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Biochemical and Enzymatic Study of Rice BADH Wild-Type and Mutants: An Insight into Fragrance in Rice

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Abstract

Betaine aldehyde dehydrogenase 2 (BADH2) is believed to be involved in the accumulation of 2-acetyl-1-pyrroline (2AP), one of the major aromatic compounds in fragrant rice. The enzyme can oxidize ω-aminoaldehydes to the corresponding ω-amino acids. This study was carried out to investigate the function of wild-type BADHs and four BADH2 mutants: BADH2_Y420, containing a Y420 insertion similar to BADH2.8 in Myanmar fragrance rice, BADH2_C294A, BADH2_E260A and BADH2_N162A, consisting of a single catalytic-residue mutation. Our results showed that the BADH2_Y420 mutant exhibited less catalytic efficiency towards γ-aminobutyraldehyde but greater efficiency towards betaine aldehyde than wild-type. We hypothesized that this point mutation may account for the accumulation of γ-aminobutyraldehyde/Δ1-pyrroline prior to conversion to 2AP, generating fragrance in Myanmar rice. In addition, the three catalytic-residue mutants confirmed that residues C294, E260 and N162 were involved in the catalytic activity of BADH2 similar to those of other BADHs.

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Abbreviations

2AP:

2-Acetyl-1-pyrroline

ALDH:

Aldehyde dehydrogenase

AP-ald:

3-Aminopropionaldehyde

BADH:

Betaine aldehyde dehydrogenase

Bet-ald:

Betaine aldehyde

CD:

Circular dichroism

FPLC:

Fast protein liquid chromatography

GABA:

γ-Aminobutyric acid

GAB-ald:

γ-Aminobutyraldehyde

IPTG:

Isopropyl-β-d-thio-galactoside

SNPs:

Single nucleotide polymorphisms

TLC:

Thin layer chromatography

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Acknowledgments

This work is supported by grants from the Faculty of Science, Kasetsart University, the Commission on Higher Education, and the Agricultural Research Development Agency (public organization), Thailand.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand

    Ratree Wongpanya, Nonlawat Boonyalai, Napaporn Thammachuchourat & Kiattawee Choowongkomon

  2. Faculty of Medical Technology, Huachiew Chalermprakiet University, Samut Prakran, 10540, Thailand

    Natharinee Horata

  3. Rice Gene Discovery Unit, Kasetsart University, Kamphangsaen Campus, Nakhon Pathom, 73140, Thailand

    Siwaret Arikit, Khin Myo Myint & Apichart Vanavichit

  4. Department of Agronomy, International Program of Graduate School in Tropical Agriculture (RGJ Ph.D), Kamphaengsaen Campus, Nakon Pathom, 73140, Thailand

    Khin Myo Myint

  5. Center for Advanced Studies in Tropical Natural Resources, National Research University-Kasetsart University, Kasetsart University, Chatuchak, Bangkok, 10900, Thailand

    Kiattawee Choowongkomon

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  1. Ratree Wongpanya
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  2. Nonlawat Boonyalai
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Correspondence to Kiattawee Choowongkomon.

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Wongpanya, R., Boonyalai, N., Thammachuchourat, N. et al. Biochemical and Enzymatic Study of Rice BADH Wild-Type and Mutants: An Insight into Fragrance in Rice. Protein J 30, 529–538 (2011). https://doi.org/10.1007/s10930-011-9358-5

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  • DOI: https://doi.org/10.1007/s10930-011-9358-5

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