title: Production of β-carotene by expressing a heterologous multifunctional carotene synthase in Yarrowia lipolytica.
abstract: OBJECTIVE
  To obtain functional expression of a heterologous multifunctional carotene synthase containing phytoene synthase, phytoene dehydrogenase, and lycopene β-cyclase activities encoded by carS from Schizochytrium sp. in order to allow Yarrowia lipolytica to produce β-carotene.
  
  
  RESULTS
  To increase the integration efficiency of a 3.8 kb carS under the control of PGPDpromoter with a 2 kb selection marker, ura3, along with a geranylgeranyl diphosphate synthase (GGS1) expression cassette (~10 kb in total), was inserted into the Y. lipolytica chromosome, and the DNA assembler method was combined with double chromosomal deletions of ku70 and ku80. This method resulted in a 13.4-fold increase in integration efficiency compared with the original method, reaching 63% (10/16). The resulting recombinant Y. lipolytica produced 0.41 mg β-carotene per g dry cell weight, while the wild type did not produce any indicating the functionality of the multifunctional carotene synthase in Y. lipolytica.
  
  
  CONCLUSIONS
  Expression of GGS1 and a multifunctional carotene synthase from Schizochytrium sp. in Y. lipolytica led to β-carotene production. DNA assembler efficiency was greatly increased by the deletion of ku70 and ku80, which resulted in decreased in vivo nonhomologous end-joining (NHEJ) in Y. lipolytica.
authors.affiliation: School of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
authors.forename: Shuliang
authors.initials: S
authors.lastname: Gao
authors.affiliation: College of Life and Environmental Sciences, Shanghai Normal University, 100 Guilin Rd, Shanghai, 200234, China.
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Tong
authors.affiliation: Shanghai Laiyi Center for Biopharmaceutical R&D, 800 Dongchuan Road, Shanghai, 200240, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Shanghai Laiyi Center for Biopharmaceutical R&D, 800 Dongchuan Road, Shanghai, 200240, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Ge
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China.|Shanghai Research and Development Center of Industrial Biotechnology, 528 Ruiqing Road, Shanghai, 201201, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: School of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. hccb001@163.com.|Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Road, Shanghai, 200040, China. hccb001@163.com.
authors.forename: Daijie
authors.initials: D
authors.lastname: Chen
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China.|Shanghai Research and Development Center of Industrial Biotechnology, 528 Ruiqing Road, Shanghai, 201201, China.|Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), 200 North Zhongshan Road, Nanjing, 211816, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
mesh_terms: D019149:Bioreactors; D004798:Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058009:Stramenopiles / Q000201:enzymology / Q000235:genetics; D025062:Yarrowia / Q000235:genetics* / Q000378:metabolism; D019207:beta Carotene / Q000032:analysis / Q000378:metabolism*
chemical_list: D004798:Enzymes; D011994:Recombinant Proteins; D019207:beta Carotene
keywords: Carotene synthase complex; DNA assembler; Multifunctional carotene synthase; Yarrowia lipolytica; carS; β-Carotene