Dual regulation of lipid droplet-triacylglycerol metabolism and ERG9 expression for improved β-carotene production in Saccharomyces cerevisiae

Background The limitation of storage space, product cytotoxicity and the competition for precursor are the major challenges for efficiently overproducing carotenoid in engineered non-carotenogenic microorganisms. In this work, to improve β-carotene accumulation in Saccharomyces cerevisiae, a strategy that simultaneous increases cell storage capability and strengthens metabolic flux to carotenoid pathway was developed using exogenous oleic acid (OA) combined with metabolic engineering approaches. Results The direct separation of lipid droplets (LDs), quantitative analysis and genes disruption trial indicated that LDs are major storage locations of β-carotene in S. cerevisiae. However, due to the competition for precursor between β-carotene and LDs-triacylglycerol biosynthesis, enlarging storage space by engineering LDs related genes has minor promotion on β-carotene accumulation. Adding 2 mM OA significantly improved LDs-triacylglycerol metabolism and resulted in 36.4% increase in β-carotene content. The transcriptome analysis was adopted to mine OA-repressible promoters and IZH1 promoter was used to replace native ERG9 promoter to dynamically down-regulate ERG9 expression, which diverted the metabolic flux to β-carotene pathway and achieved additional 31.7% increase in β-carotene content without adversely affecting cell growth. By inducing an extra constitutive β-carotene synthesis pathway for further conversion precursor farnesol to β-carotene, the final strain produced 11.4 mg/g DCW and 142 mg/L of β-carotene, which is 107.3% and 49.5% increase respectively over the parent strain. Conclusions This strategy can be applied in the overproduction of other heterogeneous FPP-derived hydrophobic compounds with similar synthesis and storage mechanisms in S. cerevisiae. Graphical Abstract

Dual Regulation of Lipid Droplet-triacylglycerol Metabolism and ERG9 Expression for Improved β-carotene Production in Saccharomyces Cerevisiae

BackgroundThe limitation of storage space, product cytotoxicity and the competition for precursor is the major challenges for efficiently overproducing carotenoid in engineered non-carotenogenic microorganisms. In this work, to improve β-carotene accumulation in Saccharomyces cerevisiae, a strategy that simultaneous increases cell storage capability and strengthens metabolic flux to carotenoid pathway was developed using exogenous oleic acid (OA) combined with metabolic engineering approaches.ResultsThe direct separation of lipid droplets (LDs), quantitative analysis and genes disruption trial indicated that LDs are major storage locations of β-carotene in S. cerevisiae. However, due to the competition for precursor between β-carotene and LDs-triacylglycerol biosynthesis, enlarging storage space by engineering LDs related genes has minor promotion on β-carotene accumulation. Adding 2 mM OA significantly improved LDs-triacylglycerol metabolism and resulted in 36.4% increase in β-carotene content. The transcriptome analysis was adopted to mine OA-repressible promoters and IZH1 promoter was used to replace native ERG9 promoter to dynamically down-regulate ERG9 expression, which diverted the metabolic flux to β-carotene pathway and achieved additional 31.7% increase in β-carotene content without adversely affecting cell growth. By inducing an extra constitutive β-carotene synthesis pathway for further conversion precursor farnesol to β-carotene, the final strain produced 11.4 mg/DCW and 142 mg/L of β-carotene, which is 107.3% and 49.5% increase respectively over the parent strain. ConclusionsThis strategy can be applied in the overproduction of other heterogeneous FPP-derived hydrophobic compounds with similar synthesis and storage mechanisms in S. cerevisiae.

SLOW OSCILLATIONS IN DRIVES OF MACHINE ASSEMBLIES

Machine assembly with additional degrees of freedom is considered. The method of direct separation of motions was used for research. It was shown that the braking vibration moment occurring by the reason of resonance effects in the driven object can lead to excitation of rotor oscillations of the assembly with a occurrence of slow oscillations of the vibration exciter rotor is demonstrated. frequency lower than the rotation frequency. Slow rotor oscillations represent a transient process to the stationary motion mode, which is established when an additional load torque occurs. Moreover, the maximum oscillation amplitudes will be relatively large. By the example of a vibration machine with an inertial drive, the occurrence of slow oscillations of the vibration exciter rotor is demonstrated.

