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. 2018 Dec 18;8:17936. doi: 10.1038/s41598-018-36302-6

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Biochemical characterization of PtTPS16 mutants generated using in vitro mutagenesis. GC-MS chromatograms representing the product spectra of wild type PtTPS16 (A), wild type PtTPS21 (L), and the different PtTPS16 mutants (B–K) are shown. Amino acid changes (one letter code) and their positions relative to the PtTPS16 sequence are indicated in the name of the mutants. The genes were heterologously expressed in E. coli and partially purified proteins were incubated with GPP as substrate. Enzyme products were analyzed using GC-MS. 1, α-thujene; 2, α-pinene*; 3, camphene*; 4, sabinene; 5, β-pinene*; 6, myrcene; 7, α-phellandrene; 8, α-terpinene*; 9, β-phellandrene; 10, limonene*; 11, ocimene; 12, γ-terpinene*; 13, terpinolene. Compounds marked with * were identified by comparison of retention time and mass spectrum to those of authentic standards. Others were identified by database comparisons.