Food and Fermentation Industries >

2024 , Vol. 50 >Issue 7: 9 - 16

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035669

The construction of cell factory with high mannitol yield in Leuconostoc

  • LIU Chengchuan ,
  • JIN Hongxing
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  • (School of Chemical Engineering and Technology, Hebei University of Technology, Tianjin 300401, China)

Received date: 2023-04-02

  Revised date: 2023-05-08

  Online published: 2024-05-09

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Abstract

Mannitol is a hexasugar alcohol with many functions, which is widely used in medicine, food, chemical industry and electronics.In order to construct Leuconostoc with high yield of mannitol, the theory of transforming chassis cells into high-yield target product strain—the “source-sink” theory of cell factory was built.Through two homologous recombination, the operon and overlapping gene patterns were broken, and the mannitol efflux pump (MEP) gene sequence and the NADH-regenerating enzyme (formate dehydrogenase) gene sequence were site-direct inserted into the chromosome.When sucrose was 90 g/L, the mannitol yield of the strain that broke the operon and overlapping gene pattern of mep gene was 9.5% higher than that of the original strain (CGMCC1.10327).A strain with a copy of mep gene expression cassette was knocked into the chromosome of the original strain, and the substrate sucrose could be added to 120 g/L, and the output of mannitol reached 55.18 g/L.A strain with a copy of formate dehydrogenase gene expression cassette was knocked into the chromosome of CCTCC M2020762[Δdts1ΔldhΔpat::mdhΔstpk::mdhΔfk::mdhΔaldh::(mtld-mlp) 0385/0386::ldhA-eeΔadh::mepΔldh(0503)::mepΔldh(0373)::mep], and the substrate sucrose could be added to 145 g/L, and the output of mannitol reached 101.6 g/L.Make the cell factory have stronger “source” energy (NADH regeneration, etc.) and larger “sink” capacity (improve MEP activity), and obtain super-high yield of the target product.

Key words： cell factory; source-sink; Leuconostoc; mannitol; fermentation; synthetic biology

Cite this article

LIU Chengchuan , JIN Hongxing . The construction of cell factory with high mannitol yield in Leuconostoc[J]. Food and Fermentation Industries, 2024 , 50(7) : 9 -16 . DOI: 10.13995/j.cnki.11-1802/ts.035669

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