Th-MOF showing six-fold imide-sealed pockets for middle-size-separation of propane from nature gas

Separation of propane from nature gas is of great importance to industry. However, in light of size-based separation, there still lacks effective method to directly separate propane from nature gas, due to the comparable physical properties for these light alkanes (C1-C4) and the middle size of propane. In this work, we found that a new Th-MOF could be an ideal solution for this issue. The Th-MOF takes UiO-66-type structure, but with the pocket sealed by six-fold imide groups; this not only precisely reduces the size of pocket to exactly match propane, but also enhances the host-guest interactions through multiple supramolecular interactions. As a result, highly selective adsorption of propane over methane, ethane, and butane was observed, implying unique middle-size separation. The actual separation was confirmed by breakthrough experiments, and it is found that both relatively smaller molecules (methane and ethane) and relatively bigger molecules (butane) break through the Th-MOF column within 10 min/g, whereas propane with middle size can maintain very long retention time up to 80 min/g, strongly suggesting middle-size separation and its superior application in direct separation of propane from nature gas. The separation mechanism, as unveiled by both theoretical calculation and comparative experiments, is due to the six-fold imide-sealed pockets that could effectively distinguish propane from other light alkanes through both size effect and host-guest interactions.

Blends Containing Amphiphilic Biopolymers. Compatibility Behavior

: This mini-review deals with the miscibility behavior of two biopolymers, chitosan, and alginate. It is well known that the miscibility in multifunctional polymers blends is favored due to specific interactions, which origin a negative heat of mixing. Particular interest is focused on functionalized polymers because they are the most suitable way to obtain interacting polymers, producing a single-phase material. Due to the polyfunctionality of chitosan (CS) and other biopolymers, they can be taken into account as a basis of a strongly interacting polymer. They would allow obtaining compatible polymeric materials. For this reason, blends containing CS with different vinyl polymers have been studied. The most significant polymeric blends with these natural polymers will be analyzed in this review. Chitosan is obtained from the biopolymer chitin through sequential processes of demineralization, deproteinization, and deacetylation. The native chitin is obtained by direct separation from the marine crustaceans shell, abundant on the sea coasts. Some classic results that relate to the polymeric blends containing amphiphilic polymers will be discussed. Another biopolymer of the coast is Sodium Alginate (SA). Alginate also allows the formation of compatible polymer blends. Results in this regard will also be analyzed in this review.

The effect of high-frequency stochastic actions on the low-frequency behaviour of dynamic systems

The dynamic system described by a finite number of first-order differential equations is considered. The right side of each equation is a sum of a slow, deterministic and, in the general case, nonlinear function of dynamic variables and a stochastic excitation. The stochastic action is a superposition of a finite number of independent random processes with coefficients depending on dynamic variables and slow time. The problem statement is oriented to applications in the field of driven systems. The analysis is based on the concept of vibration mechanics proposed by I. I. Blekhman. The modified method of direct separation of slow and fast motions uses the explicit introduction of a small parameter and some ideas of the two-scale technique. The general formulas for vibrational forces (or fluxes) are obtained. These additional terms appear in the resulting system for averaged motion instead of the stochastic terms to make the averaged system equivalent to the initial stochastic system with respect to slow motions and, in particular, to low-frequency resonances. As an example, the model of a vibration machine for bulk material processing is considered. The stochastic effect is caused by random oscillations of the bulk material mass. It is transformed into a modification of the machine's frequency characteristics leading to a specific stochastic resonance. This article is part of the theme issue ‘Vibrational and stochastic resonance in driven nonlinear systems (part 1)’.

Parametric Study and Simulation of Horizontal Split Casing Pump by using Computational Fluid Dynamics

The pump requirement is required to meet the specified performance parameters means the required pressure (head) and flow, using less power. For water pump applications, periodic maintenance is required to check worn parts such as wear ring, shaft sleeves. In such a case the direct separation case pumps are reliable. If a given work point can meet the range of pumps of the available separate cases, it is necessary to develop a new pump and the upgraded pump should be more efficient with the considered power problems. With the help of computational fluid dynamics, the complex internal flow of the pump in the case of equilibrium separation can be accurately predicted, thus facilitating the construction of the pump. Simulation provides the most complete performance testing and important diagnostic details that help the engineer quickly and increase composition. As a result, the company is able to quickly create new designs such as a fully patented water pump with a patent. Pressure suction can be problematic during operation and operation of centrifugal pumps. Such pressure stimulants are traditionally investigated but quantitative analysis techniques allow these effects to be tested. Numerical analysis is done with a multi-reference framework to predict the flow field within the entire pipeline and impeller.