Engineering Microbiology最新文献

Cyclodipeptides are diverse chemical scaffolds that show a broad range of bioactivities relevant for medicine, agriculture, chemical catalysis, and material sciences. Cyclodipeptides can be synthesized enzymatically through two unrelated enzyme families, non-ribosomal peptide synthetases (NRPS) and cyclodipeptide synthases (CDPSs). The chemical diversity of cyclodipeptides is derived from the two amino acid side chains and the modification of those side-chains by cyclodipeptide tailoring enzymes. While a large spectrum of chemical diversity is already known today, additional chemical space - and as such potential new bioactivities - could be accessed by exploring yet undiscovered NRPS and CDPS gene clusters as well as via engineering. Further, to exploit cyclodipeptides for applications, the low yield of natural biosynthesis needs to be overcome. In this review we summarize current knowledge on NRPS and CDPS-based cyclodipeptide biosynthesis, engineering approaches to further diversity the natural chemical diversity as well as strategies for high-yield production of cyclodipeptides, including a discussion of how advancements in synthetic biology and metabolic engineering can accelerate the translational potential of cyclodipeptides.

In recent decades, third-generation (3G) biofuels have become a more attractive method of fuel production, as algae cultivation does not infringe on resources needed for food production. Additionally, algae can adapt to different environments, has high photosynthetic efficiency (CO₂ fixation), and has a high potential for carbohydrate accumulation. The prevalence of algae worldwide demonstrates its ability to adapt to different environments and climates, proving its biodiversity and versatility. Algae can be grown in wastewater, seawater, and even sewage, thus ensuring a lower water footprint and greater energy efficiency during algal biomass production. Because of this, the optimization of 3G ethanol production appears to be an excellent alternative to mitigate environmental impacts and increase energy and food security. This critical review presents (i) the stages of cultivation and processing of micro and macroalgae; (ii) the selection of yeasts (through engineering and/or bioprospecting) to produce ethanol from these biomasses; (iii) the potential of seawater-based facilities to reduce water footprint; and (iv) the mass and energy balances of 3G ethanol production in the world energy matrix. This article is, above all, a brainstorm on the environmental viability of algae bioethanol.

Predicting biodiversity and dynamics of complex communities is a fundamental challenge in ecology. Leveraging bacterial microcosms with well-controlled laboratory conditions, Hu et al. recently performed a direct test of theory predicting that two community-level parameters (i.e., species pool size and inter-species interaction strength) dictate transitions between three dynamical phases: stable full coexistence, stable partial coexistence, and persistent fluctuations. Generally, communities experience species extinctions before they lose stability as either of the two parameters increases.

Diterpenes, or diterpenoids, are the most abundant and diverse subgroup of terpenoids, the largest family of secondary metabolites. Most diterpenes possess broad biological activities including anti-inflammatory, antiviral, anti-tumoral, antimicrobial, anticancer, antifungal, antidiabetic, cardiovascular protective, and phytohormone activities. As such, diterpenes have wide applications in medicine (e.g., the anticancer drug Taxol and the antibiotic pleuromutilin), agriculture (especially as phytohormones such as gibberellins), personal care (e.g., the fragrance sclareol) and food (e.g., steviol glucosides as low-calorie sweeteners) industries. Diterpenes are biosynthesized in a common route with various diterpene synthases and decoration enzymes like cytochrome P450 oxidases, glycosidases, and acyltransferases. Recent advances in DNA sequencing and synthesis, omics analysis, synthetic biology, and metabolic engineering have enabled efficient production of diterpenes in several chassis hosts like Escherichia coli, Saccharomyces cerevisiae, Yarrowia lipolytica, Rhodosporidium toruloides, and Fusarium fujikuroi. This review summarizes the recently discovered diterpenes, their related enzymes and biosynthetic pathways, particularly highlighting the microbial synthesis of high-value diterpenes directly from inexpensive carbon sources (e.g., sugars). The high titers (>4 g/L) achieved mean that some of these endeavors are reaching or close to commercialization. As such, we envisage a bright future in translating microbial synthesis of diterpenes into commercialization.

