Development and Identification of Thermostable Tryptophanase Based on Ancestral Sequence Analysis
Yulei Zhang1,2, Yun Li1, Heng Hu1
1Anhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Metabolic Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui 241000, People's Republic of China.
Abstract:
Indole and its derivatives are valuable compounds used in flavor, fragrance, and pharmaceutical industries. Tryptophanase (TnaA, EC 4.1.99.1) catalyzes the conversion of L-tryptophan into indole, pyruvate, and ammonia, but limited thermostability restricts its industrial application. Here, a thermostable TnaA from Morganella morganii was identified through ancestral sequence analysis, structural prediction, and molecular dynamics simulations. The enzyme existed mainly as a dimer under native conditions and showed optimal activity at 55 °C and pH 8.0. Compared with the commonly used Proteus vulgaris TnaA, M. morganii TnaA exhibited an 8.8 °C higher Tm, a 9.1 °C higher T5030, and an 18-fold longer half-life at 45 °C. Substrate specificity analysis showed activity only toward L-tryptophan and l-serine. These properties make M. morganii TnaA a promising biocatalyst for thermostable indole production.
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