Untargeted Global Metabolomic Analysis Reveals the Mechanism of Tripropylamine-Enhanced Lycopene Accumulation in Blakeslea trispora

We previously determined that the cyclase inhibitor tripropylamine (TPA) significantly enhances lycopene accumulation in Blakeslea trispora. To elucidate the mechanism of TPA-enhanced lycopene accumulation, the untargeted metabolome of B. trispora treated with TPA was analyzed by UHPLC-Q-TOF/MS. Forty-two differential metabolites were identified, of which 15 significantly differential metabolites meeting the following parameters were screened: variable importance for the projection > 1, P < 0.05, and fold change > 1.5. The down-regulated metabolites were mainly cyclic dipeptides, bacteriostatic compounds, and lipids, while the up-regulated metabolites were mainly unsaturated fatty acid. Furthermore, the bacteriostatic ability was poor, the extracellular and intracellular pH levels were high, and hyphae with vesicles were swollen locally in B. trispora after treatment with TPA. Our data suggest that the TPA enhances lycopene accumulation not only by inhibiting the cyclization of β-carotene but also by down-regulating cyclic dipeptides for quorum sensing; up-regulating unsaturated fatty acids, 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine, and 4-hydroxybenzoate and down-regulating choline, resulting in locally swelling mycelium with vacuoles; and down-regulating bacteriostatic metabolites for metabolic flux redistribution.

The Inhibitory Effect of Cyclodextrin on Oxygen Bioavailability Is a Key Factor for the Metabolic Flux Redistribution Towards Steroid Alcohols in Phytosterol Resting Cells Bioconversion

In the present work, we tried to identify the mechanism why by which the steroid alcohols accumulated when hydroxypropyl-β-cyclodextrin (HP-β-CD) was present to enhance the sterol conversion rate. Compared with the bioconversion system without HP-β-CD, the reaction rate was greatly improved in presence of HP-β-CD, but the steroid alcohols largely accumulated concurrently. The concentration of steroid alcohols increased with the enhanced reaction rate and the higher intracellular NADH/NAD+ level was detected. Mycobacterium neoaurum mutants with higher KshA activity (3-ketosteroid 9α-hydrolase, a monooxygenase hydroxylating the nucleus at C-9 at the expense of NAD(P)H consumption) reduced the steroid alcohols production and in the meantime, the NADH/NAD+ level was decreased consequently. Further research found that oxygen availability was seriously inhibited by the cyclodextrin in a reaction system. These results indicated that NADH formed in the bioconversion was not properly regenerated via the respiratory chain because of the poor oxygen bioavailability. The inhibitory effect of cyclodextrin on oxygen bioavailability is a key factor for the metabolic flux redistribution towards steroid alcohols in phytosterol resting cells bioconversion.

Optimization of In-Stage Diversion To Promote Uniform Planar Multifracture Propagation: A Numerical Study

Summary Creating uniform multiple fractures is a challenging task due to reservoir heterogeneity and stress shadow. Limited-entry perforation and in-stage diversion are commonly used to improve multifracture treatments. Many studies have investigated the mechanism of limited-entry perforation for multifracture treatments, but relatively few have focused on the in-stage diversion process. The design of in-stage diversion is usually through trial and error because of the lack of a simulator. In this study, we present a fully coupled planar 2D multifracture model for simulating the in-stage diversion process. The objective is to evaluate flux redistribution after diversion and optimize the dosage of diverters and diversion timing under different in-stage in-situ stress difference. Our model considers ball sealer allocation and solves flux redistribution after diversion through a fully coupled multifracture model. A supertimestepping explicit algorithm is adopted to solve the solid/fluid coupling equations efficiently. Multifracture fronts are captured by using tip asymptotes and an adaptive time-marching approach. The modeling results are validated against analytical solutions for a plane-strain Khristianovic-Geertsma de Klerk (KGD) model. A series of numerical simulations are conducted to investigate the multifracture growth under different in-stage diversion operations. Parametric studies reveal that the in-stage in-situ stress difference is a critical parameter for diversion designs. When the in-situ stress difference is larger than 2 MPa, the fracture in the high-stress zone can hardly be initiated before diversion for a general fracturing design. More ball sealers are required for the formations with higher in-stage in-situ stress difference. The diverting time should be earlier for formations with high in-stage stress differences as well. Adding more perforation holes in the zone with higher in-situ stress is recommended to achieve even flux distribution. The results of this study can help understand the multifracture growth mechanism during in-stage diversion and optimize the diversion design timely.

Distorted surfaces of magnetic helium-peculiar stars: an application to a Cen

ABSTRACT Helium-peculiar magnetic chemically peculiar stars show variations of helium abundance across their surfaces. As a result of associated atmospheric scale height variations, the stellar surface becomes distorted, with helium-rich regions dented inwards. Effectively, on top of flux variations due to opacity effects, the depressed helium-rich surface regions become less bright in the optical regions and brighter in the ultraviolet. We study the observational effects of the aspherical surface on the light curves of a Cen. We simulate the light curves of this star adopting surface distributions of He, N, O, Si, and Fe derived from Doppler mapping and introducing the effect of distortion proportional to helium abundance. We show that while most of the optical and UV variations of this star result from flux redistribution due to the non-uniform surface distributions of helium and iron, the reduction of light variations due to the helium-related surface distortion leads to a better agreement between simulated optical light curves and the light curves observed with the BRITE satellites.