The microbiome is an essential component of ecological systems and is comprised of a diverse array of microbes. Over the past decades, the accumulated observational evidence reveals a close correlation between the microbiome and human health and disease. Many groups are now manipulating individual microbial strains, species and the community as a whole to gain a mechanistic understanding of the functions of the microbiome. Here, we discuss three major approaches for introducing DNA to engineer model bacteria and isolated undomesticated bacteria, including transformation, transduction, and conjugation. We provide an overview of these approaches and describe the advantages and limitations of each method. In addition, we highlight examples of human microbiome engineering using these approaches. Finally, we provide perspectives for the future of microbiome engineering.

The production of biofuels and biochemicals derived from microbial fermentation has received a lot of attention and interest in light of concerns about the depletion of fossil fuel resources and climatic degeneration. However, the economic viability of feedstocks for biological conversion remains a barrier, urging researchers to develop renewable and sustainable low-cost carbon sources for future bioindustries. Owing to the numerous advantages, acetate has been regarded as a promising feedstock targeting the production of acetyl-CoA-derived chemicals. This review aims to highlight the potential of acetate as a building block in industrial biotechnology for the production of bio-based chemicals with metabolic engineering. Different alternative approaches and routes comprised of lignocellulosic biomass, waste streams, and C1 gas for acetate generation are briefly described and evaluated. Then, a thorough explanation of the metabolic pathway for biotechnological acetate conversion, cellular transport, and toxin tolerance is described. Particularly, current developments in metabolic engineering of the manufacture of biochemicals from acetate are summarized in detail, with various microbial cell factories and strategies proposed to improve acetate assimilation and enhance product formation. Challenges and future development for acetate generation and assimilation as well as chemicals production from acetate is eventually shown. This review provides an overview of the current status of acetate utilization and proves the great potential of acetate with metabolic engineering in industrial biotechnology.

Lactate is an important bulk chemical with widespread applications and a major byproduct of other chemicals bioprocess in microbial fermentation. Lactate dehydrogenase A (LdhA) catalyzes the synthesis of lactate from pyruvate. Lysine acetylation is an evolutionarily conserved post-translational modification; however, the mechanisms underlying the regulation of LdhA function by lysine acetylation in Escherichia coli remain poorly understood. Herein, we demonstrate acetylation of E. coli LdhA occurs via enzymatic and non-enzymatic mechanisms. Further, we show carbon source type and concentration affect the lysine acetylation status of LdhA via a non-enzymatic mechanism. Lysine acetylation significantly inhibits the enzymatic activity and protein level of LdhA. The results of the present study demonstrate lysine acetylation of E. coli LdhA is irreversible. Understanding of the effects of lysine acetylation on LdhA function may provide a new perspective for regulating lactate production in microbial synthesis.

The de novo biosynthesis of vindoline and catharanthine, the direct precursors used for industrial production of the anti-cancer drug vinblastine, has been achieved in the yeast cell factory. To date, this is the longest natural product biosynthesis pathway that has been successfully transferred from plants to microorganisms, indicating the possibility of producing more than 3,000 other monoterpene indole alkaloids in yeast by synthetic genome engineering.

Phage-encoded homologous recombination (PEHR) is an efficient tool for bacterial genome editing. We previously developed and utilized a Pseudomonas-specific PEHR system. However, when using the PEHR system for Pseudomonas genome editing, false positives can be a problem. In this study, we combined a compact Cascade-Cas3 system from P. aeruginosa (PaeCas3c) with a Pseudomonas-specific PEHR system, and the results of our recombineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.

Secondary metabolites in microorganisms represent a resource for drug discovery and development. In particular, microbial-derived antitumor agents are in clinical use worldwide. Herein, we provide an overview of the development of classical antitumor drugs derived from microorganisms. Currently used drugs and drug candidates are comprehensively described in terms of pharmacological activities, mechanisms of action, microbial sources, and biosynthesis. We further discuss recent studies that have demonstrated the utility of gene-editing technologies and synthetic biology tools for the identification of new gene clusters, expansion of natural products, and elucidation of biosynthetic pathways. This review summarizes recent progress in the discovery and development of microbial-derived anticancer compounds with emphasis on biosynthesis.