title: Metabolic engineering of Bacillus subtilis for the efficient biosynthesis of uniform hyaluronic acid with controlled molecular weights.
abstract: Bacillus subtilis was engineered into an efficient hyaluronic acid (HA) producer by introducing two inducible artificial operons carrying HA synthase gene from Pasteurella multocida and precursor genes encoding enzymes involved in synthesis of the sugar precursors. A two-stage induction strategy was established for metabolic engineering of recombinant B. subtilis to efficiently produce uniform HA with controlled molecular weights. Strain TPG223 produced larger HA molecules (yield=6.8 g/L; molecular weight=4.5 MDa) than strain PG6181 (yield=2.4 g/L; molecular weight=13 KDa), indicating that the enzymes involved in the synthesis of UDP-glucuronic acid are essential for HA biosynthesis. Strain TPG223 was able to synthesize HA molecules ranging in molecular weight from 8 KDa to 5.4 MDa indicating that size control is achievable in vivo through appropriate tools. The work reported here not only advanced mechanisms research of size control in vivo, but also could be an attractive alternative for commercial preparation of uniform size-defined HA.
authors: State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China.
authors: Yuning
authors: Y
authors: Jia
authors: Jing
authors: J
authors: Zhu
authors: Xiaofei
authors: X
authors: Chen
authors: Dongyang
authors: D
authors: Tang
authors: Ding
authors: D
authors: Su
authors: Wenbing
authors: W
authors: Yao
authors: Xiangdong
authors: X
authors: Gao
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D014453:Glucuronosyltransferase / Q000235:genetics*; D000076002:Hyaluronan Synthases; D006820:Hyaluronic Acid / Q000096:biosynthesis* / Q000737:chemistry*; D060847:Metabolic Engineering / Q000379:methods*; D009876:Operon / Q000235:genetics; D016979:Pasteurella multocida / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D006820:Hyaluronic Acid; D014453:Glucuronosyltransferase; D000076002:Hyaluronan Synthases