Recent Advances on Feasible Strategies for Monoterpenoid Production in Saccharomyces cerevisiae

Terpenoids are a large diverse group of natural products which play important roles in plant metabolic activities. Monoterpenoids are the main components of plant essential oils and the active components of some traditional Chinese medicinal herbs. Some monoterpenoids are widely used in medicine, cosmetics and other industries, and they are mainly obtained by plant biomass extraction methods. These plant extraction methods have some problems, such as low efficiency, unstable quality, and high cost. Moreover, the monoterpenoid production from plant cannot satisfy the growing monoterpenoids demand. The development of metabolic engineering, protein engineering and synthetic biology provides an opportunity to produce large amounts of monoterpenoids eco-friendly using microbial cell factories. This mini-review covers current monoterpenoids production using Saccharomyces cerevisiae. The monoterpenoids biosynthetic pathways, engineering of key monoterpenoids biosynthetic enzymes, and current monoterpenoids production using S. cerevisiae were summarized. In the future, metabolically engineered S. cerevisiae may provide one possible green and sustainable strategy for monoterpenoids supply.

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Efficient Conversion of Glycerol to Ethanol by an Engineered Saccharomyces cerevisiae Strain

Applied and Environmental Microbiology ◽

10.1128/aem.00268-21 ◽

2021 ◽

Author(s):

Sadat M. R. Khattab ◽

Takashi Watanabe

Keyword(s):

Saccharomyces Cerevisiae ◽

Metabolic Flux ◽

Triosephosphate Isomerase ◽

Plant Biomass ◽

Agricultural Waste ◽

Glycerol Dehydrogenase ◽

Cell Wall Polysaccharides ◽

Plant Oil ◽

Cell Factories ◽

Engineered Strain

Glycerol is an eco-friendly solvent that enhances plant biomass decomposition via glycerolysis in many pretreatment methods. Nonetheless, inefficient conversion of glycerol to ethanol by natural Saccharomyces cerevisiae limits its use in these processes. Here, we have developed an efficient glycerol-converting yeast strain by genetically modifying the oxidation of cytosolic nicotinamide adenine dinucleotide (NADH) by an O 2 -dependent dynamic shuttle and abolishing both glycerol phosphorylation and biosynthesis in S. cerevisiae D452-2 strain, as well as vigorous expression of whole genes in the DHA-pathway ( Candid utilis glycerol facilitator, Ogataea polymorpha glycerol dehydrogenase, endogenous dihydroxyacetone kinase, and triosephosphate isomerase). The engineered strain showed conversion efficiencies (CE) up to 0.49 g ethanol/g glycerol (98% of theoretical CE), with production rate >1 g/L −1 h −1 when glycerol was supplemented in a single fed-batch fermentation in a rich medium. Furthermore, the engineered strain converted a mixture of glycerol and glucose into bioethanol (>86 g/L) with 92.8% CE. To the best of our knowledge, this is the highest reported titer of bioethanol produced from glycerol and glucose. Notably, we developed a glycerol-utilizing transformant from parent strain, which cannot utilize glycerol as a sole carbon source. The developed strain converted glycerol to ethanol with a productivity of 0.44 g/L −1 h −1 on minimal medium under semi-aerobic conditions. Our findings will promote the utilization of glycerol in eco-friendly biorefineries and integrate bioethanol and plant-oil industries. IMPORTANCE With the development of efficient lignocellulosic biorefineries, glycerol has attracted attention as an eco-friendly biomass-derived solvent that can enhance the dissociation of lignin and cell wall polysaccharides during the pretreatment process. Co-conversion of glycerol with the sugars released from biomass after glycerolysis increases the resources for ethanol production and lowers the burden of component separation. However, low conversion efficiency from glycerol and sugars limits the industrial application of this process. Therefore, the generation of an efficient glycerol-fermenting yeast will promote the applicability of integrated biorefineries. Hence, metabolic flux control in yeast grown on glycerol will lead to the generation of cell factories that produce chemicals, which will boost biodiesel and bioethanol industries. Additionally, the use of glycerol-fermenting yeast will reduce global warming and generation of agricultural waste, leading to the establishment of a sustainable society.

