Isolation, purification, and mass spectrometry identification of the enzyme involved in citrus flavor (+)-valencene biotransformation to (+)-nootkatone by Yarrowia lipolytica.

BACKGROUND: (+)-Nootkatone is a highly valuable sesquiterpene compound that can be used as an aromatic in the food industry because of its grapefruit flavor and low sensory threshold. The unconventional yeast Yarrowia lipolytica has many unique physical and chemical properties, metabolic characteristics, and genetic structure, which has aroused the interest of researchers. Previous research showed that Y. lipolytica possesses the ability to transform the sesquiterpene (+)-valencene to (+)-nootkatone. The aim of this study was to isolate, purify, and identify the enzyme involved in the (+)-valencene bioconversion to (+)-nootkatone by Y. lipolytica. RESULTS: In this study, ultrasonic-assisted extraction, ammonium sulfate precipitation, anion-exchange chromatography, and gel-filtration chromatography were used to separate and purify the enzyme involved in the (+)-valencene bioconversion by Y. lipolytica. The protein was identified as aldehyde dehydrogenase (ALDH) (gene0658) using sodium dodecyl sulfate polyacrylamide gel electrophoresis and liquid chromatography-tandem mass spectrometry analysis. The ALDH had the highest activity when the pH value was 6.0 and the temperature was 30 °C. The activity of ALDH was significantly stimulated by ferrous ions and inhibited by barium, calcium, and magnesium ions. CONCLUSION: This is the first time that ALDH was found to participate in (+)-valencene biotransformation by Y. lipolytica. It may be involved in regulating the microbial transformation of (+)-valencene to (+)-nootkatone through redox characteristics. This study provides a theoretical basis and reference for the biological synthesis of citrus flavor (+)-nootkatone. © 2023 Society of Chemical Industry.

Protective effect of orange essential oil on the formation of non-alcoholic fatty liver disease caused by high-fat diet.

The purpose of this study was to investigate the protective effect of sniffing orange essential oil (OEO) on the formation of non-alcoholic fatty liver disease (NAFLD) caused by a high-fat diet. The results confirmed that sniffing OEO could reduce obesity caused by a high-fat diet (HFD) by reducing the levels of triglycerides (TGs), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C). In addition, the observation of liver tissue sections showed that sniffing OEO could reduce lipid accumulation in liver cells. Further analysis by western blot analysis showed that OEO treatment made the expression levels of acetyl-CoA carboxylase (ACC) and Cytochrome P450 2E1 (CYP2E1) down-regulated and the expression levels of peroxisome proliferator-activated receptor-α (PPAR-α) and carnitine palmitoyltransferase-1 (CPT-1) up-regulated. These results indicate that the treatment of sniffing OEO could enhance the antioxidant capacity of mice and reduce liver damage caused by a high-fat diet. Furthermore, sniffing OEO could inhibit lipid synthesis and oxidative stress stimulated by a high-fat diet. Overall, OEO treatment had a certain protective effect on NAFLD-related diseases caused by a high-fat diet. Therefore, aromatherapy may be introduced as a treatment of long-term chronic diseases.

Extraction of orange pectin based on the interaction between sodium caseinate and pectin.

The interaction between commercial orange pectin (COP) and sodium caseinate (SC) was studied using FTIR, fluorescence spectroscopy, CD, and LSCM. The effect of different conditions on the formation and separation of COP-SC complex was determined. The extraction of the orange pectin using SC precipitation (SCOP) was performed, and the physicochemical properties of SCOP were determined and compared with the orange pectin extracted by alcohol precipitation (APOP). The results showed that the electrostatic interaction was the main interaction between these two polymers, and it was strongly dependent on pH, COP/SC ratio, and salt concentration. The mixture of COP and SC formed an electrostatic complex in the pH range of 1.5-6.8 with the absence of NaCl. The recovery rate of SCOP and precipitation rate of SC were 89.43% and 98.33% when the ratio was 1:15. The physicochemical properties of SCOP were almost the same as APOP.

Effects of orange essential oil on intestinal microflora in mice.

BACKGROUND: The intestinal microbiota has a wide variety of functions in the host. A positive effect of plant extract on intestinal microbiota in animals has been reported. However, the effect of orange essential oil and its components limonene, linalool and citral on intestinal microflora in mice has seldom been reported. The effects of intragastric administration of orange essential oil and limonene, linalool and citral on intestinal microflora and biochemical indexes in mice were studied. RESULTS: The effect of essential oil, linalool and citral on immune organ index (spleen and thymus index), IgM and IL-2 was not significant (P > 0.05). A significant increase (P < 0.05) of H+ K+ -ATPase activity, IgA, IgG, and IL-2 in the limonene group was observed when compared with the control group. Orange essential oil, limonene, linalool and citral could significantly reduce the content of short-chain fatty acids in the cecum and colon of mice. Principal coordinates analysis showed that intestinal bacterial structure of limonene group cecum and colon was apparently separated from other groups. The relative abundance of Lactobacillus in cecum and colon in essential oil, limonene, linalool and citral groups was higher than that in the control group. CONCLUSIONS: Orange essential oil and limonene, linalool and citral could affect the intestinal microflora of mice, and enhance the relative abundance of Lactobacillus. The intestinal bacterial structure of cecum and colon in the limonene group was quite different from other groups. This indicated a more obvious effect of limonene on intestinal bacteria, also resulting in significant changes in blood immune index and short-chain fatty acids in mice. © 2019 Society of Chemical Industry.

Effect of Food Emulsifiers on Aroma Release.

This study aimed to determine the influence of different emulsifiers or xanthan-emulsifier systems on the release of aroma compounds. Solid-phase microextraction (SPME) and GC-MS were used to study the effects of varying concentrations of xanthan gum, sucrose fatty acid ester, Tween 80 and soybean lecithin on the release of seven aroma compounds. The effects of the emulsifier systems supplemented with xanthan gum on aroma release were also studied in the same way. The results showed varying degrees of influence of sucrose fatty acid ester, soybean lecithin, Tween 80 and xanthan gum on the release of aroma compounds. Compared with other aroma compounds, ethyl acetate was more likely to be conserved in the solution system, while the amount of limonene released was the highest among these seven aroma compounds. In conclusion, different emulsifiers and complexes showed different surface properties that tend to interact with different aroma molecules. The present studies showed that the composition and structure of emulsifiers and specific interactions between emulsifiers and aroma molecules have significant effects on aroma release.