Please use this identifier to cite or link to this item:
Title: The development of human recombinant arginase as a novel agent in the treatment of human cancer
Authors: Tsui, Sam-mui
Keywords: Hong Kong Polytechnic University -- Dissertations
Cancer -- Treatment
Issue Date: 2004
Publisher: The Hong Kong Polytechnic University
Abstract: Some tumour cells have been shown to be auxotrophic for arginine. Enzyme degradation of extracellular arginine becomes a possible mean for inhibiting tumour growth but harmless to normal cells. Arginase is one of the arginine depriving enzymes that catalyses the hydrolysis of arginine to urea and ornithine. Arginase was shown by others to have anti-tumour activity and is potentially useful for treatment of cancer. In 1990, recombinant human arginase was produced successfully from an Escherichia coli expression system. However, E. coli is pathogenic and the yield of arginase was low. To overcome these problems, we produced recombinant human arginase using a non-pathogenic prophage-based Bacillus subtilis expression system. In addition, the arginase was tagged with 6 histidines at the N-terminus to allow single-step purification, by which about 100 mg of arginase was obtained from 1 L of fermentation culture in unprecedented high purity as well as activity. In phosphate-buffered saline buffer (pH 7.4), the purified enzyme is a dimer with Km and kcat values of 1.89 mM and 1.80 s⁻¹, respectively, and the specific activity is 500-600 I.U. mg⁻¹. The blood circulation half-life of the recombinant arginase was found to be too short and was not effective to deplete serum arginine. To overcome this problem, the enzyme was modified with polyethylene glycol (PEG) to produce pegylated arginase, arginase-SPA-PEG₅₀₀₀. Arginase-SPA-PEG₅₀₀₀ is fully active, with substantially increased circulation half-life, and retains all the enzyme kinetic parameters as the native arginase. When given a single intraperitoneal (i.p.) injection of 1,500 I.U. of arginase-SPA-PEG₅₀₀₀ to rats, the plasma arginine level was decreased to a non-detectable level for about six days. These data indicate that the pegylated human arginase, arginase-SPA-PEG₅₀₀₀, is safe and could be further developed to function as an effective arginine depriving anti-cancer drug.
Description: vi, 166 leaves : ill. ; 30 cm.
PolyU Library Call No.: [THS] LG51 .H577M ABCT 2004 Tsui
Rights: All rights reserved.
Appears in Collections:Thesis

Files in This Item:
File Description SizeFormat 
b20593272_link.htmFor PolyU Users 162 BHTMLView/Open
b20593272_ir.pdfFor All Users (Non-printable)2.94 MBAdobe PDFView/Open
Show full item record
PIRA download icon_1.1View/Download Contents

Page view(s)

Last Week
Last month
Citations as of Dec 10, 2018


Citations as of Dec 10, 2018

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.