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1.
Specialized Plant Metabolism Characteristics and Impact on Target Molecule Biotechnological Production.
Matsuura, HN, Malik, S, de Costa, F, Yousefzadi, M, Mirjalili, MH, Arroo, R, Bhambra, AS, Strnad, M, Bonfill, M, Fett-Neto, AG
Molecular biotechnology. 2018;(2):169-183
Abstract
Plant secondary metabolism evolved in the context of highly organized and differentiated cells and tissues, featuring massive chemical complexity operating under tight environmental, developmental and genetic control. Biotechnological demand for natural products has been continuously increasing because of their significant value and new applications, mainly as pharmaceuticals. Aseptic production systems of plant secondary metabolites have improved considerably, constituting an attractive tool for increased, stable and large-scale supply of valuable molecules. Surprisingly, to date, only a few examples including taxol, shikonin, berberine and artemisinin have emerged as success cases of commercial production using this strategy. The present review focuses on the main characteristics of plant specialized metabolism and their implications for current strategies used to produce secondary compounds in axenic cultivation systems. The search for consonance between plant secondary metabolism unique features and various in vitro culture systems, including cell, tissue, organ, and engineered cultures, as well as heterologous expression in microbial platforms, is discussed. Data to date strongly suggest that attaining full potential of these biotechnology production strategies requires being able to take advantage of plant specialized metabolism singularities for improved target molecule yields and for bypassing inherent difficulties in its rational manipulation.
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2.
A review on sustainable yeast biotechnological processes and applications.
Nandy, SK, Srivastava, RK
Microbiological research. 2018;:83-90
Abstract
Yeast is very well known eukaryotic organism for its remarkable biodiversity and extensive industrial applications. Saccharomyces cerevisiae is one of the most widely used microorganisms in biotechnology with successful applications in the biochemical production. Biological conversion with the focus on the different utilization of renewable feedstocks into fuels and chemicals has been intensively investigated due to increasing concerns on sustainability issues worldwide. Compared with its counterparts, Saccharomyces cerevisiae, the baker's yeast, is more industrially relevant due to known genetic and physiological background, the availability of a large collection of genetic tools, the compatibility of high-density and large-scale fermentation, and optimize the pathway for variety of products. Therefore, S. cerevisiae is one of the most popular cell factories and has been successfully used in the modern biotech industry to produce a wide variety of products such as ethanol, organic acids, amino acids, enzymes, and therapeutic proteins. This study explores how different sustainable solutions used to overcome various environmental effects on yeast. This work targets a broad matrix of current advances and future prospect in yeast biotechnology and discusses their application and potential in general.
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3.
Arabidopsis: the original plant chassis organism.
Holland, CK, Jez, JM
Plant cell reports. 2018;(10):1359-1366
Abstract
Arabidopsis thaliana (thale cress) has a past, current, and future role in the era of synthetic biology. Arabidopsis is one of the most well-studied plants with a wealth of genomics, genetics, and biochemical resources available for the metabolic engineer and synthetic biologist. Here we discuss the tools and resources that enable the identification of target genes and pathways in Arabidopsis and heterologous expression in this model plant. While there are numerous examples of engineering Arabidopsis for decreased lignin, increased seed oil, increased vitamins, and environmental remediation, this plant has provided biochemical tools for introducing Arabidopsis genes, pathways, and/or regulatory elements into other plants and microorganisms. Arabidopsis is not a vegetative or oilseed crop, but it is as an excellent model chassis for proof-of-concept metabolic engineering and synthetic biology experiments in plants.
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4.
Direct Photosynthetic Production of Plastic Building Block Chemicals from CO2.
