Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana

Kajiura, Hiroyuki; Yamamoto, Kana; Misaki, Ryo; Fujiyama, Kazuhito

doi:https://doi.org/10.1111/pbi.70498

Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana

Kajiura, Hiroyuki; Yamamoto, Kana; Misaki, Ryo et al.

Plant Biotechnology Journal, 2025, 24(1), 187-203

Number of Access:101（2026-05-29 02:10 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/103816

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Title	Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana
Creator	NRID 1000030704209 1000030704209 ORCID 0000-0001-5811-1625 0000-0001-5811-1625 researchmap permalink 7000020437 7000020437 Scopus Author ID 16241653300 16241653300 CreatorName Kajiura, Hiroyuki
	CreatorName Yamamoto, Kana
	NRID 1000020571186 1000020571186 researchmap permalink read0090369 read0090369 Scopus Author ID 6603301567 6603301567 CreatorName Misaki, Ryo
	NRID 1000070209112 1000070209112 researchmap permalink read0014155 read0014155 Scopus Author ID 7005164172 7005164172 CreatorName Fujiyama, Kazuhito
Subject	apoplast
	Nicotiana benthamiana
	pathogen-related protein
	recombinant protein secretion
	signal peptide
Description	Kajiura H., Yamamoto K., Misaki R., et al. Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana. Plant Biotechnology Journal 24, 187 (2026); https://doi.org/10.1111/pbi.70498.
Abstract	Plants are promising next-generation hosts for recombinant protein production; however, major challenges remain with regard to enhancing the efficiency of downstream processing, particularly in the removal of cellular residues and purification of the expressed proteins. Strategies to overcome these limitations include targeting expressed recombinant proteins within a specific organelle or directing their secretion into the extracellular space, thereby facilitating purification by collecting the target matrix. In this study, we focused on protein secretion mechanisms and identified two pathogenesis-related proteins, glucan endo-1,3-β-glucosidase (GN) and chitinase 8 (Chi8), which accumulated in the apoplast washing fluid following Agrobacterium infiltration of Nicotiana benthamiana leaves. Both proteins contained signal peptides (SPs), SP𝙶𝙽 and SP𝙲𝚑𝚒𝟾, respectively. Although the intracellular accumulation of GFP was comparable regardless of the expression level, fusion with either SP𝙶𝙽 or SP𝙲𝚑𝚒𝟾 resulted in GFP accumulation within the apoplast. In contrast, in N. benthamiana, a mammalian-derived SP was less effective in facilitating GFP secretion than the plant-derived SPs. Additionally, replacing the SP of the mammalian-derived protein β-glucocerebrosidase (GCase) with SP𝙶𝙽 or SP𝙲𝚑𝚒𝟾 enhanced the secretion of GCase into the apoplast, indicating their applicability in protein production. Moreover, SP𝙶𝙽 and SP𝙲𝚑𝚒𝟾 directed the expressed proteins into the culture medium of N. benthamiana suspension cells. These results indicate that SP𝙶𝙽 and SP𝙲𝚑𝚒𝟾 function as effective secretion signals and highlight the potential application of endogenous SPs for enhancing recombinant protein production in plants.
Publisher	John Wiley and Sons Inc
Source Title	Plant Biotechnology Journal
Volume (Issue)	24(1)
Page	187-203
Date of Issued	2025-12-16
Language	English
Handle URL	https://hdl.handle.net/11094/103816
PISSN	14677644
eISSN	14677652
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1111/pbi.70498
Access Rights	open access
CopyRight	This article is licensed under a Creative Commons Attribution 4.0 International License.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Plant Biotechnology Journal. 2025, 24(1), p. 187-203
DC.title	Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana
DC.creator	Kajiura, Hiroyuki
	Yamamoto, Kana
	Misaki, Ryo
	Fujiyama, Kazuhito
DC.publisher	John Wiley and Sons Inc
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-12-16
DC.identifier	https://doi.org/10.1111/pbi.70498
DC.subject	apoplast
	Nicotiana benthamiana
	pathogen-related protein
	recombinant protein secretion
	signal peptide
DC.description	Kajiura H., Yamamoto K., Misaki R., et al. Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana. Plant Biotechnology Journal 24, 187 (2026); https://doi.org/10.1111/pbi.70498.
DCTERMS.abstract	Plants are promising next-generation hosts for recombinant protein production; however, major challenges remain with regard to enhancing the efficiency of downstream processing, particularly in the removal of cellular residues and purification of the expressed proteins. Strategies to overcome these limitations include targeting expressed recombinant proteins within a specific organelle or directing their secretion into the extracellular space, thereby facilitating purification by collecting the target matrix. In this study, we focused on protein secretion mechanisms and identified two pathogenesis-related proteins, glucan endo-1,3-β-glucosidase (GN) and chitinase 8 (Chi8), which accumulated in the apoplast washing fluid following Agrobacterium infiltration of Nicotiana benthamiana leaves. Both proteins contained signal peptides (SPs), SP𝙶𝙽 and SP𝙲𝚑𝚒𝟾, respectively. Although the intracellular accumulation of GFP was comparable regardless of the expression level, fusion with either SP𝙶𝙽 or SP𝙲𝚑𝚒𝟾 resulted in GFP accumulation within the apoplast. In contrast, in N. benthamiana, a mammalian-derived SP was less effective in facilitating GFP secretion than the plant-derived SPs. Additionally, replacing the SP of the mammalian-derived protein β-glucocerebrosidase (GCase) with SP𝙶𝙽 or SP𝙲𝚑𝚒𝟾 enhanced the secretion of GCase into the apoplast, indicating their applicability in protein production. Moreover, SP𝙶𝙽 and SP𝙲𝚑𝚒𝟾 directed the expressed proteins into the culture medium of N. benthamiana suspension cells. These results indicate that SP𝙶𝙽 and SP𝙲𝚑𝚒𝟾 function as effective secretion signals and highlight the potential application of endogenous SPs for enhancing recombinant protein production in plants.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	103816
citation_title	Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana
citation_author	Kajiura, Hiroyuki
	Yamamoto, Kana
	Misaki, Ryo
	Fujiyama, Kazuhito
citation_publisher	John Wiley and Sons Inc
citation_language	English
citation_date	2025-12-16
citation_journal_title	Plant Biotechnology Journal
citation_volume	24
citation_issue	1
citation_firstpage	187
citation_lastpage	203
citation_doi	https://doi.org/10.1111/pbi.70498
citation_public_url	https://hdl.handle.net/11094/103816

Protein-Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana

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