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Computational design and selections for an engineered, thermostable terpene synthase.

ABSTRACT: Terpenoids include structurally diverse antibiotics, flavorings, and fragrances. Engineering terpene synthases for control over the synthesis of such compounds represents a long sought goal. We report computational design, selections, and assays of a thermostable mutant of tobacco 5-epi-aristolochene synthase (TEAS) for the catalysis of carbocation cyclization reactions at elevated temperatures. Selection for thermostability included proteolytic digestion followed by capture of intact proteins. Unlike the wild-type enzyme, the mutant TEAS retains enzymatic activity at 65°C. The thermostable terpene synthase variant denatures above 80°C, approximately twice the temperature of the wild-type enzyme.

SUBMITTER: Diaz JE

PROVIDER: S-EPMC3190154 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature

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Computational design and selections for an engineered, thermostable terpene synthase.

Diaz Juan E JE Lin Chun-Shi CS Kunishiro Kazuyoshi K Feld Birte K BK Avrantinis Sara K SK Bronson Jonathan J Greaves John J Saven Jeffery G JG Weiss Gregory A GA

Protein science : a publication of the Protein Society 20110802 9

Terpenoids include structurally diverse antibiotics, flavorings, and fragrances. Engineering terpene synthases for control over the synthesis of such compounds represents a long sought goal. We report computational design, selections, and assays of a thermostable mutant of tobacco 5-epi-aristolochene synthase (TEAS) for the catalysis of carbocation cyclization reactions at elevated temperatures. Selection for thermostability included proteolytic digestion followed by capture of intact proteins. ...[more]

PMID: 21739507

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