What is the role of microbial biotechnology and genetic engineering in medicine?

Microbial products are essential for developing various therapeutic agents, including antibiotics, anticancer drugs, vaccines, and therapeutic enzymes. Genetic engineering techniques, functional genomics, and synthetic biology unlock previously uncharacterized natural products. This review highlights major advances in microbial biotechnology, focusing on gene-based technologies for medical applications.

Thermophilic ß-mannanases from bacteria: production, resources, structural features and bioengineering strategies.

ß-mannanases are pivotal enzymes that cleave the mannan backbone to release short chain mannooligosaccharides, which have tremendous biotechnological applications including food/feed, prebiotics and biofuel production. Due to the high temperature conditions in many industrial applications, thermophilic mannanases seem to have great potential to overcome the thermal impediments. Thus, structural analysis of thermostable ß-mannanases is extremely important, as it could open up new avenues for genetic engineering, and protein engineering of these enzymes with enhanced properties and catalytic efficiencies. Under this scope, the present review provides a state-of-the-art discussion on the thermophilic ß-mannanases from bacterial origin, their production, engineering and structural characterization. It covers broad insights into various molecular biology techniques such as gene mutagenesis, heterologous gene expression, and protein engineering, that are employed to improve the catalytic efficiency and thermostability of bacterial mannanases for potential industrial applications. Further, the bottlenecks associated with mannanase production and process optimization are also discussed. Finally, future research related to bioengineering of mannanases with novel protein expression systems for commercial applications are also elaborated.

Essential factors, advanced strategies, challenges, and approaches involved for efficient expression of recombinant proteins in Escherichia coli.

Producing recombinant proteins is a major accomplishment of biotechnology in the past century. Heterologous hosts, either eukaryotic or prokaryotic, are used for the production of these proteins. The utilization of microbial host systems continues to dominate as the most efficient and affordable method for biotherapeutics and food industry productions. Hence, it is crucial to analyze the limitations and advantages of microbial hosts to enhance the efficient production of recombinant proteins on a large scale. E. coli is widely used as a host for the production of recombinant proteins. Researchers have identified certain obstacles with this host, and given the growing demand for recombinant protein production, there is an immediate requirement to enhance this host. The following review discusses the elements contributing to the manifestation of recombinant protein. Subsequently, it sheds light on innovative approaches aimed at improving the expression of recombinant protein. Lastly, it delves into the obstacles and optimization methods associated with translation, mentioning both cis-optimization and trans-optimization, producing soluble recombinant protein, and engineering the metal ion transportation. In this context, a comprehensive description of the distinct features will be provided, and this knowledge could potentially enhance the expression of recombinant proteins in E. coli.

Advances in recombinant protease production: current state and perspectives.

Proteases, enzymes that catalyze the hydrolysis of peptide bonds in proteins, are important in the food industry, biotechnology, and medical fields. With increasing demand for proteases, there is a growing emphasis on enhancing their expression and production through microbial systems. However, proteases' native hosts often fall short in high-level expression and compatibility with downstream applications. As a result, the recombinant production of proteases has become a significant focus, offering a solution to these challenges. This review presents an overview of the current state of protease production in prokaryotic and eukaryotic expression systems, highlighting key findings and trends. In prokaryotic systems, the Bacillus spp. is the predominant host for proteinase expression. Yeasts are commonly used in eukaryotic systems. Recent advancements in protease engineering over the past five years, including rational design and directed evolution, are also highlighted. By exploring the progress in both expression systems and engineering techniques, this review provides a detailed understanding of the current landscape of recombinant protease research and its prospects for future advancements.

Protocols for Building and Producing High Diversity Peptide Phage Display Libraries.

Phage display is an important technology to study protein-protein interaction and protein evolution, with applications in basic science and applied biotechnology, such as drug discovery and the development of targeted therapies. However, in order to be successful during a phage display screening, it is paramount to have good phage libraries. Here, we described detailed procedures to generate peptide phage display libraries with high diversity and billions of transformants.

Life cycle assessment and techno-economic analysis of wood-based biorefineries for cellulosic ethanol production.

