Effective Repeated Production of γ-glutamylcysteine, Essential For Intracellular Glutathione Production, Using Cellulose-immobilized Phytochelatin Synthase-like Enzyme NsPCS

Nakai, Takuya; Hirata, Kazumasa; Nagano, Kazuya; Arai, Masayoshi; Uyama, Hiroshi; Hirata, Yoshihiko; Muraoka, Misa

doi:https://doi.org/10.1007/s12010-024-05137-5

Effective Repeated Production of γ-glutamylcysteine, Essential For Intracellular Glutathione Production, Using Cellulose-immobilized Phytochelatin Synthase-like Enzyme NsPCS

Nakai, Takuya; Hirata, Kazumasa; Nagano, Kazuya et al.

Applied Biochemistry and Biotechnology, 2025

Number of Access:739（2025-08-28 23:13 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/100322

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Title	Effective Repeated Production of γ-glutamylcysteine, Essential For Intracellular Glutathione Production, Using Cellulose-immobilized Phytochelatin Synthase-like Enzyme NsPCS
Creator	CreatorName Nakai, Takuya
	NRID 1000030199062 1000030199062 CreatorName Hirata, Kazumasa
	NRID 1000040548301 1000040548301 CreatorName Nagano, Kazuya
	NRID 1000080311231 1000080311231 ORCID 0000-0003-2771-1948 0000-0003-2771-1948 researchmap permalink read0161265 read0161265 Scopus Author ID 7402957312 7402957312 CreatorName Arai, Masayoshi
	NRID 1000070203594 1000070203594 ORCID 0000-0002-8587-2507 0000-0002-8587-2507 researchmap permalink read0168389 read0168389 Scopus Author ID 7102108908 7102108908 CreatorName Uyama, Hiroshi
	CreatorName Hirata, Yoshihiko
	NRID 1000000707614 1000000707614 ORCID 0000-0001-8682-8906 0000-0001-8682-8906 researchmap permalink muraokamisa muraokamisa Scopus Author ID 55838089600 55838089600 CreatorName Muraoka, Misa
Description	The version of record of this article, first published in Applied Biochemistry and Biotechnology, is available online at Publisher’s website: https://doi.org/s12010-024-05137-5.
Abstract	γ-Glutamylcysteine (γ-EC) can increase intracellular glutathione (GSH) levels, which may prevent and alleviate age-related disorders and chronic diseases caused by oxidative damage. However, the commercial availability of γ-EC remains limited owing to its complex chemical synthesis from glutamate and cysteine. In this study, we have developed the method of the effective conversion of GSH to γ-EC to achieve the optimal reaction conditions for repeated batch production and potential application in industrial γ-EC production using the phytochelatin synthase-like enzyme NsPCS. For repeated batch conversion reactions, the optimal temperature was determined at 25 °C, where γ-EC showed good stability compared with that at 37 °C, leading to higher overall productivity. Cellulose sponges and microcrystalline cellulose (MCC) showed superior mechanical strength as immobilization carriers and greater stability and productivity than other materials. The total amounts of γ-EC obtained by NsPCS immobilized on the cellulose sponge and MCC were 305 mg and 291 mg, respectively, in a 5 mL reaction over five repeated batch reactions. These simple production processes are easily reproduced, and their high volumetric efficiency is promising for the industrial production of stable and low-cost γ-EC.
Publisher	Springer Science and Business Media LLC
Source Title	Applied Biochemistry and Biotechnology
Date of Issued	2025-01-21
Language	English
Handle URL	https://hdl.handle.net/11094/100322
PISSN	02732289
eISSN	15590291
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1007/s12010-024-05137-5
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
DCTERMS.bibliographicCitation	Applied Biochemistry and Biotechnology. 2025
DC.title	Effective Repeated Production of γ-glutamylcysteine, Essential For Intracellular Glutathione Production, Using Cellulose-immobilized Phytochelatin Synthase-like Enzyme NsPCS
DC.creator	Nakai, Takuya
	Hirata, Kazumasa
	Nagano, Kazuya
	Arai, Masayoshi
	Uyama, Hiroshi
	Hirata, Yoshihiko
	Muraoka, Misa
DC.publisher	Springer Science and Business Media LLC
DC.language' scheme='DCTERMS.RFC1766	English
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-01-21
DC.identifier	https://doi.org/10.1007/s12010-024-05137-5
DC.description	The version of record of this article, first published in Applied Biochemistry and Biotechnology, is available online at Publisher’s website: https://doi.org/s12010-024-05137-5.
DCTERMS.abstract	γ-Glutamylcysteine (γ-EC) can increase intracellular glutathione (GSH) levels, which may prevent and alleviate age-related disorders and chronic diseases caused by oxidative damage. However, the commercial availability of γ-EC remains limited owing to its complex chemical synthesis from glutamate and cysteine. In this study, we have developed the method of the effective conversion of GSH to γ-EC to achieve the optimal reaction conditions for repeated batch production and potential application in industrial γ-EC production using the phytochelatin synthase-like enzyme NsPCS. For repeated batch conversion reactions, the optimal temperature was determined at 25 °C, where γ-EC showed good stability compared with that at 37 °C, leading to higher overall productivity. Cellulose sponges and microcrystalline cellulose (MCC) showed superior mechanical strength as immobilization carriers and greater stability and productivity than other materials. The total amounts of γ-EC obtained by NsPCS immobilized on the cellulose sponge and MCC were 305 mg and 291 mg, respectively, in a 5 mL reaction over five repeated batch reactions. These simple production processes are easily reproduced, and their high volumetric efficiency is promising for the industrial production of stable and low-cost γ-EC.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	100322
citation_title	Effective Repeated Production of γ-glutamylcysteine, Essential For Intracellular Glutathione Production, Using Cellulose-immobilized Phytochelatin Synthase-like Enzyme NsPCS
citation_author	Nakai, Takuya
	Hirata, Kazumasa
	Nagano, Kazuya
	Arai, Masayoshi
	Uyama, Hiroshi
	Hirata, Yoshihiko
	Muraoka, Misa
citation_publisher	Springer Science and Business Media LLC
citation_language	English
citation_date	2025-01-21
citation_journal_title	Applied Biochemistry and Biotechnology
citation_doi	https://doi.org/10.1007/s12010-024-05137-5
citation_public_url	https://hdl.handle.net/11094/100322

Effective Repeated Production of γ-glutamylcysteine, Essential For Intracellular Glutathione Production, Using Cellulose-immobilized Phytochelatin Synthase-like Enzyme NsPCS

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