Updated on 2025/05/02

写真a

 
潤井 みや
 
Organization
Faculty of Medicine, Dentistry and Pharmaceutical Sciences Special-Appointment Assistant Professor
Position
Special-Appointment Assistant Professor
External link

Degree

  • PhD ( 2024.3   Kobe Pharmaceutical University )

Research Interests

  • Alkaloids

  • Transporter

  • 植物特化代謝産物

  • 合成生物学

  • Komagataella phaffii

  • Pichia pastoris

  • 脂質

  • 薬用植物

Education

  • 大学女性協会 国内奨学生    

    2022.4 - 2024.3

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  • 日本薬学会 長井記念薬学研究奨励金奨学生    

    2022.4 - 2024.3

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  • Kobe Pharmaceutical University   薬学研究科   博士課程 (4年)

    2020.4 - 2024.3

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  • Kobe Pharmaceutical University   薬学部  

    2014.4 - 2020.3

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Research History

  • Okayama University   Graduate School of Medicine , Dentistry and Pharmaceutical Sciences

    2025.4

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  • Shiga University of Medical Science

    2024.4 - 2025.2

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Papers

  • Integration of co-culture and transport engineering for enhanced metabolite production

    Yasuyuki Yamada, Miya Urui, Nobukazu Shitan

    Plant Biotechnology   41 ( 3 )   195 - 202   2024.9

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    Publishing type:Research paper (scientific journal)   Publisher:Japanese Society for Plant Cell and Molecular Biology  

    DOI: 10.5511/plantbiotechnology.24.0312b

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  • Enhanced Co-culture System Using Escherichia coli and Pichia pastoris (Komagataella phaffii) for Improved Microbial Production of Valuable Plant Alkaloids.

    Miya Urui, Yasuyuki Yamada, Akira Nakagawa, Fumihiko Sato, Hiromichi Minami, Nobukazu Shitan

    Biological & pharmaceutical bulletin   46 ( 10 )   1494 - 1497   2023

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    Language:English   Publishing type:Research paper (scientific journal)  

    Advancements in synthetic biology have facilitated the microbial production of valuable plant metabolites. However, constructing complete biosynthetic pathways within a single host organism remains challenging. To solve this problem, modular co-culture systems involving host organisms with partial pathways have been developed. We focused on Escherichia coli, a general host for metabolite production, and Pichia pastoris (Komagataella phaffii), a novel synthetic biology host due to its high expression of biosynthetic enzymes. Previously, we reported the co-culture of E. coli cells, which produce reticuline (an important intermediate for various alkaloids) from glycerol, with P. pastoris cells, which produce the valuable alkaloid stylopine from reticuline. However, Pichia cells inhibited E. coli growth and reticuline production. Therefore, we aimed to improve this co-culture system. We investigated the pre-culture time before co-culture to enhance E. coli growth and reticuline production. Additionally, we examined the optimal concentration of Pichia cells inoculated for co-culture and methanol addition during co-culture for the continuous expression of biosynthetic enzymes in Pichia cells. We successfully established an improved co-culture system that exhibited an 80-fold increase in productivity compared to previous methods. This enhanced system holds great potential for the rapid and large-scale production of various valuable plant metabolites.

    DOI: 10.1248/bpb.b23-00473

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  • Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli. International journal

    Yasuyuki Yamada, Miya Urui, Hidehiro Oki, Kai Inoue, Haruyuki Matsui, Yoshito Ikeda, Akira Nakagawa, Fumihiko Sato, Hiromichi Minami, Nobukazu Shitan

    Metabolic engineering communications   13   e00184   2021.12

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    Language:English   Publishing type:Research paper (scientific journal)  

    Microorganisms can be metabolically engineered to produce specialized plant metabolites. However, these methods are limited by low productivity and intracellular accumulation of metabolites. We sought to use transport engineering for producing reticuline, an important intermediate in the alkaloid biosynthetic pathway. In this study, we established a reticuline-producing Escherichia coli strain into which the multidrug and toxic compound extrusion transporter Arabidopsis AtDTX1 was introduced. AtDTX1 was selected due to its suitable expression in E. coli and its reticuline-transport activity. Expression of AtDTX1 enhanced reticuline production by 11-fold, and the produced reticuline was secreted into the medium. AtDTX1 expression also conferred high plasmid stability and resulted in upregulation or downregulation of several genes associated with biological processes, including metabolic pathways for reticuline biosynthesis, leading to the production and secretion of high levels of reticuline. The successful employment of a transporter for alkaloid production suggests that the proposed transport engineering approach may improve the biosynthesis of specialized metabolites via metabolic engineering.