Poplar is widely used in the paper industry and accompanied by abundant branches waste, which is potential feedstock for bioethanol production. Acid-chlorite pretreatment can selectively remove lignin, thereby significantly increasing enzymatic efficiency. Moreover, lignin residues valorization via gasification-syngas fermentation can achieve higher fuel yield. Herein, environmental and economic aspects were conducted to assess technological routes, which guides further process optimization. Life cycle assessment results show that wood-based biorefineries especially coupling scenarios have significant advantages in reducing global warming potential in contrast to fossil-based automotive fuels. Normalization results indicate that acidification potential surpasses other indicators as the primary impact category. In terms of economic feasibility, coupling scenarios present better investment prospects. Bioethanol yield is the most critical factor affecting market competitiveness. Minimum ethanol selling price below ethanol international market price is promising with higher-levels technology. Further work should be focused on technological breakthrough, consumable reduction or replacement.

Biocompounds from wastewater-grown microalgae: a review of emerging cultivation and harvesting technologies.

Microalgae biomass has attracted attention as a feedstock to produce biofuels, biofertilizers, and pigments. However, the high production cost associated with cultivation and separation stages is a challenge for the microalgae biotechnology application on a large scale. A promising approach to overcome the technical-economic limitations of microalgae production is using wastewater as a nutrient and water source for cultivation. This strategy reduces cultivation costs and contributes to valorizing sanitation resources. Therefore, this article presents a comprehensive literature review on the status of microalgae biomass cultivation in wastewater, focusing on production strategies and the accumulation of valuable compounds such as lipids, carbohydrates, proteins, fatty acids, and pigments. This review also covers emerging techniques for harvesting microalgae biomass cultivated in wastewater, discussing the advantages and limitations of the process, as well as pointing out the main research opportunities. The novelty of the study lies in providing a detailed analysis of state-of-the-art and potential advances in the cultivation and harvesting of microalgae, with a special focus on the use of wastewater and implementing innovative strategies to enhance productivity and the accumulation of compounds. In this context, the work aims to guide future research concerning emerging technologies in the field, emphasizing the importance of innovative approaches in cultivating and harvesting microalgae for advancing knowledge and practical applications in this area.

Metagenomic analysis revealed the potential of lactic acid bacteria in improving natural saline-alkali land / El análisis metagenómico reveló el potencial de las bacterias del ácido láctico para mejorar las tierras salino-álcalis naturales

Management and improving saline-alkali land is necessary for sustainable agricultural development. We conducted a field experiment to investigate the effects of spraying lactic acid bacteria (LAB) on the cucumber and tomato plant soils. Three treatments were designed, including spraying of water, viable or sterilized LAB preparations to the soils of cucumber and tomato plants every 20 days. Spraying sterilized or viable LAB could reduce the soil pH, with a more obvious effect by using viable LAB, particularly after multiple applications. Metagenomic sequencing revealed that the soil microbiota in LAB-treated groups had higher alpha-diversity and more nitrogen-fixing bacteria compared with the water-treated groups. Both viable and sterilized LAB, but not water application, increased the complexity of the soil microbiota interactive network. The LAB-treated subgroups were enriched in some KEGG pathways compared with water or sterilized LAB subgroups, such as environmental information processing–related pathways in cucumber plant; and metabolism-related pathways in tomato plant, respectively. Redundancy analysis revealed association between some soil physico-chemical parameters (namely soil pH and total nitrogen) and bacterial biomarkers (namely Rhodocyclaceae, Pseudomonadaceae, Gemmatimonadaceae, and Nitrosomonadales). Our study demonstrated that LAB is a suitable strategy for decreasing soil pH and improving the microbial communities in saline-alkali land.(AU)

Application of extremophile cell factories in industrial biotechnology.

