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Amino acid substitutions of the limit dextrinase gene in barley are associated with enzyme thermostability

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Abstract

Limit dextrinase (LD) is a key enzyme in determining the malting quality. A survey of 60 barley varieties showed a wide range of variation for the enzyme activity and thermostability. Galleon showed low enzyme activity and high thermostability while Maud showed high activity and low thermostability. Alignment of the LD amino acid sequences of Galleon and Maud identified seven amino acid substitutions Lys/Arg-102, Thr/Ala-233, Ser/Gly-235, Gly/Ala-298, Cys/Arg-415, Ala/Ser-885 and Gly/Cys-888. Genetic diversity of LD was investigated using single strand conformation polymorphism based on the amino acid substitutions. Only limited genetic variation was detected in the current malting barley varieties, although wide variation was observed in the wider barley germplasm. The Thr/Ala-233 and Ala/Ser-885 substitutions were associated with enzyme thermostability (P < 0.0001), but no polymorphism was associated with the enzyme activity. This result was confirmed from further sequence analysis. The results will provide a tool for understanding and selection of high LD thermostability.

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Acknowledgements

This research is supported by the Australian Grain Research & Development Corporation project UT00012 and Australia-China Collaborative Project.

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Authors and Affiliations

  1. National Natural Science Foundation of China, 83 Shuangqing Road, Haidian District, Beijing, 100085, China

    Xinquan Yang

  2. Department of Agriculture and Food, Agricultural Research Western Australia, South Perth, WA, 6151, Australia

    Xinquan Yang, Sharon Westcott, Xue Gong, Reg C. M. Lance & Chengdao Li

  3. Tasmanian Institute of Agricultural Research, University of Tasmania, Private Bag 54, Hobart, TAS, 7001, Australia

    Evan Evans

  4. Western Australian State Agricultural Biotechnology Centre, Murdoch University, Murdoch, WA, 6051, Australia

    Xiao-Qi Zhang, Reg C. M. Lance & Chengdao Li

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  1. Xinquan Yang
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  2. Sharon Westcott
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  3. Xue Gong
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  4. Evan Evans
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  5. Xiao-Qi Zhang
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  7. Chengdao Li
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Correspondence to Chengdao Li.

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Yang, X., Westcott, S., Gong, X. et al. Amino acid substitutions of the limit dextrinase gene in barley are associated with enzyme thermostability. Mol Breeding 23, 61–74 (2009). https://doi.org/10.1007/s11032-008-9214-2

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  • DOI: https://doi.org/10.1007/s11032-008-9214-2

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