    DOI: 10.1016/j.mec.2021.e00184

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  • Establishment of a co-culture system using Escherichia coli and Pichia pastoris (Komagataella phaffii) for valuable alkaloid production. International journal

    Miya Urui, Yasuyuki Yamada, Yoshito Ikeda, Akira Nakagawa, Fumihiko Sato, Hiromichi Minami, Nobukazu Shitan

    Microbial cell factories   20 ( 1 )   200 - 200   2021.10

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    Language:English   Publishing type:Research paper (scientific journal)  

    BACKGROUND: Plants produce a variety of specialized metabolites, many of which are used in pharmaceutical industries as raw materials. However, certain metabolites may be produced at markedly low concentrations in plants. This problem has been overcome through metabolic engineering in recent years, and the production of valuable plant compounds using microorganisms such as Escherichia coli or yeast cells has been realized. However, the development of complicated pathways in a single cell remains challenging. Additionally, microbial cells may experience toxicity from the bioactive compounds produced or negative feedback effects exerted on their biosynthetic enzymes. Thus, co-culture systems, such as those of E. coli-E. coli and E. coli-Saccharomyces cerevisiae, have been developed, and increased production of certain compounds has been achieved. Recently, a co-culture system of Pichia pastoris (Komagataella phaffii) has gained considerable attention due to its potential utility in increased production of valuable compounds. However, its co-culture with other organisms such as E. coli, which produce important intermediates at high concentrations, has not been reported. RESULTS: Here, we present a novel co-culture platform for E. coli and P. pastoris. Upstream E. coli cells produced reticuline from a simple carbon source, and the downstream P. pastoris cells produced stylopine from reticuline. We investigated the effect of four media commonly used for growth and production of P. pastoris, and found that buffered methanol-complex medium (BMMY) was suitable for P. pastoris cells. Reticuline-producing E. coli cells also showed better growth and reticuline production in BMMY medium than that in LB medium. De novo production of the final product, stylopine from a simple carbon source, glycerol, was successful upon co-culture of both strains in BMMY medium. Further analysis of the initial inoculation ratio showed that a higher ratio of E. coli cells compared to P. pastoris cells led to higher production of stylopine. CONCLUSIONS: This is the first report of co-culture system established with engineered E. coli and P. pastoris for the de novo production of valuable compounds. The co-culture system established herein would be useful for increased production of heterologous biosynthesis of complex specialized plant metabolites.

    DOI: 10.1186/s12934-021-01687-z

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MISC

Presentations

  • 胆汁酸肝組織障害の改善を目指したリン脂質トランスポーターABCB4の機能解明

    池田義人, 小澤慶祐, 潤井みや, 山田泰之, 士反伸和, 森田真也

    膜シンポジウム2024  2024.11.29 

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    Event date: 2024.11.28 - 2024.11.29

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  • 植物有用成分など医薬品原料の安定供給を指向した大腸菌-ピキア酵母共培養系の確立と輸送工学 Invited

    潤井みや

    医療薬学フォーラム2024/第32回クリニカルファーマーシーシンポジウム  2024.7.6 

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    Event date: 2024.7.6 - 2024.7.7

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  • 植物有用成分など医薬品原料の安定供給を指向した合成生物学における共培養系の応用

    潤井 みや, 山田 泰之, 池田義人, 中川明, 佐藤 文彦, 南 博道, 士反 伸和

    第17回次世代を担う若手のための医療薬科学シンポジウム  2023.9.17 

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    Presentation type:Oral presentation (general)  