Due to the extreme living conditions, extremophiles have unique characteristics in morphology, structure, physiology, biochemistry, molecular evolution mechanism and so on. Extremophiles have superior growth and synthesis capabilities under harsh conditions compared to conventional microorganisms, allowing for unsterilized fermentation processes and thus better performance in low-cost production. In recent years, due to the development and optimization of molecular biology, synthetic biology and fermentation technology, the identification and screening technology of extremophiles has been greatly improved. In this review, we summarize techniques for the identification and screening of extremophiles and review their applications in industrial biotechnology in recent years. In addition, the facts and perspectives gathered in this review suggest that next-generation industrial biotechnology (NGIBs) based on engineered extremophiles holds the promise of simplifying biofuturing processes, establishing open, non-sterilized continuous fermentation production systems, and utilizing low-cost substrates to make NGIBs attractive and cost-effective bioprocessing technologies for sustainable manufacturing.

Breeding of ornamental orchids with focus on Phalaenopsis: current approaches, tools, and challenges for this century.

Ornamental orchid breeding programs have been conducted to develop commercially valuable cultivars with improved characteristics of commercial interest, such as size, flower color, pattern, shape, and resistance to pathogens. Conventional breeding, including sexual hybridization followed by selection of desirable characteristics in plants, has so far been the main method for ornamental breeding, but other techniques, including mutation induction by polyploidization and gamma irradiation, and biotechnological techniques, such as genetic transformation, have also been studied and used in ornamental breeding programs. Orchids are one of the most commercially important families in floriculture industry, having very particular reproductive biology characteristics and being a well-studied group of ornamentals in terms of genetic improvement. The present review focuses on the conventional and biotechnological techniques and approaches specially employed in breeding Phalaenopsis orchids, the genus with highest worldwide importance as an ornamental orchid, highlighting the main limitations and strengths of the approaches. Furthermore, new opportunities and future prospects for ornamental breeding in the CRISPR/Cas9 genome editing era are also discussed. We conclude that conventional hybridization remains the most used method to obtain new cultivars in orchids. However, the emergence of the first biotechnology-derived cultivars, as well as the new biotechnological tools available, such as CRISPR-Cas9, rekindled the full potential of biotechnology approaches and their importance for improve ornamental orchid breeding programs.

Unlocking the potential of cotton stalk as a renewable source of cellulose: A review on advancements and emerging applications.

Cotton stalk (CS) is a global agricultural residue, with an annual production of approximately 50 million tons, albeit with limited economic significance. The utilization of cellulose derived from CS has gained significant attention in green nanomaterial technologies. This interest stems from its unique properties, including biocompatibility, low density, minimal thermal expansion, eco-friendliness, renewability, and its potential as an alternative source for chemicals, petroleum, and biofuels. In this review, we delve into various extraction and characterization methods, the physicochemical attributes, recent advancements, and the applications of cellulose extracted from CS. Notably, the steam explosion method has proven to yield the highest cellulose content (82 %) from CS. Moreover, diverse physicochemical properties of cellulose can be obtained through different extraction techniques. Sulfuric acid hydrolysis, for instance, yields nanocrystalline cellulose fibers measuring 10-100 nm in width and 100-850 nm in length. Conversely, the steam explosion method yields cellulose fibers with dimensions of 10.7 µm in width and 1.2 mm in length. CS-derived products, including biochar, aerogel, dye adsorbents, and reinforcement fillers, find applications in various industries, such as environmental remediation and biodegradable packaging. This is primarily due to their ready availability, cost-effectiveness, and sustainable nature.

Biphasic pretreatment excels over conventional sulfuric acid in pinewood biorefinery: An environmental analysis.

