Field Of Biotechnology Research Articles

A perspective of lipid nanoparticles for RNA delivery

AbstractOver the last two decades, lipid nanoparticles (LNPs) have evolved as an effective biocompatible and biodegradable RNA delivery platform in the fields of nanomedicine, biotechnology, and drug delivery. They are novel bionanomaterials that can be used to encapsulate a wide range of biomolecules, such as mRNA, as demonstrated by the current successes of COVID‐19 mRNA vaccines. Therefore, it is important to provide a perspective on LNPs for RNA delivery, which further offers useful guidance for researchers who want to work in the RNA‐based LNP field. This perspective first summarizes the approaches for the preparation of LNPs, followed by the introduction of the key characterization parameters. Then, the in vitro cell experiments to study LNP performance, including cell selection, cell viability, cellular association/uptake, endosomal escape, and their efficacy, were summarized. Finally, the in vivo animal experiments in the aspects of animal selection, administration, dosing and safety, and their therapeutic efficacy were discussed. The authors hope this perspective can offer valuable guidance to researchers who enter the field of RNA‐based LNPs and help them understand the crucial parameters that RNA‐based LNPs demand.

Development of an Advanced Biology Learning Website in the Fields of Biotechnology, Biochemistry, and Biomedicine with the STEAM Approach

The growing industry has given birth to various innovations, one of which is the emergence of three new branches of knowledge in biology: biotechnology, biochemistry, and biomedicine. However, this branch of science has not been explored in biology education for students. Technology, in the form of online learning platforms, can address current challenges. STEAM-based biology learning can increase student engagement, motivate creative exploration, and stimulate critical thinking. This study aims to develop an advanced biology learning platform and test the feasibility of the learning media. This study follows the analysis, design, development, implementation, and evaluation (ADDIE) model. The results of this study have led to the development of the “Advanced Biology” website, which contains various exciting features for students. This feature includes a homepage display, learning content display, case-based learning, and research project display. This platform provides engaging educational videos and comprehensive learning materials. The case studies on this site offer an overview of problem-solving and the current state of the industry. This media validates learning materials, media, and technology in the ideal category.

Non-toxic orange carbon dots stimulate photosynthesis and CO2 assimilation in hydroponically cultivated green beans (Phaseolus vulgaris).

Continuous increasing leaf photosynthesis may enhance plant yield. As an evolutionary property, plants use less photosynthetic capacity than is theoretically possible. Plant nanobionics is a bioengineering field that improves plant functions using nanoparticles. We applied orange carbon dots (o-CDs) onto the foliage of green beans (Phaseolus vulgaris ) grown in hydroponics to improve their photosynthetic performance and CO2 assimilation. Photosynthesis parameters, photosynthetic pigments content, total phenolic content (TPC) and antioxidative activity (TAA) were measured. Results show that photosynthetic pigments remained unchanged, while photosynthesis was improved. Both o-CDs concentrations decreased TPC and TAA. The light response curve showed higher CO2 assimilation at both o-CDs concentrations, particularly at lower light intensity. Correlation analysis confirmed increased CO2 binding and assimilation at 1mg L-1 . This study demonstrated the potential of using o-CDs as a safe biostimulator through photosynthesis increase and CO2 assimilation without toxic effects on plants. This may stimulate yield increase that paves the way for their agricultural application.

Diversity analysis of sea anemone peptide toxins in different tissues of Heteractis crispa based on transcriptomics

Peptide toxins found in sea anemones venom have diverse properties that make them important research subjects in the fields of pharmacology, neuroscience and biotechnology. This study used high-throughput sequencing technology to systematically analyze the venom components of the tentacles, column, and mesenterial filaments of sea anemone Heteractis crispa, revealing the diversity and complexity of sea anemone toxins in different tissues. A total of 1049 transcripts were identified and categorized into 60 families, of which 91.0% were proteins and 9.0% were peptides. Of those 1049 transcripts, 416, 291, and 307 putative proteins and peptide precursors were identified from tentacles, column, and mesenterial filaments respectively, while 428 were identified when the datasets were combined. Of these putative toxin sequences, 42 were detected in all three tissues, including 33 proteins and 9 peptides, with the majority of peptides being ShKT domain, β-defensin, and Kunitz-type. In addition, this study applied bioinformatics approaches to predict the family classification, 3D structures, and functional annotation of these representative peptides, as well as the evolutionary relationships between peptides, laying the foundation for the next step of peptide pharmacological activity research.

