Limonene and its derivative perillic acid are widely used in food, cosmetics, health products, medicine and other industries as important bioactive natural products. However, inefficient plant extraction and high energy-consuming chemical synthesis hamper the industrial production of limonene and perillic acid. In this study, limonene synthase from Mentha spicata was expressed in Saccharomyces cerevisiae by peroxisome compartmentalization, and the yield of limonene was 0.038 mg/L. The genes involved in limonene synthesis, ERG10, ERG13, tHMGR, ERG12, ERG8, IDI1, MVD1, ERG20ww and tLS, were step-wise expressed via modular engineering to study their effects on limonene yield. The yield of limonene increased to 1.14 mg/L by increasing the precursor module. Using the plasmid with high copy number to express the above key genes, the yield of limonene significantly increased up to 86.74 mg/L, which was 4 337 times higher than that of the original strain. Using the limonene-producing strain as the starting strain, the production of perillic acid was successfully achieved by expressing cytochrome P450 enzyme gene from Salvia miltiorrhiza, and the yield reached 4.42 mg/L. The results may facilitate the construction of cell factory with high yield of monoterpene products by S. cerevisiae.
Saccharomyces cerevisiae/metabolism* ; Limonene/metabolism* ; Metabolic Engineering ; Monoterpenes/metabolism*

This study aims to explore the consistency between macroscopic identification and DNA barcoding identification of Amomi Fructus. With the DNA barcoding identification results, we evaluated the reliability of identifying Amomi Fructus quality by combining macroscopic traits with main volatile chemical components. Thirteen batches of Amomi Fructus samples were collected for identification. Firstly, the morphological and sensory characteristics of each sample were observed and recorded according to the standard in Chinese Pharmacopoeia(2020 edition). The 100-fruit weight, longitudinal diameter, transverse diameter, and longitudinal diameter-to-transverse diameter ratio were measured, which correspond to large, solid, and full kernel representing good quality in the sensory evaluation. The odor value detected by electronic nose and major volatile components(borneol, camphor, limonene, and borneol acetate) correspond to the sensory evaluation of strong odor representing good quality. Secondly, DNA barcoding was employed to identify the 13 batches of samples. Finally, clustering analysis was performed for the main volatile components and macroscopic traits, and the identification results were compared with those of DNA barcoding. Except two batches of samples(No.6 and No.10), the macroscopic identification showed the results consistent with those of DNA barcoding, with an identification rate of 84.62%. The clustering results of the content of four volatile chemical components and macroscopic traits were also consistent with the DNA barcoding identification results. DNA barcoding can verify the results of macroscopic identification and provide a scientific basis for the inheritance and development of macroscopic identification. Moreover, the combination of macroscopic traits and chemical components demonstrates higher accuracy in the quality evaluation of Chinese medicinal materials.
Camphanes ; Camphor/analysis* ; DNA Barcoding, Taxonomic ; Drugs, Chinese Herbal/chemistry* ; Fruit/genetics* ; Limonene/analysis* ; Reproducibility of Results

Since Curcumae Radix decoction pieces have multiple sources, it is difficult to distinguish depending on traditional cha-racters, and the mixed use of multi-source Curcumae Radix will affect its clinical efficacy. Heracles Neo ultra-fast gas phase electronic nose was used in this study to quickly identify and analyze the odor components of 40 batches of Curcumae Radix samples from Sichuan, Zhejiang, and Guangxi. Based on the odor fingerprints established for Curcumae Radix decoction pieces of multiple sources, the odor components was identified and analyzed, and the chromatographic peaks were processed and analyzed to establish a rapid identification method. Principal component analysis(PCA), discriminant factor analysis(DFA), and soft independent modeling cluster analysis(SIMCA) were constructed for verification. At the same time, one-way analysis of variance(ANOVA) combined with variable importance in projection(VIP) was employed to screen out the odor components with P<0.05 and VIP>1, and 13 odor components such as β-caryophyllene and limonene were hypothesized as the odor differential markers of Curcumae Radix decoction pieces of diffe-rent sources. The results showed that Heracles Neo ultra-fast gas phase electronic nose can well analyze the odor characteristics and rapidly and accurately discriminate Curcumae Radix decoction pieces of different sources. It can be applied to the quality control(e.g., online detection) in the production of Curcumae Radix decoction pieces. This study provides a new method and idea for the rapid identification and quality control of Curcumae Radix decoction pieces.
Drugs, Chinese Herbal/analysis* ; Electronic Nose ; China ; Plant Roots/chemistry* ; Limonene/analysis* ; Chromatography, High Pressure Liquid

OBJECTIVE: To explore the effect of mucoregulatory agents during endoscopic sinus surgery. METHOD: Ninety-seven cases with chronic rhinosinusitis were randomly divided into three groups, with 31 cases in B group treated by ambroxol, 33 cases in C group treated by eucalyptol-limonene-pinene enteric soft capsule and 33 cases in control group (A group). The follow-up visit lasted for 6 months for three groups. Then, the therapeutic effects were evaluated and compared among these three groups. RESULT: By the end of 6 months after treatment,the effective rate was 90.3% and 97.0% for cases in the B and C groups respectively, and only 75.8% in A group. The difference between A and C was statistically significant (P < 0.05). CONCLUSION Eucalyptol-limonene-pinene enteric soft capsule,as a multicomponent mucoregulatory agent, can obviously improve the secretion of mucosa and epithelial recovery, thus accelerate healing of the disease. It can also improve the success rate of functional endoscope sinus surgery, and may play a promising role in clinical application.
Adolescent ; Adult ; Aged ; Ambroxol ; therapeutic use ; Chronic Disease ; Cyclohexanols ; therapeutic use ; Cyclohexenes ; therapeutic use ; Eucalyptol ; Expectorants ; therapeutic use ; Female ; Humans ; Intraoperative Period ; Limonene ; Male ; Middle Aged ; Monoterpenes ; therapeutic use ; Sinusitis ; drug therapy ; Terpenes ; therapeutic use ; Treatment Outcome ; Young Adult