase activity that reflects increased collagen turnover in patients with hepatocellular carcinoma(HCC).METHODS: Sixty-eight patients with HCC(mean age of 69.1 ± 10.1), 31 cirrhosis patients(mean age of59.3 ± 6.3) and 33 healthy volunteers(mean age of51.4 ± 12.6) were enrolled in this study. Univariate and multivariate analysis were used to evaluate the association of serum α-fetoprotein(AFP) values with HCC clinicopathological features, such as tumor size,number and presence of vascular and macrovascular invasion. The patients with HCC were divided into groups according to tumor size, number and presence of vascular invasion(diameters; ≤ 3 cm, 3-5 cmand ≥ 5 cm, number; 1, 2 and ≥ 3, macrovascular invasion; yes/no). Barcelona-clinic liver cancer(BCLC)criteria were used to stage HCC patients. Serum samples for measurement of prolidase and alphafetoprotein levels were kept at-80 ℃ until *** levels were measured spectrophotometrically and AFP concentrations were determined by a chemiluminescence immunometric commercial diagnostic ***: In patients with HCC, prolidase and AFP values were evaluated according to tumor size, number,presence of macrovascular invasion and BCLC staging classification. prolidase values were significantly higher in patients with HCC compared with controls(P prolidase levels were significantly associated with tumor size and number(P prolidase levels also differed in patients in terms of BCLC staging classification(P prolidase levels in HCC patients showed a significant difference compared with patients with cirrhosis(P prolidase and AFP values in

关键词： Alpha-fetoprotein Hepatocellular carcinoma prolidase Cirrhosis Macrovascular invasion

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