Evaluation of Heavy Metal Concentrations in Enzyme Embedded Poly Lactic Acid (EPLA) by ICP-OES: Lipase from Candida Rugosa.

Abstract: Enzymes like lipases are essential for catalyzing important reactions in the fields of biotechnology and industrial operations. For quality control and safety evaluations, however, the quantification of these contaminants is crucial because the presence of heavy metals in enzyme preparations can negatively affect their activity and stability. Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES), a key component of this investigation, is used to measure the levels of heavy metals in lipase enzyme preparations embedded in polylactic acid (PLA). The lipase enzyme being studied comes from Candida rugosa, which is well-known for its considerable industrial applications. According to preliminary findings, the lipase enzyme is successfully protected from external heavy metal pollutants by the encapsulating technique, maintaining its enzymatic activity and stability. The lipase-PLA composite had only minimal quantities of heavy metals, according to the ICP-OES study. The research's conclusions show great potential for the creation of reliable and contaminant-free lipase enzyme preparations, which will increase their suitability for usage in a variety of industrial processes and encourage the use of biodegradable polymers, supporting environmental sustainability. By highlighting the significance of quality control and safety assessment through the determination of heavy metal concentrations, this research contributes to the larger field of enzyme biotechnology. It also highlights the potential of Candida Rugosa lipase implanted in PLA matrices for eco-friendly and sustainable biocatalytic applications in sectors ranging from food and pharmaceuticals to biofuel generation and other biodegradable polymers.

Nanotechnology and The Use of Nanoparticles and Its Effect on Wheat Growing

The rapid increase in the human population and the improvement in the living standards of many countries in the world cause the increasing supply of agricultural land to be unmet. Therefore, soil fertility is gaining importance day by day and soil texture is becoming a strategic element. With the development of nanotechnology, the usability of nanoparticles in many fields, especially in agriculture, has started to be investigated or discussed. Studies in agricultural nanotechnology have generally focused on using less pesticides, increasing yields or developing stress-resistant crops. Some studies in this field have started to yield positive results. However, more studies are needed for nanotechnology to be used in agriculture. Because deciding on the use of nanoparticles is an issue that can be reached in the long term. In recent years, the application of nanotechnology-based applications in agriculture is one of the strategies that attract the attention of researchers. Nanotechnology explores a wide spectrum of applications in the fields of biotechnology and agriculture sector, creating a broad spectrum for various applications. Nanotechnology has become a new technology that can be used in various industries such as industry, medicine, food science and safety, smart packaging and agriculture. Nanotechnology, which has been used in many stages of agriculture in recent years, has been widely applied in product production, development, processing, packaging, storage and transportation, and has brought about major changes in food and agricultural systems. It is also used to increase plant resistance and to control the efficacy and safety of pesticides and fertilizers. Thus, the application of nanostructured materials designed for sustainable crop production reduces nutrient losses, suppresses diseases and increases yields. In this study, the strengths, weaknesses, opportunities and targets for the use of nanotechnology and nanoparticles in wheat agriculture were identified and presented.

