title: Fumaric acid production by Torulopsis glabrata: engineering the urea cycle and the purine nucleotide cycle.
abstract: A multi-vitamin auxotrophic Torulopsis glabrata strain, a pyruvate producer, was further engineered to produce fumaric acid. Using the genome-scale metabolic model iNX804 of T. glabrata, four fumaric acid biosynthetic pathways, involving the four cytosolic enzymes, argininosuccinate lyase (ASL), adenylosuccinate lyase (ADSL), fumarylacetoacetase (FAA), and fumarase (FUM1), were found. Athough single overexpression of each of the four enzymes in the cytosol improved fumaric acid production, the highest fumaric acid titer (5.62 g L(-1) ) was obtained with strain T.G-ASL(H) -ADSL(L) by controlling the strength of ASL at a high level and ADSL at a low level. In order to further improve the production of fumaric acid, the SpMAE1 gene encoding the C4 -dicarboxylic acids transporter was overexpressed in strain T.G-ASL(H) -ADSL(L) -SpMAE1 and the final fumaric acid titer increased to 8.83 g L(-1) . This study provides a novel strategy for fumaric acid biosynthesis by utilizing the urea cycle and the purine nucleotide cycle to enhance the bridge between carbon metabolism and nitrogen metabolism.
authors: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China; Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, 214122, China.
authors: Xiulai
authors: X
authors: Chen
authors: Jing
authors: J
authors: Wu
authors: Wei
authors: W
authors: Song
authors: Limei
authors: L
authors: Zhang
authors: Hongjiang
authors: H
authors: Wang
authors: Liming
authors: L
authors: Liu
mesh_terms: D000264:Adenylosuccinate Lyase / Q000235:genetics / Q000378:metabolism; D001123:Argininosuccinate Lyase / Q000235:genetics / Q000378:metabolism; D041221:Candida glabrata / Q000235:genetics / Q000378:metabolism*; D005650:Fumarates / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways; D011685:Purine Nucleotides / Q000378:metabolism*; D014508:Urea / Q000378:metabolism*
chemical_list: D005650:Fumarates; D005656:Fungal Proteins; D011685:Purine Nucleotides; C032005:fumaric acid; D014508:Urea; D001123:Argininosuccinate Lyase; D000264:Adenylosuccinate Lyase
keywords: Torulopsis glabrata; adenylosuccinate lyase; argininosuccinate lyase; fumaric acid; genome-scale metabolic model