Biochemical characterization of a novel L - asparaginase with low 1 glutaminase activity from Rhizomucor miehei and its application in 2 food safety and leukemia treatment

Biochemical characterization of a novel L-asparaginase with low 1 glutaminase activity from Rhizomucor miehei and its application in 2 food safety and leukemia treatment 3 Linhua Huang, Yu Liu, Yan Sun, Qiaojuan Yan , Zhengqiang Jiang #, * 4 Bioresource Utilization Laboratory, College of Engineering, China Agricultural 5 University, Beijing 100083, China; School of Biological Science and Medical 6 Engineering, Beihang University, Beijing 100191, China; Department of 7 Biotechnology, College of Food Science and Nutritional Engineering, China 8 Agricultural University, Beijing 100083, China 9 Corresponding author. Tel.: +86 10 62737689; fax: +86 10 82388508. E-mail addresses: 10 [email protected] (Q.J. Yan); [email protected] (Z.Q. Jiang). 11 *Address for correspondence: Professor Dr. Zhengqiang Jiang 12 Post Box 294, China Agricultural University, Beijing 100083, China. Tel.: +86 10 13 62737689; fax: +86 10 82388508. E-mail addresses: [email protected] (Z.Q. Jiang). 14 15 Running title: Fungal L-asparaginase and its applications 16 Abbreviations used: AML, Astragalus membranaceus lectin; CHES, 17 2-(cyclohexylamino) ethanesulfonic acid; IPTG, isopropyl β-D-thiogalactopyranoside; 18 MES, 2-(N-morpholino) ethane sulfonic acid; MTT, 19 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; ORF, open reading 20 frame; PCR, polymerase chain reaction; PI, Prodium Iodide; RACE, rapid amplification 21 of cDNA ends; RmAsnase, Rhizomucor miehei L-asparaginase. 22 AEM Accepts, published online ahead of print on 20 December 2013 Appl. Environ. Microbiol. doi:10.1128/AEM.03523-13 Copyright © 2013, American Society for Microbiology. All Rights Reserved.