To achieve equilibrium among the three modules, we implemented promoter engineering, culminating in the development of an engineered E. coli TRP9 strain. A 5-liter fermentor, subjected to fed-batch cultivation, produced a tryptophan titer of 3608 g/L, signifying a yield of 1855%, which constitutes 817% of the theoretically highest attainable yield. The strain that produces tryptophan with a high yield provided a solid basis for the large-scale manufacturing of tryptophan.
In the context of synthetic biology, Saccharomyces cerevisiae, a microorganism generally acknowledged as safe, is a extensively studied chassis cell for the production of high-value or bulk chemicals. Metabolic engineering techniques have led to the development and optimization of a significant number of chemical synthesis pathways in S. cerevisiae, and the consequent production of specific chemicals presents a path to commercialization. S. cerevisiae, being a eukaryote, has a complete internal membrane system and intricate organelle compartments. These compartments frequently hold elevated levels of precursor substrates such as acetyl-CoA in mitochondria, or contain sufficient enzymes, cofactors, and energy for the synthesis of certain chemicals. These attributes might create a more suitable physical and chemical environment, thereby supporting the biosynthesis of the target chemicals. In contrast, the structural variations in different organelles are detrimental to the synthesis of particular chemicals. Researchers have refined the process of product biosynthesis by meticulously altering organelles. This refinement process has been guided by an in-depth analysis of organelle properties and the alignment of target chemical biosynthesis pathways with the characteristics of individual organelles. This review thoroughly examines the reconstruction and optimization of chemical biosynthesis pathways in Saccharomyces cerevisiae, specifically focusing on the organelles: mitochondria, peroxisomes, Golgi apparatus, endoplasmic reticulum, lipid droplets, and vacuoles. Current difficulties, challenges, and future perspectives are emphasized.
A non-conventional red yeast, Rhodotorula toruloides, possesses the capability of synthesizing a multitude of carotenoids and lipids. Various inexpensive raw materials can be employed, and this process can also withstand and absorb harmful compounds in lignocellulosic hydrolysate. Currently, research extensively focuses on the production of microbial lipids, terpenes, high-value enzymes, sugar alcohols, and polyketides. Researchers have conducted extensive theoretical and technological exploration across genomics, transcriptomics, proteomics, and a genetic operation platform, driven by the perceived broad industrial application opportunities. We scrutinize the recent progress in *R. toruloides*' metabolic engineering and natural product synthesis, and then explore the future challenges and potential solutions for developing a *R. toruloides* cell factory.
Non-conventional yeasts, including Yarrowia lipolytica, Pichia pastoris, Kluyveromyces marxianus, Rhodosporidium toruloides, and Hansenula polymorpha, are demonstrated as effective cell factories in producing diverse natural products due to their wide adaptability to various substrates, significant resilience to harsh environmental factors, and other remarkable characteristics. Fueled by the progress in synthetic biology and gene editing, metabolic engineering techniques for non-conventional yeasts are undergoing a period of considerable growth and diversification. Wnt inhibitor This review delves into the physiological aspects, tool design and present-day usage of multiple prominent non-conventional yeast strains, followed by a compilation of common metabolic engineering methodologies used to enhance natural product biosynthesis. We delve into the capabilities and limitations of using non-conventional yeasts as natural product cell factories in the current context, and outline promising future research and development avenues.
Plant-derived diterpenoids, a diverse class of compounds, showcase a wide range of structural forms and functions. In the pharmaceutical, cosmetic, and food additive industries, these compounds are widely employed due to their pharmacological characteristics, including anticancer, anti-inflammatory, and antibacterial properties. In the recent years, the identification of functional genes within plant-derived diterpenoid biosynthetic pathways has progressed alongside the advancements in synthetic biotechnology. This has spurred considerable efforts in developing varied microbial cell factories for diterpenoids via metabolic engineering and synthetic biology. The outcome has been a gram-level production of a wide spectrum of these compounds. Synthetic biology is employed in this article to detail the construction of microbial cell factories that produce plant-derived diterpenoids. Subsequently, it elucidates metabolic engineering strategies used to increase diterpenoid production, with the objective of offering a guide for establishing high-yielding systems for industrial production.
