New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity
In this study, we established a new methodology for preparing 5′−nucleotidase (5′−NT) with the aim of enhancing our understanding of its enzyme activity and laying a basis for regulating the content of umami−enhancing nucleotides in pork. 5′−NT was prepared with Sephadex gel filtration and reverse−p...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/13/5/708 |
| _version_ | 1827319631718645760 |
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| author | Yin Zhang Qing Zeng Yingjie Zhang Pengcheng Zhang Qing Li Jiao Zhou Li Dong Zhongli Pan |
| author_facet | Yin Zhang Qing Zeng Yingjie Zhang Pengcheng Zhang Qing Li Jiao Zhou Li Dong Zhongli Pan |
| author_sort | Yin Zhang |
| collection | DOAJ |
| description | In this study, we established a new methodology for preparing 5′−nucleotidase (5′−NT) with the aim of enhancing our understanding of its enzyme activity and laying a basis for regulating the content of umami−enhancing nucleotides in pork. 5′−NT was prepared with Sephadex gel filtration and reverse−phase high−performance liquid chromatography, and its enzymatic properties and catalytic activity were evaluated. The results show that the molecular weight of the prepared 5′−NT was 57 kDa, the optimal catalytic temperature was 40 °C, and the optimal pH was 8. Zn<sup>2+</sup>, and sucrose showed inhibitory effects on the activity of 5′−NT, while K<sup>+</sup>, Na<sup>+</sup>, Ca<sup>2+</sup>, Mg<sup>2+</sup>, glucose, fructose, and trehalose promoted the activity of the studied compound. The prepared 5′−NT exhibited higher catalytic activity and selectivity against IMP compared with its commercial counterpart, while its catalytic activity against XMP was not significant (<i>p</i> > 0.05). In brief, we established a new methodology for preparing 5′−NT, enhancing our understanding of its enzyme activity and providing a solid basis for regulating the content of umami−enhancing nucleotides in pork through the control of endogenous 5′−NT activity. |
| first_indexed | 2024-04-25T00:29:48Z |
| format | Article |
| id | doaj.art-2c78a526bb6c49ef9eb853e13854bce8 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-04-25T00:29:48Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-2c78a526bb6c49ef9eb853e13854bce82024-03-12T16:44:11ZengMDPI AGFoods2304-81582024-02-0113570810.3390/foods13050708New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic ActivityYin Zhang0Qing Zeng1Yingjie Zhang2Pengcheng Zhang3Qing Li4Jiao Zhou5Li Dong6Zhongli Pan7Meat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaMeat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaMeat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaMeat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaMeat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaMeat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaMeat Processing Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610106, ChinaDepartment of Biological and Agricultural Engineering, University of California, Davis, One Shields Avenue, Davis, CA 95616, USAIn this study, we established a new methodology for preparing 5′−nucleotidase (5′−NT) with the aim of enhancing our understanding of its enzyme activity and laying a basis for regulating the content of umami−enhancing nucleotides in pork. 5′−NT was prepared with Sephadex gel filtration and reverse−phase high−performance liquid chromatography, and its enzymatic properties and catalytic activity were evaluated. The results show that the molecular weight of the prepared 5′−NT was 57 kDa, the optimal catalytic temperature was 40 °C, and the optimal pH was 8. Zn<sup>2+</sup>, and sucrose showed inhibitory effects on the activity of 5′−NT, while K<sup>+</sup>, Na<sup>+</sup>, Ca<sup>2+</sup>, Mg<sup>2+</sup>, glucose, fructose, and trehalose promoted the activity of the studied compound. The prepared 5′−NT exhibited higher catalytic activity and selectivity against IMP compared with its commercial counterpart, while its catalytic activity against XMP was not significant (<i>p</i> > 0.05). In brief, we established a new methodology for preparing 5′−NT, enhancing our understanding of its enzyme activity and providing a solid basis for regulating the content of umami−enhancing nucleotides in pork through the control of endogenous 5′−NT activity.https://www.mdpi.com/2304-8158/13/5/708porkumami5′−nucleotidaseisolation and purification |
| spellingShingle | Yin Zhang Qing Zeng Yingjie Zhang Pengcheng Zhang Qing Li Jiao Zhou Li Dong Zhongli Pan New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity Foods pork umami 5′−nucleotidase isolation and purification |
| title | New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity |
| title_full | New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity |
| title_fullStr | New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity |
| title_full_unstemmed | New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity |
| title_short | New Method for 5′−Nucleotidase Preparation and Evaluation of Its Catalytic Activity |
| title_sort | new method for 5 nucleotidase preparation and evaluation of its catalytic activity |
| topic | pork umami 5′−nucleotidase isolation and purification |
| url | https://www.mdpi.com/2304-8158/13/5/708 |
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