title: Cell-free synthesis of functional phospholipase A1 from Serratia sp.
abstract: BACKGROUND
  Phospholipase A1 is an enzyme that hydrolyzes phospholipids at the sn-1 position. It has potential applications across diverse fields including food, pharmaceutical, and biofuel industries. Although there has been increasing interest in the use of phospholipase A1 for degumming of plant oils during biodiesel production, production of recombinant phospholipase A1 has been hampered by low efficiency of gene expression and its toxicity to the host cell.
  
  
  RESULTS
  While expression of phospholipase A1 in Escherichia coli resulted in extremely low productivity associated with inhibition of transformed cell growth, drastically higher production of functional phospholipase A1 was achieved in a cell-free protein synthesis system where enzyme expression is decoupled from cell physiology. Compared with expression in E. coli, cell-free synthesis resulted in an over 1000-fold higher titer of functional phospholipase A1. Cell-free produced phospholipase A1 was also used for successfully degumming crude plant oil.
  
  
  CONCLUSIONS
  We demonstrate successful production of Serratia sp. phospholipase A1 in a cell-free protein synthesis system. Including the phospholipase A1 investigated in this study, many industrial enzymes can interfere with the regular physiology of cells, making cellular production of them problematic. With the experimental results presented herewith, we believe that cell-free protein synthesis will provide a viable option for rapid production of important industrial biocatalysts.
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764 Republic of Korea ; Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology, Daejeon, 305-600 Republic of Korea.
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Lim
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764 Republic of Korea.
authors.forename: Yu Jin
authors.initials: YJ
authors.lastname: Park
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764 Republic of Korea.
authors.forename: Yeon Jae
authors.initials: YJ
authors.lastname: Jang
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology, Daejeon, 305-600 Republic of Korea.
authors.forename: Ji Eun
authors.initials: JE
authors.lastname: Choi
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology, Daejeon, 305-600 Republic of Korea.
authors.forename: Joon Young
authors.initials: JY
authors.lastname: Oh
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology, Daejeon, 305-600 Republic of Korea.
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Park
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology, Daejeon, 305-600 Republic of Korea.
authors.forename: Jae Kwang
authors.initials: JK
authors.lastname: Song
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764 Republic of Korea.
authors.forename: Dong-Myung
authors.identifier: 0000-0001-7875-0694
authors.initials: DM
authors.lastname: Kim
keywords: Biodiesel; Cell-free protein synthesis; Enzymatic degumming; Industrial enzymes; Phospholipase A1