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Microbial Cell Factories for Green Production of Vitamins

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.661562 ◽

2021 ◽

Vol 9 ◽

Author(s):

Yanyan Wang ◽

Linxia Liu ◽

Zhaoxia Jin ◽

Dawei Zhang

Keyword(s):

Water Soluble ◽

Microbial Cell ◽

Synthesis Methods ◽

Renewable Chemicals ◽

Microbial Cell Factories ◽

Cell Factories ◽

Green Production ◽

Optimal Health ◽

Metabolic Activities ◽

Fat Soluble Vitamins

Vitamins are a group of essential nutrients that are necessary to maintain normal metabolic activities and optimal health. There are wide applications of different vitamins in food, cosmetics, feed, medicine, and other areas. The increase in the global demand for vitamins has inspired great interest in novel production strategies. Chemical synthesis methods often require high temperatures or pressurized reactors and use non-renewable chemicals or toxic solvents that cause product safety concerns, pollution, and hazardous waste. Microbial cell factories for the production of vitamins are green and sustainable from both environmental and economic standpoints. In this review, we summarized the vitamins which can potentially be produced using microbial cell factories or are already being produced in commercial fermentation processes. They include water-soluble vitamins (vitamin B complex and vitamin C) as well as fat-soluble vitamins (vitamin A/D/E and vitamin K). Furthermore, metabolic engineering is discussed to provide a reference for the construction of microbial cell factories. We also highlight the current state and problems encountered in the fermentative production of vitamins.

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A supernumerary designer chromosome for modular in vivo pathway assembly in Saccharomyces cerevisiae

10.1101/2020.02.18.954131 ◽

2020 ◽

Cited By ~ 1

Author(s):

Eline D. Postma ◽

Sofia Dashko ◽

Lars van Breemen ◽

Shannara K. Taylor Parkins ◽

Marcel van den Broek ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Genome Engineering ◽

De Novo ◽

Transcriptional Unit ◽

Dna Assembly ◽

Cellular Processes ◽

Microbial Cell Factories ◽

Cell Factories ◽

Expression Platform

ABSTRACTThe construction of microbial cell factories for sustainable production of chemicals and pharmaceuticals requires extensive genome engineering. Using Saccharomyces cerevisiae, this study proposes Synthetic Chromosomes (SynChs) as orthogonal expression platforms for rewiring native cellular processes and implementing new functionalities. Capitalizing the powerful homologous recombination capability of S. cerevisiae, modular SynChs of 50 and 100 Kb were fully assembled de novo from up to 44 transcriptional-unit-sized fragments in a single transformation. These assemblies were remarkably efficient and faithful to their in silico design. SynChs made of non-coding DNA were stably replicated and segregated irrespective of their size without affecting the physiology of their host. These non-coding SynChs were successfully used as landing pad and as exclusive expression platform for the essential glycolytic pathway. This work pushes the limit of DNA assembly in S. cerevisiae and paves the way for de novo designer chromosomes as modular genome engineering platforms in S. cerevisiae.

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Synthetic Biology Approaches to Engineer Saccharomyces cerevisiae towards the Industrial Production of Valuable Polyphenolic Compounds

Life ◽

10.3390/life10050056 ◽

2020 ◽

Vol 10 (5) ◽

pp. 56 ◽

Cited By ~ 2

Author(s):

João Rainha ◽

Daniela Gomes ◽

Lígia R. Rodrigues ◽

Joana L. Rodrigues

Keyword(s):

Saccharomyces Cerevisiae ◽

Synthetic Biology ◽

Plant Secondary Metabolites ◽

Genetically Engineered ◽

Microbial Cell ◽

Bioactive Molecules ◽

Microbial Cell Factories ◽

Cell Factories ◽

Increasing Demand ◽

Engineered Saccharomyces Cerevisiae

Polyphenols are plant secondary metabolites with diverse biological and potential therapeutic activities such as antioxidant, anti-inflammatory and anticancer, among others. However, their extraction from the native plants is not enough to satisfy the increasing demand for this type of compounds. The development of microbial cell factories to effectively produce polyphenols may represent the most attractive solution to overcome this limitation and produce high amounts of these bioactive molecules. With the advances in the synthetic biology field, the development of efficient microbial cell factories has become easier, largely due to the development of the molecular biology techniques and by the identification of novel isoenzymes in plants or simpler organisms to construct the heterologous pathways. Furthermore, efforts have been made to make the process more profitable through improvements in the host chassis. In this review, advances in the production of polyphenols by genetically engineered Saccharomyces cerevisiae as well as by synthetic biology and metabolic engineering approaches to improve the production of these compounds at industrial settings are discussed.