This study assesses the environmental impact of pine chip-based biorefinery processes, focusing on bioethanol, xylonic acid, and lignin production. A cradle-to-gate Life Cycle Assessment (LCA) is employed, comparing a novel biphasic pretreatment method (p-toluenesulfonic acid (TsOH)/pentanol, Sc-1) with conventional sulfuric acid pretreatment (H2SO4, Sc-2). The analysis spans biomass handling, pretreatment, enzymatic hydrolysis, yeast fermentation, and distillation. Sc-1 yielded an environmental impact of 1.45E+01 kPt, predominantly affecting human health (96.55%), followed by ecosystems (3.07%) and resources (0.38%). Bioethanol, xylonic acid, and lignin contributed 32.61%, 29.28%, and 38.11% to the total environmental burdens, respectively. Sc-2 resulted in an environmental burden of 1.64E+01 kPt, with a primary impact on human health (96.56%) and smaller roles for ecosystems (3.07%) and resources (0.38%). Bioethanol, xylonic acid, and lignin contributed differently at 22.59%, 12.5%, and 64.91%, respectively. Electricity generation was predominant in both scenarios, accounting for 99.05% of the environmental impact, primarily driven by its extensive usage in biomass handling and pretreatment processes. Sc-1 demonstrated a 13.05% lower environmental impact than Sc-2 due to decreased electricity consumption and increased bioethanol and xylonic acid outputs. This study highlights the pivotal role of pretreatment methods in wood-based biorefineries and underscores the urgency of sustainable alternatives like TsOH/pentanol. Additionally, adopting greener electricity generation, advanced technologies, and process optimization are crucial for reducing the environmental footprint of waste-based biorefineries while preserving valuable bioproduct production.

Intellectual Property Rights: Protection of Biotechnological Inventions in India.

The current Intellectual Property Rights (IPR) framework supports the commercialization of seed improvement, monoculture, and the patent protection of novel plant varieties, microorganisms, and genetically modified animals. As a consequence, our rich biogenetic diversity is irreversibly dissipating. However, we need to figure out how to create a methodology for elective choices that will achieve harmony between the official Intellectual Property (IP) structure and maintainable biodiversity components. The majority of the biotechnology sector's programmes in India are managed by the Department of Biotechnology. It is under the Ministry of Science and Technology. Its goals are to provide services in the fields of study, infrastructure, human resource development, biotechnology popularisation, industry promotion, and establishment of centres of excellence. Implementation of practise biosafety regulations for genetically modified organisms, recombinant DNA products, and programmes is based on biotechnology for the good of society. This creates an information network for India's bioinformatics mission in the local, national, and worldwide scientific community.

Application of response surface Methodology coupled with Artificial Neural network and genetic algorithm to model and optimize symbiotic interactions between Chlorella vulgaris and Stutzerimonas stutzeri strain J3BG for chlorophyll accumulation.

Research on microalgae has surged due to its diverse biotechnological applications and capacity for accumulating bioactive compounds. Despite considerable advancements, microalgal cultivation remains costly, prompting efforts to reduce expenses while enhancing productivity. This study proposes a cost-effective approach through the coculture of microalgae and bacteria, exploiting mutualistic interactions. An engineered consortium of Chlorella vulgaris and Stutzerimonas stutzeri strain J3BG demonstrated biofilm-like arrangements, indicative of direct cell-to-cell interactions and metabolite exchange. Strain J3BG's enzymatic characterization revealed amylase, lipase, and protease production, sustaining mutual growth. Employing Response Surface Methodology (RSM), Artificial Neural Network (ANN), and Genetic Algorithm (GA) in a hybrid modeling approach resulted in a 2.1-fold increase in chlorophyll production. Optimized conditions included a NaNO3 concentration of 128.52 mg/l, a 1:2 (Algae:Bacteria) ratio, a 6-day cultivation period, and a pH of 5.4, yielding 10.92 ± 0.88 mg/l chlorophyll concentration.

Self-assembling peptides: Perspectives regarding biotechnological applications and vaccine development.

Self-assembly involves a set of molecules spontaneously interacting in a highly coordinated and dynamic manner to form a specific supramolecular structure having new and clearly defined properties. Many examples of this occur in nature and many more came from research laboratories, with their number increasing every day via ongoing research concerning complex biomolecules and the possibility of harnessing it when developing new applications. As a phenomenon, self-assembly has been described on very different types of molecules (biomolecules including), so this review focuses on what is known about peptide self-assembly, its origins, the forces behind it, how the properties of the resulting material can be tuned in relation to experimental considerations, some biotechnological applications (in which the main protagonists are peptide sequences capable of self-assembly) and what is yet to be tuned regarding their research and development.

Tools for investigating O-GlcNAc in signaling and other fundamental biological pathways.

