Region-based segmental swapping of homologous enzymes for higher cadaverine production
Kim, Seungjin (Pohang University of Science and Technology. Department of Chemical Engineering)
Ye, Dae-Yeol (Pohang University of Science and Technology. Department of Chemical Engineering)
Lim, Hyun Gyu (Inha University. Department of Biological Sciences and Bioengineering)
Noh, Myung Hyun (Korea Research Institute of Chemical Technology. Research Center for Bio-based Chemistry)
Yang, Jae-Seong (Centre de Recerca en Agrigenòmica)
Jung, Gyoo Yeol (Pohang University of Science and Technology. School of Interdisciplinary Bioscience and Bioengineering)

Date:	2025
Abstract:	Background: Cadaverine, displaying potential in medicine, agriculture and polyamide production, is biologically produced through L-lysine decarboxylation. Considering the potential of the polyamide market, its biological production has been focused on with following diverse efforts to improve the production. In Escherichia coli, lysine decarboxylase exists in two forms: CadA and LdcC, and it is known that CadA exhibits superior catalytic activity compared to LdcC. Despite its potential, cadaverine production is limited due to increased intracellular pH, which destabilizes the decameric structure of CadA and inhibits its activity. Results: In this study, based on the structural analysis, a chimeric CadA enzyme, CL2, was engineered by replacing its pH-sensitive region with a structurally stable counterpart derived from LdcC. The resulting BLCL2 strain with CL2 produced 1. 12 g/L of cadaverine-1. 96 times higher than BLC strain with the wild type CadA in flask culture. Compared to the wild type CadA, structural modifications enhanced pH stability and improved the affinity of CadA toward pyridoxal 5-phosphate (PLP), its cofactor. Conclusions: This study developed the improved strains for cadaverine production by creating the new enzyme, which is validated by enhanced amount of cadaverine. In addition, the segmental swapping guided by structure analysis was exhibited as the one of effective method in protein engineering strategies. These advancements offer a promising approach to optimizing cadaverine biosynthesis for industrial applications.
Grants:	Agencia Estatal de Investigación RYC2020-028880-I
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Cadaverine ; Chimeric enzyme ; Lysine decarboxylase ; pH stability ; Cofactor affinity
Published in:	Microbial cell factories, Vol. 24, Num. 1 (December 2025) , art. 120, ISSN 1475-2859

DOI: 10.1186/s12934-025-02739-4
PMID: 40405206

11 p, 2.0 MB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > CRAG (Centre for Research in Agricultural Genomics)
Articles > Research articles
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