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Metabolic engineering of Saccharomyces cerevisiae microbial cell factories for succinic acid production

Journal of Biotechnology ◽

10.1016/j.jbiotec.2007.07.367 ◽

2007 ◽

Vol 131 (2) ◽

pp. S205 ◽

Cited By ~ 7

Author(s):

Jose Manuel Otero ◽

Lisbeth Olsson ◽

Jens Nielsen

Keyword(s):

Saccharomyces Cerevisiae ◽

Metabolic Engineering ◽

Succinic Acid ◽

Acid Production ◽

Microbial Cell ◽

Microbial Cell Factories ◽

Cell Factories ◽

Succinic Acid Production

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The Plasma Membrane at the Cornerstone Between Flexibility and Adaptability: Implications for Saccharomyces cerevisiae as a Cell Factory

Frontiers in Microbiology ◽

10.3389/fmicb.2021.715891 ◽

2021 ◽

Vol 12 ◽

Author(s):

Luís Ferraz ◽

Michael Sauer ◽

Maria João Sousa ◽

Paola Branduardi

Keyword(s):

Saccharomyces Cerevisiae ◽

Plasma Membrane ◽

Cell Factory ◽

Membrane Composition ◽

Microbial Cell Factories ◽

Cell Factories ◽

A Cell ◽

Biotechnological Processes ◽

Membrane Engineering ◽

The Impact

In the last decade, microbial-based biotechnological processes are paving the way toward sustainability as they implemented the use of renewable feedstocks. Nonetheless, the viability and competitiveness of these processes are often limited due to harsh conditions such as: the presence of feedstock-derived inhibitors including weak acids, non-uniform nature of the substrates, osmotic pressure, high temperature, extreme pH. These factors are detrimental for microbial cell factories as a whole, but more specifically the impact on the cell’s membrane is often overlooked. The plasma membrane is a complex system involved in major biological processes, including establishing and maintaining transmembrane gradients, controlling uptake and secretion, intercellular and intracellular communication, cell to cell recognition and cell’s physical protection. Therefore, when designing strategies for the development of versatile, robust and efficient cell factories ready to tackle the harshness of industrial processes while delivering high values of yield, titer and productivity, the plasma membrane has to be considered. Plasma membrane composition comprises diverse macromolecules and it is not constant, as cells adapt it according to the surrounding environment. Remarkably, membrane-specific traits are emerging properties of the system and therefore it is not trivial to predict which membrane composition is advantageous under certain conditions. This review includes an overview of membrane engineering strategies applied to Saccharomyces cerevisiae to enhance its fitness under industrially relevant conditions as well as strategies to increase microbial production of the metabolites of interest.

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Od rastlinske biomase do biogoriv in bio-surovin z mikrobnimi celičnimi tovarnami

Acta agriculturae Slovenica ◽

10.14720/aas.2020.116.1.1331 ◽

2020 ◽

Vol 116 (1) ◽

pp. 75

Author(s):

Maša VODOVNIK ◽

Matevž ZLATNAR

Keyword(s):