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  • 有用アルカロイド高生産を目指した大腸菌−ピキア酵母共培養系の応用

    潤井 みや, 山田 泰之, 池田義人, 中川明, 佐藤 文彦, 南 博道, 士反 伸和

    日本薬学会143年会  2023.3.27 

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  • 大腸菌とピキア酵母の共培養による効率的なアルカロイド生産系の開発

    潤井 みや, 山田 泰之, 池田義人, 中川明, 佐藤 文彦, 南 博道, 士反 伸和

    日本生薬学会第68回年会  2022.9.10 

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  • アルカロイド生産性大腸菌とピキア酵母の共培養系の確立と増産への応用

    潤井 みや, 山田 泰之, 池田義人, 中川明, 佐藤 文彦, 南 博道, 士反 伸和

    日本薬学会 第142年会  2022.3.25 

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  • 有用物質生産を目指した微生物共培養系の確立と輸送工学を用いた増産への応用

    潤井 みや, 山田 泰之, 池田義人, 中川明, 佐藤 文彦, 南 博道, 士反 伸和

    2021.11.19 

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    Presentation type:Poster presentation  

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  • Establishment of a Co-Culture System Using Escherichia coli and Pichia pastoris to Produce Valuable Alkaloids

    2021.9.15 

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  • 大腸菌とピキア酵母の共培養による効率的なスチロピンアルカロイドの生産

    潤井 みや, 山田 泰之, 池田義人, 南 博道, 佐藤 文彦, 士反 伸和

    第38回日本植物バイオテクノロジー学会(つくば)大会  2021.9.9 

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  • 大腸菌とピキア酵母の共培養系の開発ー植物有用アルカロイド生産を目指してー

    潤井 みや, 山田 泰之, 池田義人, 南 博道, 佐藤 文彦, 士反 伸和

    生合成若手シンポジウム2021  2021.8.7 

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    Presentation type:Poster presentation  

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  • Establishment of a Co-Culture System Using Escherichia coli and Pichia Pastoris for Valuable Alkaloid Production

    2021.7.11 

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  • ピキア酵母用培地によるアルカロイド生産性ピキア酵母および大腸菌のアルカロイド生産性の検討

    潤井 みや, 山田 泰之, 池田義人, 南 博道, 佐藤 文彦, 士反 伸和

    日本薬学会141年会  2021.3.26 

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  • アルカロイド生産性ピキア酵母を用いた培地によるアルカロイド生産性の検討

    潤井 みや, 山田 泰之, 南 博道, 佐藤 文彦, 士反 伸和

    第70回日本薬学会関西支部  2020.10.10 

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  • Expression of an alkaloid transporter NtJAT1 enhances the alkaloid production rate in stylopine-producing Pichia cells

    2020.3.25 

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  • アルカロイド生産性ピキア酵母への輸送体タンパク質の導入と生産性変化の検討

    潤井 みや, 山田 泰之, 大木 秀浩, 炭田 奈々, 南 博道, 佐藤 文彦, 士反 伸和

    第69回日本薬学会関西支部  2019.10.12 

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  • Stylopine生産性ピキア酵母への耐性・輸送体タンパク質の導入と生産性変化の検討

    潤井 みや, 山田 泰之, 大木 秀浩, 炭田 奈々, 南 博道, 佐藤 文彦, 士反 伸和

    第14回トランスポーター研究会  2019.7.20 

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Awards

  • 優秀発表賞

    2023.9   第17回次世代を担う若手のための医療薬科学シンポジウム   植物有用成分など医薬品原料の安定供給を指向した合成生物学における共培養系の応用

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  • 学生優秀発表賞

    2023.3   日本薬学会143年会   有用アルカロイド高生産を目指した大腸菌−ピキア酵母共培養系の応用

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  • 優秀口頭発表賞

    2020.10   第70回日本薬学会関西支部   アルカロイド生産性ピキア酵母を用いた培地によるアルカロイド生産性の検討

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