title: Biotransformation of menadione to its prenylated derivative MK-3 using recombinant Pichia pastoris.
abstract: Prenylated quinones, especially menaquinones, have significant physiological activities, but are arduous to synthesize efficiently. Due to the relaxed aromatic substrate specificity and prenylation regiospecificity at the ortho- site of the phenolic hydroxyl group, the aromatic prenyltransferase NovQ from Streptomyces may be useful in menaquinone synthesis from menadione. In this study, NovQ was overexpressed in Pichia pastoris. After fermentation optimization, NovQ production increased by 1617%. Then the different effects of metal ions, detergents and pH on the activity of purified NovQ were investigated to optimize the prenylation reaction. Finally, purified NovQ and cells containing NovQ were used for menadione prenylation in vitro and in vivo, respectively. Menaquinone-1 (MK-1) was detected as the only product in vitro with γ,γ-dimethylallyl pyrophosphate and menadione hydroquinol substrates. MK-3 at a concentration of 90.53 mg/L was detected as the major product of whole cell catalysis with 3-methyl-2-buten-1-ol and menadione hydroquinol substrates. This study realized whole cell catalysis converting menadione to menaquinones.
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Zhemin
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Genhai
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: The CAS Key Laboratory of Innate Immunity and Chronic Disease, Innovation Center for Cell Biology, School of Life Sciences, University of Science and Technology of China, Hefei, 230026, People's Republic of China.
authors.forename: Yugang
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Hefang
authors.initials: H
authors.lastname: Wu
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Sun
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Xihua
authors.initials: X
authors.lastname: Wu
authors.affiliation: Institute of Technical Biology and Agriculture Engineering, Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, Anhui, People's Republic of China. zhengzhiming2014@163.com.
authors.forename: Zhiming
authors.identifier: 0000-0002-0550-8087
authors.initials: Z
authors.lastname: Zheng
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D001711:Biotransformation; D002384:Catalysis; D004122:Dimethylallyltranstransferase / Q000378:metabolism; D045782:Hemiterpenes / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D060847:Metabolic Engineering; D000074041:Microorganisms, Genetically-Modified; D009943:Organophosphorus Compounds / Q000378:metabolism; D000439:Pentanols / Q000378:metabolism; D010843:Pichia / Q000201:enzymology*; D054876:Prenylation; D011994:Recombinant Proteins / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology; D013379:Substrate Specificity; D024482:Vitamin K 2 / Q000378:metabolism; D024483:Vitamin K 3 / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D045782:Hemiterpenes; D009943:Organophosphorus Compounds; D000439:Pentanols; D011994:Recombinant Proteins; D024482:Vitamin K 2; C043060:3,3-dimethylallyl pyrophosphate; C009034:prenol; D024483:Vitamin K 3; C544345:NovQ protein, Streptomyces; D004122:Dimethylallyltranstransferase
keywords: Aromatic prenyltransferase; Chemoenzymatic synthesis; Menadione prenylation; Pichia pastoris; Whole cell catalysis