title: Metabolic Engineering of Raoultella ornithinolytica BF60 for Production of 2,5-Furandicarboxylic Acid from 5-Hydroxymethylfurfural.
abstract: 2,5-Furandicarboxylic acid (FDCA) is an important renewable biotechnological building block because it serves as an environmentally friendly substitute for terephthalic acid in the production of polyesters. Currently, FDCA is produced mainly via chemical oxidation, which can cause severe environmental pollution. In this study, we developed an environmentally friendly process for the production of FDCA from 5-hydroxymethyl furfural (5-HMF) using a newly isolated strain, Raoultella ornithinolytica BF60. First, R. ornithinolytica BF60 was identified by screening and was isolated. Its maximal FDCA titer was 7.9 g/liter, and the maximal molar conversion ratio of 5-HMF to FDCA was 51.0% (mol/mol) under optimal conditions (100 mM 5-HMF, 45 g/liter whole-cell biocatalyst, 30°C, and 50 mM phosphate buffer [pH 8.0]). Next, dcaD, encoding dicarboxylic acid decarboxylase, was mutated to block FDCA degradation to furoic acid, thus increasing FDCA production to 9.2 g/liter. Subsequently, aldR, encoding aldehyde reductase, was mutated to prevent the catabolism of 5-HMF to HMF alcohol, further increasing the FDCA titer, to 11.3 g/liter. Finally, the gene encoding aldehyde dehydrogenase 1 was overexpressed. The FDCA titer increased to 13.9 g/liter, 1.7 times that of the wild-type strain, and the molar conversion ratio increased to 89.0%.
  
  
  
  In this work, we developed an ecofriendly bioprocess for green production of FDCA in engineered R. ornithinolytica This report provides a starting point for further metabolic engineering aimed at a process for industrial production of FDCA using R. ornithinolytica.
authors: School of Food Science and Technology, Jiangnan University, Wuxi, China.|Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors: Gazi Sakir
authors: GS
authors: Hossain
authors: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors: Haibo
authors: H
authors: Yuan
authors: Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors: Jianghua
authors: J
authors: Li
authors: School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
authors: Hyun-Dong
authors: HD
authors: Shin
authors: School of Food Science and Technology, Jiangnan University, Wuxi, China.
authors: Miao
authors: M
authors: Wang
authors: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors: Guocheng
authors: G
authors: Du
authors: Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors: Jian
authors: J
authors: Chen
authors: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China longliu@jiangnan.edu.cn.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors: Long
authors: L
authors: Liu
mesh_terms: D000080924:Aldehyde Dehydrogenase 1 Family; D000449:Aldehyde Reductase / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D018533:Biomass; D002262:Carboxy-Lyases / Q000235:genetics / Q000378:metabolism; D003998:Dicarboxylic Acids / Q000378:metabolism*; D004755:Enterobacteriaceae / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D005662:Furaldehyde / Q000031:analogs & derivatives* / Q000378:metabolism; D005663:Furans / Q000378:metabolism*; D007218:Industrial Microbiology / Q000379:methods; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways; D010084:Oxidation-Reduction; D011091:Polyesters / Q000737:chemistry; D050697:Retinal Dehydrogenase / Q000235:genetics / Q000378:metabolism
chemical_list: D003998:Dicarboxylic Acids; D005663:Furans; D007527:Isoenzymes; D011091:Polyesters; C008046:5-hydroxymethylfurfural; C551400:2,5-furandicarboxylic acid; D005662:Furaldehyde; D000449:Aldehyde Reductase; D000080924:Aldehyde Dehydrogenase 1 Family; D050697:Retinal Dehydrogenase; D002262:Carboxy-Lyases
keywords: 2,5-furandicarboxylic acid; 5-hydroxymethyl furfural; Raoultella ornithinolytica BF60; metabolic engineering; whole-cell biocatalyst