Abstract
The genetic improvement of enzymes important in the brewing process is one of the main goals of barley biotechnology. For the improvement of β-amylase thermostability in barley seeds, we have already constructed a mutant thermostable β-amylase gene, using site-directed mutagenesis and random mutagenesis to achieve the substitution of seven amino acids of the original barley β-amylase. This sevenfold-mutant barley β-amylase showed a thermostability increased by 11.6 °C compared to the original enzyme. In the present article, a thermostable β-amylase gene under the control of the barley β-amylase promoter was introduced to barley protoplasts, and fertile plants were generated from 9 independent transgenic lines. Subsequent analyses indicated that the thermostable β-amylase gene was expressed and β-amylase activity derived from both native and modified genes was detected in the seeds of 6 transgenic lines. The transgene was stably transmitted to progeny, and thermostable β-amylase was synthesized in T4 seeds, demonstrating that our strategy is applicable for the improvement of seed quality for industrial utilization.
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