Song, X, Wang, Y, Diao, J, Li, S, Chen, L, Zhang, W
Advances in experimental medicine and biology. 2018;:215-238
Abstract
Hydroxy acids have attracted attention as building block chemicals due to their roles as precursors for the production of various pharmaceuticals, vitamins, antibiotics, and flavor compounds as well as monomers for biodegradable plastic polyesters. The current approach to hydroxy acid production relies on nonrenewable fossil resources such as petroleum for raw materials, raising issues such as the rising costs of starting materials and environmental incompatibility. Recently, synthetic biology approaches based on the rational design and reconstruction of new biological systems were implemented to produce chemicals from a variety of renewable substrates. In addition to research using heterotrophic organic carbon-dependent Escherichia coli or yeasts, photosynthetic microorganisms such as cyanobacteria possessing the ability to absorb solar radiation and fix carbon dioxide (CO2) as a sole carbon source have been engineered into a new type of microbial cell factory to directly produce hydroxy acids from CO2. In this chapter, recent progress regarding the direct photosynthetic production of three important hydroxy acids-3-hydroxypropionate (3-HP), 3-hydroxybutyrate (3-HB), and 3-hydroxyvalerate (3-HV)-from CO2 in cyanobacteria is summarized and discussed.
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5.
Production of plant-derived polyphenols in microorganisms: current state and perspectives.
Milke, L, Aschenbrenner, J, Marienhagen, J, Kallscheuer, N
Applied microbiology and biotechnology. 2018;(4):1575-1585
Abstract
Plants synthesize several thousand different polyphenols of which many have the potential to aid in preventing or treating cancer, cardiovascular, and neurodegenerative diseases. However, plants usually contain complex polyphenol mixtures impeding access to individual compounds in larger quantities. In contrast, functional integration of biosynthetic plant polyphenol pathways into microorganisms allows for the production of individual polyphenols as chemically distinct compounds, which can be synthesized in large amounts and can be more easily isolated. Over the last decade, microbial synthesis of many plant polyphenols could be achieved, and along the way, many decisive bottlenecks in the endogenous microbial host metabolism as well as in the heterologous plant pathways could be identified. In this review, we present recent advancements in metabolic engineering of microorganisms for the production of plant polyphenols and discuss how current challenges could be addressed in the future.
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6.
Metabolic engineering of Pichia pastoris.
Peña, DA, Gasser, B, Zanghellini, J, Steiger, MG, Mattanovich, D
Metabolic engineering. 2018;:2-15
Abstract
Besides its use for efficient production of recombinant proteins the methylotrophic yeast Pichia pastoris (syn. Komagataella spp.) has been increasingly employed as a platform to produce metabolites of varying origin. We summarize here the impressive methodological developments of the last years to model and analyze the metabolism of P. pastoris, and to engineer its genome and metabolic pathways. Efficient methods to insert, modify or delete genes via homologous recombination and CRISPR/Cas9, supported by modular cloning techniques, have been reported. An outstanding early example of metabolic engineering in P. pastoris was the humanization of protein glycosylation. More recently the cell metabolism was engineered also to enhance the productivity of heterologous proteins. The last few years have seen an increased number of metabolic pathway design and engineering in P. pastoris, mainly towards the production of complex (secondary) metabolites. In this review, we discuss the potential role of P. pastoris as a platform for metabolic engineering, its strengths, and major requirements for future developments of chassis strains based on synthetic biology principles.
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7.
MiYA, an efficient machine-learning workflow in conjunction with the YeastFab assembly strategy for combinatorial optimization of heterologous metabolic pathways in Saccharomyces cerevisiae.
Zhou, Y, Li, G, Dong, J, Xing, XH, Dai, J, Zhang, C
Metabolic engineering. 2018;:294-302
Abstract
Facing boosting ability to construct combinatorial metabolic pathways, how to search the metabolic sweet spot has become the rate-limiting step. We here reported an efficient Machine-learning workflow in conjunction with YeastFab Assembly strategy (MiYA) for combinatorial optimizing the large biosynthetic genotypic space of heterologous metabolic pathways in Saccharomyces cerevisiae. Using β-carotene biosynthetic pathway as example, we first demonstrated that MiYA has the power to search only a small fraction (2-5%) of combinatorial space to precisely tune the expression level of each gene with a machine-learning algorithm of an artificial neural network (ANN) ensemble to avoid over-fitting problem when dealing with a small number of training samples. We then applied MiYA to improve the biosynthesis of violacein. Feed with initial data from a colorimetric plate-based, pre-screened pool of 24 strains producing violacein, MiYA successfully predicted, and verified experimentally, the existence of a strain that showed a 2.42-fold titer improvement in violacein production among 3125 possible designs. Furthermore, MiYA was able to largely avoid the branch pathway of violacein biosynthesis that makes deoxyviolacein, and produces very pure violacein. Together, MiYA combines the advantages of standardized building blocks and machine learning to accelerate the Design-Build-Test-Learn (DBTL) cycle for combinatorial optimization of metabolic pathways, which could significantly accelerate the development of microbial cell factories.
