title: Heterologous Expression and Characterization of Mimosinase from Leucaena leucocephala.
abstract: Heterologous expression of eukaryotic genes in bacterial system is an important method in synthetic biology to characterize proteins. It is a widely used method, which can be sometimes quite challenging, as a number of factors that act along the path of expression of a transgene to mRNA, and mRNA to protein, can potentially affect the expression of a transgene in a heterologous system. Here, we describe a method for successful cloning and expression of mimosinase-encoding gene from Leucaena leucocephala (leucaena) in E. coli as the heterologous host. Mimosinase is an important enzyme especially in the context of metabolic engineering of plant secondary metabolite as it catalyzes the degradation of mimosine, which is a toxic secondary metabolite found in all Leucaena and Mimosa species. We also describe the methods used for characterization of the recombinant mimosinase.
authors: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, 1955 East West Road, Honolulu, HI, 96822, USA.
authors: Vishal Singh
authors: VS
authors: Negi
authors: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, 1955 East West Road, Honolulu, HI, 96822, USA. dulal@hawaii.edu.
authors: Dulal
authors: D
authors: Borthakur
mesh_terms: D003001:Cloning, Molecular; D004789:Enzyme Activation; D004798:Enzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007887:Fabaceae / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D008898:Mimosine / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA
chemical_list: D004798:Enzymes; D010940:Plant Proteins; D011994:Recombinant Proteins; D008898:Mimosine
keywords: 3-Hydroxy-4-pyridone; Aminotransferase; C-N lyase; Heterologous expression; Leucaena; Mimosinase; Mimosine; Secondary metabolite