Evaluation of the function of a luciferase-like monooxygenase homologue in 4,4´-dithiodibutyric acid catabolism in Rhodococcus erythropolis MI2

作     者：M.Venkateswar Reddy Alexander Steinbüchel 

作者机构：Institut für Molekulare Mikrobiologie und BiotechnologieWestfälische Wilhelms-Universität Münster48149 MünsterGermany Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic Institute1108th StreetTroyNY 12180USA Environmental Sciences DepartmentKing Abdulaziz UniversityJeddahSaudi Arabia 

出 版 物：《Systems Microbiology and Biomanufacturing》 (系统微生物学与生物制造（英文）)

年 卷 期：2022年第2卷第3期

页      面：523-532页

核心收录：

学科分类：08[工学] 082701[工学-核能科学与工程] 0827[工学-核科学与技术] 

基　　金：supported by Alexander von Humboldt(AvH)foundation Germany(Ref No:IND 1162665 HFST-P) 

主　　题：Deletion mutant Monooxygenase Polythioesters Protein purification Rhodococcus erythropolis MI2 4,4´-Dithiodibutyric acid(DTDB) 

摘      要：The bacterium Rhodococcus erythropolis MI2 uses 4,4´-dithiodibutyric acid(DTDB)as carbon source to synthesize polythioesters(PTE).The first step for the production of PTE using DTDB is catalyzed by an NADH:flavin oxidoreductase(nox)as it was previously shown in our laboratory,and the second step is catabolized by a putative luciferase-like monooxygenase(Llm).In the current study,experiments were carried out to identify the function of ***,the llm gene,which encodes for the Llm protein,was amplified from the genomic DNA of MI2 using polymerase chain reaction and expressed in Escherichia coli BL21 *** purification was done using His Spin Trap affinity *** assay was carried out using the purified protein and p-coumaric acid as substrate giving a specific activity of 1.6 U/mg protein for the purified *** responsible gene(llm)was deleted in the genome of MI2,and a single deletion mutant was subsequently ***,growth of the wild-type strain(MI2)and the mutant strain(MI2Δllm)were compared using DTDB or succinate as carbon *** the wild type was successfully grown with DTDB or succinate,the llm-negative mutant exhibited low grow with DTDB although it grows very well with succinate.