Biotechnology Notes最新文献

Furthering the development of the field of synthetic biology in Thailand is included in the Thai government's Bio-Circular-Green (BCG) economic policy. The BCG model has increased collaborations between government, academia and private sectors with the specific aim of increasing the value of bioindustries via sustainable approaches. This article provides a critical review of current academic research related to synthetic biology conducted in Thailand during the last decade including genetic manipulation, metabolic engineering, cofactor enhancement to produce valuable chemicals, and analysis of synthetic cells using systems biology. Work was grouped according to a Design-Build-Test-Learn cycle. Technical areas directly supporting development of synthetic biology for BCG in the future such as enzyme catalysis, enzyme engineering and systems biology related to culture conditions are also discussed. Key activities towards development of synthetic biology in Thailand are also discussed.

Reprogrammed glucose-responsive, insulin + cells (“β-like”) exhibit the potential to bypass the hurdles of exogenous insulin delivery in treating diabetes mellitus. Current cell-based therapies-transcription factor regulation, biomolecule-mediated enteric signaling, and transgenics - have demonstrated the promise of reprogramming either mature or progenitor gut cells into surrogate “β-like” cells. However, there are predominant challenges impeding the use of gut “β-like” cells as clinical replacements for insulin therapy. Reprogrammed “β-like” gut cells, even those of enteroendocrine origin, mostly do not exhibit glucose – potentiated insulin secretion. Despite the exceptionally low conversion rate of gut cells into surrogate “β-like” cells, the therapeutic quantity of gut “β-like” cells needed for normoglycemia has not even been established. There is also a lingering uncertainty regarding the functionality and bioavailability of gut derived insulin. Herein, we review the strategies, challenges, and opportunities in the generation of functional, reprogrammed “β-like” cells.

The bio-manufacturing of products with substantial commercial value, particularly polyhydroxyalkanoates (PHA), using cost-effective carbon sources through microorganisms, has garnered heightened attention from both the scientific community and industry over the past few decades. Opting for industrial PHA production from various organic wastes, spanning industrial, agricultural, municipal, and food-based sources, emerges as a wiser choice. This strategy not only eases the burden of recycling organic waste and curbs environmental pollution but also trims down PHA production costs, rendering these materials more competitive in commercial markets. In addition, PHAs are a family of renewable, environmentally friendly, fully biodegradable and biocompatible polyesters with a multitude of applications. This review provides an overview of recent developments in PHA production from organic wastes. It covers the optimization of diverse metabolic pathways for producing various types of PHA from organic waste sources, pre-treatment and downstream processing for PHA using unrelated organic wastes, and challenges in industrial production of PHA using unrelated organic waste feedstocks and the challenges faced in industrial PHA production from organic wastes, along with potential solutions. Lastly, this study suggests underlying research endeavors aimed at further enhancing of the feasibility of industrial PHA production from organic wastes as an alternative to current petroleum-based plastics in the near future.

Insect olfaction directly impacts insect behavior and thus is an important consideration in the development of smart farming tools and in integrated pest management strategies. Insect olfactory receptors (ORs) have been traditionally studied using Drosophila empty neuron systems or with expression and functionalization in HEK293 cells or Xenopus laevis oocytes. Recently, the yeast Saccharomyces cerevisiae (S. cerevisiae) has emerged as a promising chassis for the functional expression of heterologous seven transmembrane receptors. S. cerevisiae provides a platform for the cheap and high throughput study of these receptors and potential deorphanization. In this study, we explore the foundations of a scalable yeast-based platform for the functional expression of insect olfactory receptors by employing a genetically encoded calcium sensor for quantitative evaluation of fluorescence and optimized experimental parameters for enhanced functionality. While the co-receptor of insect olfactory receptors remains non-functional in our yeast-based system, we thoroughly evaluated various experimental variables and identified future research directions for establishing an OR platform in S. cerevisiae.

SynBio Africa is a forum for researchers, students, citizen scientists, policymakers and the public to convene and develop successful pathways for the propagation of synthetic biology technologies, products, and services throughout Africa. Our vision is to have a healthy, safe, and sustainable world through synthetic biology. In Africa, synthetic biology has the potential to greatly contribute to national development agenda through the following ways: i) by anchoring a sustainable bioeconomy; ii) by helping develop innovative medicines; iii) by reducing pollution, and iv) by increasing crop production to reduce hunger. However, there is little to no information on synthetic biology and its regulatory policies in Africa. Across the continent, scientists, policy makers, researchers and others are still working in silos—only partaking in consultative meetings to try and develop a set of unified policy guidelines. SynBio Africa is therefore proposing to establish the first Center of Excellence in Synthetic Biology in Africa with six themes, namely: research, capacity development, innovation hub, biosafety and biosecurity, and bioinformatics and data science, and one-health. Accordingly, SynBio Africa will work with collaborators from government and non-governmental organizations, the public and private sectors, and educational institutions from Uganda, Africa, and around the world to implement these six themes.