Cells continuously fine-tune signaling pathway proteins to match nutrient and stress levels in their local environment by modifying intracellular proteins with O-linked N-acetylglucosamine (O-GlcNAc) sugars, an essential process for cell survival and growth. The small size of these monosaccharide modifications poses a challenge for functional determination, but the chemistry and biology communities have together created a collection of precision tools to study these dynamic sugars. This review presents the major themes by which O-GlcNAc influences signaling pathway proteins, including G-protein coupled receptors, growth factor signaling, mitogen-activated protein kinase (MAPK) pathways, lipid sensing, and cytokine signaling pathways. Along the way, we describe in detail key chemical biology tools that have been developed and applied to determine specific O-GlcNAc roles in these pathways. These tools include metabolic labeling, O-GlcNAc-enhancing RNA aptamers, fluorescent biosensors, proximity labeling tools, nanobody targeting tools, O-GlcNAc cycling inhibitors, light-activated systems, chemoenzymatic labeling, and nutrient reporter assays. An emergent feature of this signaling pathway meta-analysis is the intricate interplay between O-GlcNAc modifications across different signaling systems, underscoring the importance of O-GlcNAc in regulating cellular processes. We highlight the significance of O-GlcNAc in signaling and the role of chemical and biochemical tools in unraveling distinct glycobiological regulatory mechanisms. Collectively, our field has determined effective strategies to probe O-GlcNAc roles in biology. At the same time, this survey of what we do not yet know presents a clear roadmap for the field to use these powerful chemical tools to explore cross-pathway O-GlcNAc interactions in signaling and other major biological pathways.

Transformations of bamboo into bioethanol through biorefinery.

The increasing demand for energy has prompted scholars to research alternative energy sources. Bamboo is a species of woody perennial grass that belongs to the Gramineae family and the Bambuseae subfamily. It could be considered a possible lignocellulosic substrate for the production of bioethanol due to its favourable environmental effects and increased yearly biomass yield. Non-renewable fossil fuels cannot provide enough energy to meet the needs of contemporary societies. Among the various alternative energy sources, bioethanol has drawn a lot of attention from people all around the world. This paper reviews the cost and process parameters for the synthesis of bioethanol from bamboo. This review aims to increase the effectiveness of the entire ethanol production process by focusing on pretreatment, enzymatic hydrolysis, and fermentation. The emphasis of this review is on the efficient process for producing bioethanol while maintaining environmental sustainability. When compared to other NaOH pretreatment techniques, bamboo substrates prepared with NaOH and ultra-high-pressure explosion (UHPE) exhibit higher enzymatic hydrolyzability when processed under optimal conditions, such as 100 MPa, 121 °C, and 70 rpm for 2 h, yielding 89.7-95.1% ethanol after 24 h. The article lists the bamboo species responsible for creating each product, making it straightforward for producers to study and select the species based on whatever value-added product they wish to produce bioethanol with different parameters.

Construction of a Stable Expression System Based on the Endogenous hbpB/hbpC Toxin-Antitoxin System of Halomonas bluephagenesis.

Halomonas bluephagenesis is a halophilic bacterium capable of efficiently producing polyhydroxyalkanoates and other valuable chemicals through high salinity open fermentation, offering an appealing platform for next-generation industrial biotechnology. Various techniques have been developed to engineer Halomonas bluephagenesis, each with its inherent shortcomings. Genome editing methods often entail complex and time-consuming processes, while flexible expression systems relying on plasmids necessitate the use of antibiotics. In this study, we developed a stable recombinant plasmid vector, pHbPBC, based on a novel hbpB/hbpC toxin-antitoxin system found within the endogenous plasmid of Halomonas bluephagenesis. Remarkably, pHbPBC exhibited exceptional stability during 7 days of continuous subculture, eliminating the need for antibiotics or other selection pressures. This stability even rivaled genomic integration, all while achieving higher levels of heterologous expression. Our research introduces a novel approach for genetically modifying and harnessing nonmodel halophilic bacteria, contributing to the advancement of next-generation industrial biotechnology.