Biotechnology Notes最新文献

Synthetic biology has gained many interest around the globe in the last two decades, not only due to its rapid development but also the potential to provide addressable solutions using standardized design of biological systems. Considering its huge population, biodiversity, and natural resources, Indonesia could play an important role in shaping the future of synthetic biology towards a sustainable bio-circular economy. Here, we provide an overview of synthetic biology development in Indonesia, especially on exploring the potential of our biodiversity. We also discuss some potentials of synthetic biology in solving national issues. Furthermore, we also provide the projection and future landscape of synthetic biology development in Indonesia. In addition, we briefly explain the potential challenges that may arise during the development.

The extensive use of chemical dyes, primarily Azo and anthraquinone dyes, in textiles has resulted in their alarming release into the environment by textile industries. The introduction of heavy metals into these dyes leads to an increase in Biochemical Oxygen Demand (BOD), Chemical Oxygen Demand (COD), and water toxicity. Conventional physicochemical methods for treating textile effluents are costly and energy-intensive. Here introduction of new strategies is eminent, microbial bioremediation for the biodegradation and detoxification of these hazardous dyes, possesses as an innovative solution for the existing problem, discussed are specific groups of bacteria, fungi, and algae which could be one of the potential decolorizing agents that could replace the majority of other expensive processes in textile wastewater treatment by using enzymes like peroxidase, laccase, and azoreductase. These enzymes catalyzes chemical reactions that break down the dye molecules into less harmful substances. Additionally, novel strategies and advancements to enhance the effectiveness of these microbes and their products are comprehensively discussed.

Due to its abundance, cost-effectiveness, and high reducibility, methanol has gained considerable attention in the biomanufacturing industry as a nonfood feedstock for the production of value-added chemicals. The range of chemicals that can be derived from methanol, however, remains constrained and is currently in the concept validation phase. This study aimed to develop and evaluate a hybrid methanol assimilation pathway in Escherichia coli to improve the production of (R)-1,3-butanediol ((R)-1,3-BDO) by utilizing methanol and sugars as co-substrates. By combining the methanol dehydrogenase (MDH) from the prokaryotes with the dihydroxyacetone synthase (DAS) from the eukaryotes, the hybrid pathway facilitates methanol conversion into the central metabolism while generating NADH at the same time. Through pathway optimization and targeted gene deletions, we have successfully developed an E. coli strain capable of producing 5.79 g/L (R)-1,3-BDO in shake flask experiments and 13.71 g/L (R)-1,3-BDO with a yield of 0.35 C-mol/C-mol in batch fermentation using methanol and glucose as co-substrates. Our study also showed the incorporation of ¹³C-methanol into cellular intermediates and an increase in NAD(P)H concentration, confirming the role of methanol as a co-substrate and supplier of NADH. In addition, our study also demonstrated the co-utilization of methanol with xylose for the production of (R)-1,3-BDO, expanding the substrate spectrum for sustainable 1,3-BDO production.

Plasmids are one of the most commonly used basic tools in the construction of microbial cell factories, the use of which individually or in pairs play an important role in the expression of exogenous gene modules. However, little attention has been paid to the interactions of plasmid-plasmid and plasmid-host in the widespread use of the double plasmid system. In this study, we demonstrated that dual-plasmid interactions facilitated to cell growth and product accumulation in Saccharomyces cerevisiae. The strain containing both the expression plasmid pEV (a plasmid carrying the gene encoding valencene synthase) and the assistant plasmid pI (an empty plasmid expressing no extra gene) showed a significant improvement in relative growth rate, biomass and valencene production compared to the strain containing only the pEV plasmid. The transcriptional level analysis revealed an up-regulated expression of specific gene on the expression plasmid pEV stimulated by the assistant plasmid pI in the dual-plasmid interactions. Further investigations demonstrated the essential roles of the promoters of the expression plasmid pEV and the CEN/ARS element of the assistant plasmid pI in the dual-plasmid interactions. Combined with the results of predicted nucleosome occupancy, a response model of interaction based on the key T(n)C and CEN/ARS element was established. Moreover, the transformation order of the two plasmids significantly affected the response effect, implying the dominance of plasmid pI in the dual-plasmid interactions. Our finding first demonstrated that dual plasmids regulate the gene expression through spatial interactions at DNA sequences level, which provides a new perspective for the development of microbial cell factories in future.

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.