Yarrowia lipolytica is a modern workhorse for biotechnology that is amenable to genetic manipulations and can produce high levels of various enzymes. The present study was designed to engineer Y. lipolytica for the overexpression of α-bisabolene, a valuable biofuel precursor and pharmaceutical, making use of this yeast's ability to accumulate lipids, and with the use of a golden gate DNA assembly (GG) toolbox. By transforming Y. lipolytica with a GG genetic construct involving truncated 3-hydroxy-3-methyl-glutaryle coenzyme A reductase (tHMG) and α-bisabolene synthase (Bis) genes controlled by the strong TEF promoter and Lip2 terminator, the engineered yeast was able to produce 489 mg l⁻¹ of α-bisabolene. This was increased to 816 mg l⁻¹ by transforming a lipid-over-accumulating Y. lipolytica strain with the same genetic construct. Higher production titers of up to 1243 mg l⁻¹ could be also achieved by varying the culture conditions of the transformed strains.

Fe doped MgO nanoparticles were synthesized using a straightforward soft chemical method. We conducted a comprehensive examination of the electrical properties of Fe-doped MgO nanoparticles with a crystalline size range of 7–10 nm. Simultaneously, we explored their antibacterial capabilities. Our findings indicate that an increase in the concentration of Fe-doped MgO correlates with an enhanced bactericidal effect. To gain a deeper understanding of charge transfer processes, the AC conductivity and dielectric characteristics of the samples across various temperatures and frequencies was studied.The antibacterial activity was studied utilising the MIC methodology, the live count (LC) method, and the agar cup technique in addition to the electrical characteristics. After exposure to nanoparticles, we observed the disruption of pathogenic cell walls through transmission electron microscopy (TEM) analysis. These results suggest that Fe-doped MgO nanoparticles hold promise for the development of novel, more effective antibacterial drugs. The ½ MIC for E.coli was found to be 2.75 μg/ml, while for Bacillus sp., it was 1.75 μg/ml when exposed to Fe-doped MgO nanoparticles. This dosage level may find applications in the medical field. However, further investigations are required to assess potential toxicity and long-term environmental and human health effects. If successful in vivo tests follow, Fe-doped MgO nanoparticles could emerge as valuable antibacterial agents.

Enhanced synthesis of hyaluronic acid (HA) with recombinant Corynebacterium glutamicum as production host was achieved in this work. Hyaluronan synthase (HAS), which is a membrane protein acting as a key enzyme in HA biosynthesis, impacts both HA yield and its molecular weight. Cell morphology, which includes size, shape, and surface area, has a large impact on the expression and activity of HAS. Therefore, deliberate regulation of cell morphology holds the potential to enhance HA production. Here, we constructed three modules, namely the transporter module, the morphology tuning module and the HA synthesis module. The transporter module contains a strong constitutive promoter P_tuf and arabinose transport protein was used to control the maximum amount of inducer entering the cell, thus reducing excessive cell deformation. The morphology tuning module contains an arabinose-inducible weak promoter P_BAD and a cell-division-relevant gene was used to sense intracellular inducer concentrations and achieve different degrees of change in cell size. These two modules worked together, described as a dual-valve regulation, to achieve fine-tuning of cell morphology, resulting in a 1.87-fold increase in cell length and a 2.08-fold increase in cell membrane. When combined with the HA synthesis module, the HA titer reached 16.0 g/L, which was 1.6 times the yield reported in the previous morphology-engineered strain. Hence, for the first time, a morphologically engineered strain resulting in both high cell density and HA titer was constructed.

Circular mRNA (circmRNA) is a covalent closed loop formed by reverse splicing of the 3′ end to the 5′ end of mRNA. Compared to traditional linear mRNAs, circmRNAs can mediate efficient, stable, and durable protein expression and are considered an alternative to linear mRNAs in terms of therapeutic reagents. With the continuous development of circmRNA research, circmRNA has also made significant progress in vaccines and cellular therapies. In this review, we present research advances in the in vitro synthesis of circmRNAs, focusing on the biological ligation methods of circmRNAs and current applications, with a summary of challenges regarding circmRNA design, synthesis, and applications. Based on the enhanced stability of circmRNAs, further research on circmRNAs could help expand their applications in biotherapeutics and strengthen their role in basic medical applications.