Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Dec 20;13(1):17.
doi: 10.3390/foods13010017.

The Optimization of Assay Conditions and Characterization of the Succinic Semialdehyde Dehydrogenase Enzyme of Germinated Tartary Buckwheat

Yuchan Yang  1 , Jiashang Liu  2 , Nan Li  1 , Yu Guo  1 , Hua Ye  1 , Zhanming Li  1 , Dongxu Wang  1 , Yuanxin Guo  1
Affiliations

The Optimization of Assay Conditions and Characterization of the Succinic Semialdehyde Dehydrogenase Enzyme of Germinated Tartary Buckwheat

Yuchan Yang et al. Foods. .

Abstract

In this study, the conditions for optimizing the determination of succinic semialdehyde dehydrogenase (SSADH, EC 1.2.1.79) activity in germinated Tartary buckwheat were investigated. Based on a single-factor test, the effects of temperature, pH, and succinic semialdehyde (SSA) concentration on the enzyme activity of germinated buckwheat SSADH were investigated by using the response surface method, and optimal conditions were used to study the enzymatic properties of germinated buckwheat SSADH. The results revealed that the optimum conditions for determining SSADH enzyme activity are as follows: temperature-30.8 °C, pH-8.7, and SSA concentration-0.3 mmol/L. Under these conditions, SSADH enzyme activity was measured as 346 ± 9.61 nmol/min. Furthermore, the thermal stability of SSADH was found to be superior at 25 °C, and its pH stability remained comparable at pH levels of 7.6, 8.1, and 8.6 in germinated Tartary buckwheat samples; however, a decline in stability was observed at pH 9.1. Cu2+, Co2+, and Ni2+ exhibited an activating effect on SSADH activity in germinating Tartary buckwheat, with Cu2+ having the greatest influence (p < 0.05), which was 1.21 times higher than that of the control group. Zn2+, Mn2+, and Na+ inhibited SSADH activity in germinating Tartary buckwheat, with Zn2+ showing the strongest inhibitory effect (p < 0.05). On the other hand, the Km and Vmax of SSADH for SSA in germinated Tartary buckwheat were 0.24 mmol/L and 583.24 nmol/min. The Km and Vmax of SSADH for NAD+ in germinated Tartary buckwheat were 0.64 mmol/L and 454.55 nmol/min.

Keywords: enzymatic characteristics; enzyme activity assay; germinated Tartary buckwheat; response surface analysis; succinic semialdehyde dehydrogenase.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Graded sedimentation of (NH4)2SO4 from the SSADH of germinated Tartary buckwheat.
Figure 2
Figure 2
Effect of temperature on the SSADH activity of germinated Tartary buckwheat.
Figure 3
Figure 3
Effects of different pH values on the SSADH activity of germinated Tartary buckwheat.
Figure 4
Figure 4
Effect of SSA concentration on SSADH determination of enzyme activity of germinated Tartary buckwheat.
Figure 5
Figure 5
Correlation between predicted and experimental results regarding SSADH activity content.
Figure 6
Figure 6
Response surface and contour map of the interaction between temperature and pH on SSADH activity of germinated Tartary buckwheat (SSA 0.2 mmol/L).
Figure 7
Figure 7
Response surface and contour map of the interaction between temperature and SSA concentration on SSADH enzyme activity of germinated Tartary buckwheat (pH 8.6).
Figure 8
Figure 8
Response surface and contour map of the interaction between pH and SSA concentration on SSADH activity of germinated Tartary buckwheat (30 °C).
Figure 9
Figure 9
Thermal stability of germinated Tartary buckwheat.
Figure 10
Figure 10
pH stability of germinated Tartary buckwheat.
Figure 11
Figure 11
Effects of different metal ions on the SSADH activity of germinated Tartary buckwheat Different Superscript letters (a, b, c, d, e) indicate statistically significant differences among groups.
Figure 12
Figure 12
Effect of SSA concentration on the SSADH activity of germinated Tartary buckwheat.
Figure 13
Figure 13
Lineweaver–Burk dynamics chart of the SSADH of germinated Tartary buckwheat (SSA).
Figure 14
Figure 14
Effect of coenzyme NAD+ concentration on the SSADH activity of germinated Tartary buckwheat.
Figure 15
Figure 15
Lineweaver–Burk dynamics chart of SSADH of germinated Tartary buckwheat (NAD+).
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Wu D.-T., Wang J., Li J., Hu J.-L., Yan H., Zhao J., Zou L., Hu Y.-C. Physicochemical properties and biological functions of soluble dietary fibers isolated from common and Tartary buckwheat sprouts. LWT. 2023;183:114944. doi: 10.1016/j.lwt.2023.114944. - DOI
    1. Liu M., Ma Z., Zheng T., Sun W., Zhang Y., Jin W., Zhan J., Cai Y., Tang Y., Wu Q., et al. Insights into the correlation between Physiological changes in and seed development of tartary buckwheat (Fagopyrum tataricum Gaertn.) BMC Genom. 2018;19:648. doi: 10.1186/s12864-018-5036-8. - DOI - PMC - PubMed
    1. Li D., Li X., Ding X. Composition and antioxidative properties of the flavonoid-rich fractions from tartary buckwheat grains. Food Sci. Biotechnol. 2010;19:711–716. doi: 10.1007/s10068-010-0100-4. - DOI
    1. Zhou X., Hao T., Zhou Y., Tang W., Xiao Y., Meng X., Fang X. Relationships between antioxidant compounds and antioxidant activities of tartary buckwheat during germination. J. Food Sci. Technol. 2015;52:2458–2463. doi: 10.1007/s13197-014-1290-1. - DOI - PMC - PubMed
    1. Bao T., Wang Y., Li Y., Gowd V., Niu X., Yang H., Chen L., Chen W., Sun C. Antioxidant and antidiabetic properties of tartary buckwheat rice flavonoids after in vitro digestion. J. Zhejiang Univ. Sci. B. 2016;17:941–951. doi: 10.1631/jzus.B1600243. - DOI - PMC - PubMed