Construction of cell factories for production of valencene in Saccharomyces cerevisiae / 药学学报

Valencene, a kind of sesquiterpenoid with a citrus flavor, is mainly found in Valencia orange and is commonly used in cosmetics and food additives, as well as industrial synthetic nootkatone. In this study, synthetic biology was used to create a Saccharomyces cerevisiae cell factory to produce valencene. Fistly, valencene synthase gene (CnVS) from Callitropsis nootkatensis was inserted into the chromosome of the chassis strain YTT-T5. The resulting strain VAL-01 could produce 1.1 mg·L-1 valencene. Protein fusion technique was used, different valencene synthases were compared and the copy number of key genes was adjusted, yielding valencene to 436.4 mg·L-1. Then, knocking-out the transcription factor ROX1 resulted in valencene improvement by 17.4%. Moreover, the induction system of galactose was regulated, transcription factor PDR3 and INO2 were overexpressed. The engineered strain VAL-10 could produce 2 798.6 mg·L-1 valencene by high cell density fermentation method (nearly 2 500 times higher than VAL-01). This study provides a basis for green production of valencene.

Valencene-rich fraction from Vetiveria zizanioides exerts immunostimulatory effects in vitro and in mice

Objective: To decipher the responsible compound present in the aqueous root extract of Vetiveria zizanioides which has tremendous immunomodulatory activity. Methods: Different fractions of the water extract were collected and analyzed for immunomodulatory activity by analyzing in vitro phagocytic activity and nitric oxide production. One fraction VF3 was selected and further analyzed for possible compounds by high performance liquid chromatography and gas chromatography coupled with a mass spectrometer. The in vitro immunomodulatory parameters such as phagocytic index, nitrite content, and tumor necrosis factor-α production in murine macrophages were analyzed. In vivo studies, sheep red blood cell induced haemagglutination titer, the number of antibody-producing cells, and sheep red blood cell induced delayed-type hypersensitivity were analyzed. Cytotoxic studies in L929 normal fibroblasts were also performed. Results: One of the fractions, VF3, was selected and confirmed the presence of an active compound valencene. The in vitro immunomodulatory parameters were significantly (P<0.05) increased by valencene treatment. In vivo studies in Swiss albino mice showed that valencene could significantly (P<0.05) increase haemagglutination titer, the number of antibody-producing cells, and delayed-type hypersensitivity. Cytotoxic studies also showed that valencene did not cause any morphological changes and DNA damage in normal fibroblasts. Conclusions: Valencene possesses immunomodulatory activities and can be commercially exploited for its immunostimulatory potentials.