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Phaeodactylum tricornutum: A Diatom Cell Factory
A switch from a petroleum-based to a biobased economy requires the capacity to produce both high-value low-volume and low-value high-volume products. Recent evidence supports the development of microalgae-based microbial cell factories with the objective of establishing environmentally sustainable manufacturing solutions. Diatoms display rich diversity and potential in this regard. We focus on Phaeodactylum tricornutum, a pennate diatom that is commonly found in marine ecosystems, and discuss recent trends in developing the diatom chassis for the production of a suite of natural and genetically engineered products. Both upstream and downstream developments are reviewed for the commercial development of P. tricornutum as a cell factory for a spectrum of marketable products.
P. tricornutum CCAP 1055/1 is a diatom with a sequenced annotated genome with >100 000 expression sequence tags. Transformation is becoming routine using biolistics, electroporation, and more recently by conjugation with E. coli. There is improved understanding of its biochemical pathways.P. tricornutum is showcased as a microalgal cell factory with a product spectrum including eicosapentaenoic acid, fucoxanthin, neutral lipids, and crysolaminarin, all producible with the chassis, in addition to heterologous recombinant proteins, triterpenoids (lupeol and betulin), and bioplastics precursors.P. tricornutum can be readily harvested with a host of approaches, and relevant products can be recovered using green extraction processes such as microwave-assisted and pressurized liquid extraction, with the potential to develop sustainable cost-effective processes in a biorefinery approach.
Journal Article
Microbial Chassis Development for Natural Product Biosynthesis
Engineering microbial cells to efficiently synthesize high-value-added natural products has received increasing attention in recent years. In this review, we describe the pipeline to build chassis cells for natural product production. First, we discuss recently developed genome mining strategies for identifying and designing biosynthetic modules and compare the characteristics of different host microbes. Then, we summarize state-of-the-art systems metabolic engineering tools for reconstructing and fine-tuning biosynthetic pathways and transport mechanisms. Finally, we discuss the future prospects of building next-generation chassis cells for the production of natural products. This review provides theoretical guidance for the rational design and construction of microbial strains to produce natural products.
Recent advances in omics, in silico modeling analysis and design, and DNA assembly provide big data and various tools to identify, design, and assemble the synthesis modules of natural products.Besides classical strains, various other microorganisms can be used as chassis cells for natural products due to developments in systems biology and synthetic biology.Metabolic engineering based on genetic circuits and novel genome editing tools can optimize the complex pathway of natural products.Biosensor-based high-throughput screening helps to identify transporters for natural products and facilitate their secretion.
Journal Article
NVH Investigation of Vehicle Chassis
This study presents a method for enhancing vehicle design through the integration of CAD-based topology optimization and finite element analysis (FEA) simulation. Specifically, the vehicle chassis is refined by incorporating design elements inspired by biomimicry, which are integrated to improve the vehicle’s NVH (Noise, Vibration, Harshness) performance. Noise and vibration have consistently played a critical role in vehicle design, as such, automakers devote substantial attention to minimizing the transfer of road and engine-induced disturbances into the cabin. This reduction is crucial, as excessive noise and vibration can lead to an unpleasant driving experience and may accelerate driver fatigue. Moreover, if the chassis does not effectively absorb these disturbances, the resulting stress on vehicle components can be significantly increased.In this study, FEA is employed to establish a baseline chassis model, with simulations conducted to analyse its natural vibrational modal response. Once the initial performance thresholds are established, identical constraints are applied to assess the vibrational response of the modified chassis featuring the optimized structure. The results underscore the critical value of integrating CAD and FEA methodologies in vehicle design, demonstrating a substantial positive impact by reducing development time and enhancing the overall quality of the resulting prototypes.
Journal Article
Synthetic Biology Toolbox and Chassis Development in Bacillus subtilis
Based on technical advances in the sequencing and synthesis of genetic components as well as the genome, significant progress has recently been made in developing synthetic biology toolboxes and chassis for the model Gram-positive bacterium Bacillus subtilis. In this review, we discuss recently developed synthetic biology toolboxes, including gene expression toolsets and genome editing tools. Next, advances in the B. subtilis chassis and its applications are discussed in comparison to those of other model microorganisms. Finally, future directions for the integrative use of B. subtilis synthetic biology tools and the development of an advanced chassis for efficient biomanufacturing are discussed. These factors are expected to become a major driving force for facilitating biotechnological applications of B. subtilis.
Recent development of synthetic biology toolboxes for B. subtilis, including gene expression regulatory toolboxes and genome-wide editing tools, provides powerful tools for precise gene expression control and efficient genome editing.
Advances of B. subtilis chassis and their applications help to understand fundamental cellular processes and techniques for improving production of biomolecules or heterologous enzymes.
Comparing B. subtilis chassis development with E. coli and S. cerevisiae chassis may provide potential directions for B. subtilis chassis construction.
Journal Article
Impact on the Operation of a Forwarder with the Wheeled, Tracked-Wheel or Tracked Chassis on the Soil Surface
The impact of a small forwarder with the wheeled chassis, tracked-wheel chassis, and tracked chassis traveling on the soil profile was studied. The three chassis types were assessed for the influence of the loading of forwarder cargo space and the degree of tire inflation on induced specific and actual pressures of tires on the soil surface. Penetrometric resistances of soil profile and rut depths in the forwarder driving track were measured. The effect of a layer of logging residues in the forwarder driving track on the size of induced actual pressures was determined. The practice of determining the impact of forest machines on the soil surface by means of a specific tire pressure does not have a full informative value. In the forwarder wheeled chassis, maximum values of actual pressures exceeded specific pressures established numerically by up to 203%. Average values of actual pressures could be reduced by 45% by reducing the pressure of tire inflation, by 70% with the use of tracks, or by 49% by traveling on the layer of logging residues. As compared with the wheeled chassis type, the tracked type of the forwarder chassis induced actual pressures to lower by 81% and the rut depth after ten forwarder passes was smaller by 50%.