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8.
Corynebacterium glutamicum for Sustainable Bioproduction: From Metabolic Physiology to Systems Metabolic Engineering.
Becker, J, Gießelmann, G, Hoffmann, SL, Wittmann, C
Advances in biochemical engineering/biotechnology. 2018;:217-263
Abstract
Since its discovery 60 years ago, Corynebacterium glutamicum has evolved into a workhorse for industrial biotechnology. Traditionally well known for its remarkable capacity to produce amino acids, this Gram-positive soil bacterium, has become a flexible, efficient production platform for various bulk and fine chemicals, materials, and biofuels. The central turnstile of all these achievements is our excellent understanding of its metabolism and physiology. This knowledge base, together with innovative systems metabolic engineering concepts, which integrate systems and synthetic biology into strain engineering, has upgraded C. glutamicum into one of the most successful industrial microorganisms in the world.
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9.
Transformation techniques for metabolic engineering of diatoms and haptophytes: current state and prospects.
Velmurugan, N, Deka, D
Applied microbiology and biotechnology. 2018;(10):4255-4267
Abstract
Diatoms and haptophytes represent a key segment of the dominant phytoplankton communities that frequently form massive blooms in the photic zone of the ocean and are considered indicators of global climate changes. Diatoms and haptophytes also play a vital role in the biological carbon fixation in the carbon cycles. Carbon partitioning within diatoms and haptophytes possesses a wide range of chemical compounds and storage materials, such as lipids, carbohydrates, and chlorophyll. Among the marine microorganisms, diatoms and haptophytes have been recognized as promising sources of long- and very long-chain polyunsaturated fatty acids (PUFA). So far, a variety of approaches have been employed for genetic modification in the nuclei of diatoms and haptophytes. Studies on transformation and metabolic engineering in various intracellular genomes, such as chloroplast and mitochondria, are scarce. Particle bombardment, Agrobacterium and PEG-mediated gene transfer, and electroporation have been reported for foreign gene transformation into the diatoms and haptophytes. Antibiotics (G418 and chloramphenicol) and herbicides (zeocin, hygromycin, and norflurazon) have been successfully demonstrated as the best selection markers. Despite the availability of a wide range of molecular tools for foreign gene expression in microalgae, very few promoters (lhcf1, nr, h4, ef2, fcp, and pds) have been reported for diatoms and haptophytes. Therefore, in this review, we first summarize the significant progress that has been achieved in transgene expression in diatoms and haptophytes and highlight the importance and availability of recently developed novel tools that are suitable for transgenic expression in diatoms and haptophytes.
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10.
Can the world's favorite fruit, tomato, provide an effective biosynthetic chassis for high-value metabolites?
Li, Y, Wang, H, Zhang, Y, Martin, C
Plant cell reports. 2018;(10):1443-1450
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Abstract
Tomato has a relatively short growth cycle (fruit ready to pick within 65-85 days from planting) and a relatively high yield (the average for globe tomatoes is 3-9 kg fruit per plant rising to as much as 40 kg fruit per plant). Tomatoes also produce large amounts of important primary and secondary metabolites which can serve as intermediates or substrates for producing valuable new compounds. As a model crop, tomato already has a broad range of tools and resources available for biotechnological applications, either increased nutrients for health-promoting biofortified foods or as a production system for high-value compounds. These advantages make tomato an excellent chassis for the production of important metabolites. We summarize recent achievements in metabolic engineering of tomato and suggest new candidate metabolites which could be targets for metabolic engineering. We offer a scheme for how to establish tomato as a chassis for industrial-scale production of high-value metabolites.