Transcriptomics analysis and fed-batch regulation of high astaxanthin-producing Phaffia rhodozyma/Xanthophyllomyces dendrorhous obtained through adaptive laboratory evolution.

Astaxanthin has high utilization value in functional food because of its strong antioxidant capacity. However, the astaxanthin content of Phaffia rhodozyma is relatively low. Adaptive laboratory evolution is an excellent method to obtain high-yield strains. TiO2 is a good inducer of oxidative stress. In this study, different concentrations of TiO2 were used to domesticate P. rhodozyma, and at a concentration of 1000 mg/L of TiO2 for 105 days, the optimal strain JMU-ALE105 for astaxanthin production was obtained. After fermentation, the astaxanthin content reached 6.50 mg/g, which was 41.61% higher than that of the original strain. The ALE105 strain was fermented by batch and fed-batch, and the astaxanthin content reached 6.81 mg/g. Transcriptomics analysis showed that the astaxanthin synthesis pathway, and fatty acid, pyruvate, and nitrogen metabolism pathway of the ALE105 strain were significantly upregulated. Based on the nitrogen metabolism pathway, the nitrogen source was adjusted by ammonium sulphate fed-batch fermentation, which increased the astaxanthin content, reaching 8.36 mg/g. This study provides a technical basis and theoretical research for promoting industrialization of astaxanthin production of P. rhodozyma. ONE-SENTENCE SUMMARY: A high-yield astaxanthin strain (ALE105) was obtained through TiO2 domestication, and its metabolic mechanism was analysed by transcriptomics, which combined with nitrogen source regulation to further improve astaxanthin yield.

The identification of biotransformation pathways for removing fishy malodor from Bangia fusco-purpurea using fermentation with Saccharomyces cerevisiae.

The yeast Saccharomyces cerevisiae is effective in reducing the fishy malodor of sea products. However, the biotransformation pathways are still unclear. The seaweed B. fusco-purpurea was taken as an example to investigate the chemical transformation pathways for the deodorization process with S. cerevisiae fermentation. Sensory evaluation, GC-MS, GC-MS-O and odor activity value (OAV) analyses showed the fishy odorants were 1-octen-3-ol, (E)-2-nonenal, 2,4-decadienal, 2-pentylfuran, 2-octen-1-ol and nonanal. The removal of fishy malodor was related to the reactions of reduction, dehydrogenation, deformylation-oxygenation and ester syntheses via catalysis of aldehyde dehydrogenase, alcohol dehydrogenases, epoxide hydrolase, aldehyde deformylating-oxygenase, enone reductase, oxidases, dehydrogenases, aldo-keto reductases, ester synthase and acyltransferase. Interestingly, for the first time, it was found that 3,5-octadien-2-one transformed to 6-octen-2-one; and 2-pentylfuran transformed to o-cymene and hexyl acetate. Our findings enrich the knowledge for the removal of fishy malodor from sea products such as seaweeds.

Preparation of isoquercitrin by biotransformation of rutin using α-L-rhamnosidase from Aspergillus niger JMU-TS528 and HSCCC purification.

Isoquercitrin is a flavonoid with important applications in the pharmaceutical and nutraceutical industries. However, a low yield and high production cost hinders the industrial preparation of isoquercitrin. In the present study, isoquercitrin was prepared by biotransformation of rutin using α-L-rhamnosidase from Aspergillus niger JMU-TS528 combined with high-speed countercurrent chromatography (HSCCC) purification. As a result, the optimum transformation pH, temperature, and time were pH 4.0, 60 °C, and 60 min, respectively. The Km and Vmax were 0.36 mM and 0.460 mmol/min, respectively. For isoquercitrin production, the optimal rutin concentration and transformation time were approximately 1000 mg/L and 60 min. The rutin transformation rate reached 96.44%. The isoquercitrin was purified to a purity of 97.83% using one-step purification with HSCCC. The isoquercitrin was identified using UPLC-Q-TOF-MS. The comprehensive results indicated that the biotransformation procedure using the α-L-rhamnosidase from A. niger JMU-TS528 combined with HSCCC was a simple and effective process to prepare isoquercitrin, which might facilitate the production of isoquercitrin for industrial use.

Antioxidant capacity and prebiotic effects of Gracilaria neoagaro oligosaccharides prepared by agarase hydrolysis.

Gracilaria is a genus of red algae widely cultivated in Asia and is notable for its economic importance as food ingredients. Neoagaro oligosaccharides (NAOSs) are products of Gracilaria that have excellent water solubility, antioxidant activity and prebiotic effect. In this study, Gracilaria crude polysaccharide was treated with agarase and hydrolyzed into NAOSs with different degrees of polymerization (DP). The compositions of the hydrolyzed NAOSs were analyzed by electrospray ionization-time of flight-mass spectrometry and thin layer chromatography. The antioxidant capacity and prebiotic effects of NAOSs with different DPs were investigated and the results showed that DP could affect the antioxidant capacity of NAOSs. The prebiotic effects of NAOSs with different DP were evaluated based on the influence on the growth of four intestinal bacteria. NAOSs promoted the growth of Lactobacillus delbrueckii subsp. bulgaricus and Sterptococcus thermophilus. The protective effect of Gracilaria NAOSs in simulated gastrointestinal juice was also studied. Finally, NAOSs with best prebiotic effects were used in Procambarus feeding experiment and exhibited promotion effect on Procambarus growth. The present study showed that Gracilaria NAOSs can be utilized as antioxidant and prebiotic additive, which had a considerable potential in food and feed industry.

Water accelerated transformation of d-limonene induced by ultraviolet irradiation and air exposure.

d-Limonene is a fragrant chemical that widely exists in aromatic products. Isotopic labelling of water molecules plus GC-MS and GC-PCI-Q-TOF analyses were used to investigate the influence of water molecules on chemical transformation of d-limonene induced by UV irradiation and air exposure. The results showed that the synergistic effect of UV irradiation, air exposure and water presence could facilitate d-limonene transformation into the limonene oxides: p-mentha-2,8-dienols, hydroperoxides, carveols, l-carvone and carvone oxide. UV irradiation, air exposure, or water alone, however, caused negligible d-limonene transformation. With the aid of isotopic labelling of water and oxygen molecules, it was found that water molecules were split into hydrogen radicals and hydroxyl radicals, and the hydrogen radicals, in particular, promoted the transformation reactions. This study has elucidated the mechanism and factors that influence the transformation of d-limonene, which will benefit industries involved in production and storage of d-limonene-containing products.

Recovery and purification of limonin from pummelo [Citrus grandis] peel using water extraction, ammonium sulfate precipitation and resin adsorption.

Limonin is a bioactive compound that is traditionally extracted from citrus seeds using organic solvents or alkaline/metal ion solutions. In the present study, pummelo [Citrus grandis] peel was investigated for limonin preparation using a novel process consisting of water extraction, ammonium sulfate precipitation and resin adsorption. The pummelo peel was determined to have 4.7mg/g limonin, which could be extracted by water and further recovered by ammonium sulfate precipitation with a yield of 2.4mg/g, which was similar to that of traditional process using ethanol extraction and vacuumed evaporation. The precipitated limonin was purified by resin adsorption and crystallization with a purity of 96.4%. In addition, the limonin was identified via the analyses of retention time, infrared spectrum and nuclear magnetic resonance. This study indicates a novel and eco-friendly process for recovering limonin, providing a new candidate for limonin preparation.