Throughout living organisms, S-adenosyl-l-methionine (SAM) is consistently present and plays a significant part in transmethylation, transsulfuration, and transamination. SAM production, due to its vital physiological functions, has experienced a surge in attention. In current SAM production research, microbial fermentation is the primary method of choice. This method is significantly more cost-effective than chemical synthesis or enzyme catalysis, making commercial production more straightforward. With the remarkable growth in the demand for SAM, there was an increase in the pursuit of creating microorganisms that produced exceptionally high amounts of SAM. Microorganisms' SAM productivity can be elevated through the combined efforts of conventional breeding and metabolic engineering. A review of recent research efforts to elevate microbial S-adenosylmethionine (SAM) production is presented, highlighting the potential to advance overall SAM productivity. Furthermore, the study included a discussion of the roadblocks in SAM biosynthesis and their potential solutions.
Organic compounds, specifically organic acids, are formed through the use of biological systems for their synthesis. These substances frequently include one or more low molecular weight acidic groups, like carboxyl and sulphonic groups. Food, agriculture, medicine, bio-based materials, and other sectors all heavily rely on organic acids for their various purposes. Yeast's benefits encompass unparalleled biosafety, strong stress resistance across various conditions, a diverse spectrum of utilizable substrates, convenient genetic manipulation, and a well-established large-scale cultivation procedure. For this reason, the application of yeast to generate organic acids is compelling. Medicines information Still, challenges involving low concentration, an abundance of by-products, and an inefficient fermentation process continue. The application of yeast metabolic engineering and synthetic biology techniques has yielded considerable progress in this field recently. Here, we provide a summary of the progress in yeast's production of 11 organic acids. These organic acids are composed of bulk carboxylic acids and high-value organic acids, both of which may be produced naturally or heterologously. Finally, the potential of this field in the future was articulated.
Polyisoprenoids and scaffold proteins make up functional membrane microdomains (FMMs), which are integral to diverse cellular physiological processes found in bacteria. A key objective of this study was to identify the correlation between MK-7 and FMMs, with the subsequent aim of controlling MK-7 biosynthesis through the use of FMMs. Utilizing fluorescent labeling, the study determined the interplay between FMMs and MK-7 within the cellular membrane. Subsequently, an analysis of MK-7's role as a crucial polyisoprenoid component within FMMs involved observing modifications in MK-7 membrane content and membrane order before and after disrupting the integrity of FMMs. Using visual techniques, the subcellular location of critical MK-7 synthesis enzymes was determined. The intracellular free enzymes, Fni, IspA, HepT, and YuxO, were found localized in FMMs, achieved by the protein FloA, which led to the compartmentalization of the MK-7 synthetic pathway. Eventually, a high MK-7 production strain—BS3AT—was successfully obtained. In comparison to the 3003 mg/L production in shake flasks, the 3-liter fermenter achieved a significantly higher production rate of 4642 mg/L for MK-7.
Natural skin care products often find a valuable ingredient in tetraacetyl phytosphingosine (TAPS). Deacetylation of the substance yields phytosphingosine, a key component for creating ceramide, a moisturizing ingredient in skincare products. Accordingly, TAPS holds a prominent position in the skincare-oriented cosmetic industry. The yeast Wickerhamomyces ciferrii, a non-standard microbe, is uniquely recognized for naturally secreting TAPS, thus positioning it as the sole host for industrial TAPS production. immunity heterogeneity First, this review introduces the discovery and functions of TAPS. Subsequently, the metabolic pathway for its biosynthesis is described in detail. The subsequent strategies for enhancing TAPS production in W. ciferrii are outlined, incorporating haploid screening, mutagenesis breeding, and metabolic engineering approaches. Moreover, the possibilities for TAPS biomanufacturing using W. ciferrii are considered, taking into account the current developments, difficulties, and trends in the field. Ultimately, a blueprint for engineering W. ciferrii cell factories, leveraging synthetic biology principles, to produce TAPS is also provided.
Essential for the balanced hormonal system within a plant and for regulating both growth and metabolism, abscisic acid is a plant hormone that hinders growth. From crop improvement to medical advancements, abscisic acid's versatile properties, including its effect on drought and salt tolerance, reduction of fruit browning, mitigation of malaria transmission, and promotion of insulin production, are valuable in agricultural and medicinal contexts.