Renewable Energy ◽

Plant Biomass ◽

Consolidated Bioprocessing ◽

Raw Materials ◽

Value Added ◽

Cost Effective ◽

Human Society ◽

Microbial Cell Factories ◽

Cell Factories ◽

Wide Range

<p>Global energy demands and global warming represent key challenges of the future of human society. Continous renewable energy supply is key for sustainable economy development. Waste plant biomass represent abundant source of renewable energy that can be transformed to biofuels and other value-added products, which is currently limited due to the lack of cost-effective biocatalysts. The bottleneck of this process is the degradation of structural polysaccharides of plant cell walls to soluble compounds that can be fermented to solvents or transformed to biogas via methanogenesis and can be used as biofuels or chemical raw materials. In order to replace traditional physical and chemical methods of lignocellulose pretreatment with more environmentally friendly biological approaches, native microbial enzyme systems are increasingly being explored as potential biocatalysts that could be used in these processes. Microbial enzymes are useful either as catalysts in the enzymatic hydrolysis of lignocelluloses or as components incorporated in engineered microbes for consolidated bioprocessing of lignocelluloses. The unprecedented development of tools for genetic and metabolic engineering for a wide range of microorganisms enabled significant progress in the development of microbial cell factories optimized for the producton of biofuels. One of the most promising strategies aimed towards this goal, i.e. systematic design and heterologous expression of »designer cellulosomes« in industrial solventogenic strains is adressed in detail.</p>

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Enzymatic Hydrolysate of Cinnamon Waste Material as Feedstock for the Microbial Production of Carotenoids

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18031146 ◽

2021 ◽

Vol 18 (3) ◽

pp. 1146

Author(s):

Stefano Bertacchi ◽

Stefania Pagliari ◽

Chiara Cantù ◽

Ilaria Bruni ◽

Massimo Labra ◽

...

Keyword(s):

Industrial Sector ◽

Waste Material ◽

Fungal Cell ◽

Enzymatic Hydrolysate ◽

Carbon And Nitrogen ◽

Microbial Cell Factories ◽

Cell Factories ◽

Separate Hydrolysis And Fermentation ◽

Additional Processing ◽

Microbial Cultivation

In the context of the global need to move towards circular economies, microbial cell factories can be employed thanks to their ability to use side-stream biomasses from the agro-industrial sector to obtain additional products. The valorization of residues allows for better and complete use of natural resources and, at the same time, for the avoidance of waste management to address our needs. In this work, we focused our attention on the microbial valorization of cinnamon waste material after polyphenol extraction (C-PEW) (Cinnamomum verum J.Presl), generally discarded without any additional processing. The sugars embedded in C-PEW were released by enzymatic hydrolysis, more compatible than acid hydrolysis with the subsequent microbial cultivation. We demonstrated that the yeast Rhodosporidium toruloides was able to grow and produce up to 2.00 (±0.23) mg/L of carotenoids in the resulting hydrolysate as a sole carbon and nitrogen source despite the presence of antimicrobial compounds typical of cinnamon. To further extend the potential of our finding, we tested other fungal cell factories for growth on the same media. Overall, these results are opening the possibility to develop separate hydrolysis and fermentation (SHF) bioprocesses based on C-PEW and microbial biotransformation to obtain high-value molecules.

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Protein-Engineered Polymers Functionalized with Antimicrobial Peptides for the Development of Active Surfaces

Applied Sciences ◽

10.3390/app11125352 ◽

2021 ◽

Vol 11 (12) ◽

pp. 5352

Author(s):

Ana Margarida Pereira ◽

Diana Gomes ◽

André da Costa ◽

Simoni Campos Dias ◽

Margarida Casal ◽

...

Keyword(s):

Antimicrobial Activity ◽

Antimicrobial Peptides ◽

Recombinant Dna ◽

Biological Properties ◽

Resistant Bacteria ◽

Recombinant Dna Technology ◽

Free Standing ◽

Microbial Cell Factories ◽

Cell Factories ◽

Antimicrobial Materials

Antibacterial resistance is a major worldwide threat due to the increasing number of infections caused by antibiotic-resistant bacteria with medical devices being a major source of these infections. This suggests the need for new antimicrobial biomaterial designs able to withstand the increasing pressure of antimicrobial resistance. Recombinant protein polymers (rPPs) are an emerging class of nature-inspired biopolymers with unique chemical, physical and biological properties. These polymers can be functionalized with antimicrobial molecules utilizing recombinant DNA technology and then produced in microbial cell factories. In this work, we report the functionalization of rPBPs based on elastin and silk-elastin with different antimicrobial peptides (AMPs). These polymers were produced in Escherichia coli, successfully purified by employing non-chromatographic processes, and used for the production of free-standing films. The antimicrobial activity of the materials was evaluated against Gram-positive and Gram-negative bacteria, and results showed that the polymers demonstrated antimicrobial activity, pointing out the potential of these biopolymers for the development of new advanced antimicrobial materials.

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