Journal Article
Design and Analysis of Composite Structures for Automotive Applications - Chassis and Drivetrain
This book provides a theoretical background for the development of elements of car suspensions. It begins with a description of the elastic-kinematics of the vehicle and closed form solutions for the vertical and lateral dynamics. It evaluates the vertical, lateral, and roll stiffness of the vehicle, and explains the necessity of the modelling of the vehicle stiffness. The composite materials for the suspension and powertrain design are discussed and their mechanical properties are provided. The book also looks at the basic principles for the design optimization using composite materials and mass reduction principles. Additionally, references and conclusions are presented in each chapter. This book offers complete coverage of chassis components made of composite materials and covers elastokinematics and component compliances of vehicles. It looks at parts made of composite materials such as stabilizer bars, wheels, half-axes, springs, and semi-trail axles. The book also provides information on leaf spring assembly for motor vehicles and motor vehicle springs comprising composite materials.
eBook
Rational construction of genome-reduced and high-efficient industrial Streptomyces chassis based on multiple comparative genomic approaches
Background
Streptomyces chattanoogensis
L10 is the industrial producer of natamycin and has been proved a highly efficient host for diverse natural products. It has an enormous potential to be developed as a versatile cell factory for production of heterologous secondary metabolites. Here we developed a genome-reduced industrial
Streptomyces chassis
by rational ‘design-build-test’ pipeline.
Results
To identify candidate large non-essential genomic regions accurately and design large deletion rationally, we performed genome analyses of
S. chattanoogensis
L10 by multiple computational approaches, optimized Cre/
loxP
recombination system for high-efficient large deletion and constructed a series of universal suicide plasmids for rapid
loxP
or
loxP
mutant sites inserting into genome. Subsequently, two genome-streamlined mutants, designated
S. chattanoogensis
L320 and L321, were rationally constructed by depletion of 1.3 Mb and 0.7 Mb non-essential genomic regions, respectively. Furthermore, several biological performances like growth cycle, secondary metabolite profile, hyphae morphological engineering, intracellular energy (ATP) and reducing power (NADPH/NADP
+
) levels, transformation efficiency, genetic stability, productivity of heterologous proteins and secondary metabolite were systematically evaluated. Finally, our results revealed that L321 could serve as an efficient
chassis
for the production of polyketides.
Conclusions
Here we developed the combined strategy of multiple computational approaches and site-specific recombination system to rationally construct genome-reduced
Streptomyces
hosts with high efficiency. Moreover, a genome-reduced industrial
Streptomyces chassis
S. chattanoogensis
L321 was rationally constructed by the strategy, and the
chassis
exhibited several emergent and excellent performances for heterologous expression of secondary metabolite. The strategy could be widely applied in other
Streptomyces
to generate miscellaneous and versatile
chassis
with minimized genome. These
chassis
can not only serve as cell factories for high-efficient production of valuable polyketides, but also will provide great support for the upgrade of microbial pharmaceutical industry and drug discovery.
Journal Article
Topology optimization of electric vehicle chassis structure with distributed load-bearing batteries
This paper presents a systematic design approach of conceptually forming a lightweight electric vehicle (EV) chassis topology integrated with distributed load-bearing batteries of different shapes and dimensions using a density-based topology optimization approach. A deformable feature description function tailored to commercial Li-ion batteries is proposed to describe cell features with desirable layouts, dimensions, and continuous shapes only from cylinder to cube by applying a handful of design variables. A Kreisselmeier–Steinhauser function Boolean operation and a gradient-norm method are sequentially leveraged to integrate multiple cells enclosed by reinforced shells into a unified battery set. Besides, a new non-overlapping constraint is developed to avoid the geometric overlaps between all cells and further restrict the minimum battery spacing through introducing an auxiliary density filter. By solving the optimization problem, an EV chassis with distributed various specification batteries can be obtained, which exhibits better comprehensive mechanical properties than that with centralized uniform specification batteries under the same battery capacity and structural weight. Numerical examples of different battery capacity requirements, battery shell thicknesses, and minimum battery spacing are given to demonstrate the applicability of the proposed approach.
Journal Article
Industrial biotechnology of Pseudomonas putida: advances and prospects
Pseudomonas putida is a Gram-negative, rod-shaped bacterium that can be encountered in diverse ecological habitats. This ubiquity is traced to its remarkably versatile metabolism, adapted to withstand physicochemical stress, and the capacity to thrive in harsh environments. Owing to these characteristics, there is a growing interest in this microbe for industrial use, and the corresponding research has made rapid progress in recent years. Hereby, strong drivers are the exploitation of cheap renewable feedstocks and waste streams to produce value-added chemicals and the steady progress in genetic strain engineering and systems biology understanding of this bacterium. Here, we summarize the recent advances and prospects in genetic engineering, systems and synthetic biology, and applications of P. putida as a cell factory.Key points• Pseudomonas putida advances to a global industrial cell factory.• Novel tools enable system-wide understanding and streamlined genomic engineering.• Applications of P. putida range from bioeconomy chemicals to biosynthetic drugs.
Journal Article