Molecular imprinted polymers: important advances in biochemistry, biomedical and biotechnology

AbstractMolecularly imprinted polymers (MIPs) are handmade receptors that mimic the binding of natural antibodies. In other words, MIPs can selectively bind to the target molecule and qualify as bio-inspired synthetic materials. Today, MIPs are used extensively and are being developed further for biological applications. High cost and time consuming techniques are compelling factors for the field of biochemistry, biomedicine and biotechnology (3B), and there is an urgent need for an alternative, cheap, easy to produce, fast and effective method in these fields. MIPs stand out as a promising way for this purpose. MIPs have superiorities such as specific recognition specificity, excellent sensitivity selectivity and reusability. From this point of view, we examined MIP-related drug delivery studies, cell recognition, enzyme applications, in vivo applications, and applications for some important biomolecules. The aim of this review is to compile the utilizations, advantages, important developments and future expectations of MIPs for the fields of 3B.

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.

Novel CRISPR/SpRY system for rapid detection of CRISPR/Cas-mediated gene editing in rice

The gene editing technology represented by clustered rule-interspersed short palindromic repeats (CRISPR)/Cas9 has developed as a common tool in the field of biotechnology. Many gene-edited products in plant varieties have recently been commercialized. However, the rapid on-site visual detection of gene-edited products without instrumentation remains challenging. This study aimed to develop a novel and efficient method, termed the CRISPR/SpRY detection platform, for the rapid screening of CRISPR/Cas9-induced mutants based on CRISPR/SpRY-mediated in vitro cleavage using rice (Oryza sativa L.) samples genetically edited at the TGW locus as an example. We designed the workflow of the CRISPR/SpRY detection platform and conducted a feasibility assessment. Subsequently, we optimized the reaction system of CRISPR/SpRY, and developed a one-pot CRISPR/SpRY assay by integrating recombinase polymerase amplification (RPA). The sensitivity of the method was further verified using recombinant plasmids. The proposed method successfully identified various types of mutations, including insertions, deletions (indels), and nucleotide substitutions, with excellent sensitivity. Finally, the applicability of this method was validated using different rice samples. The entire process was completed in less than an hour, with a limit of detection as low as 1%. Compared with previous methods, our approach is simple to operate, instrumentation-free, cost-effective, and time-efficient. The primary significance lies in the liberation of our developed system from the limitations imposed using protospacer adjacent motif sequences. This expands the scope and versatility of the CRISPR-based detection platform, making it a promising and groundbreaking platform for detecting mutations induced by gene editing.

Purification and Biochemical Characterization of Phytase Enzyme from Lactobacillus brevis Isolated from Fresh Kashar Cheese

In the last 20 years, phytase enzyme has attracted the attention of scientists in the fields of environmental protection, nutrition and biotechnology. Myo-inositol hexaphosphate phosphohydrolase (phytase), which is a type of phosphatase enzyme, catalyzes the hydrolysis of phytate into less phosphorylated inorganic phosphates and phytate. Phytases of microbial origin are widely used in biotechnological applications (paper industry, feed industry, food industry and soil improvement).
 In the present study, phytase enzyme was partially purified from Lactobacillus brevis NM-34 strain isolated from fresh kashar cheese and the pH and temperature values at which the enzyme showed optimum activity were determined. L. brevis NM-34 showed a phytase activity of 243.80 U/mL as a result of ammonium sulfate precipitation. In the ammonium sulfate range (40-60%), where the highest phytase activity was observed, the protein concentration was measured as 0.989 mg/mL. Km and Vmax values of phytase enzyme were determined as 0.0146 mM and 1.6 µmol/min, respectively. The pH and temperature values at which the partially purified phytase showed optimum activity were found to be pH 5 and 50 °C, respectively. Based on the findings from our research, the enzyme purified from this bacterium was found to have unique properties that make it suitable for use in industrial applications.

Multi-Functional Integration of Phosphor, Initiator, and Crosslinker for the Photo-Polymerization of Flexible Phosphorescent Polymer Gels.

A general approach to constructing room temperature phosphorescence (RTP) materials involves the incorporation of a phosphorescent emitter into a rigid host or polymers with high glass transition temperature. However, these materials often suffer from poor processability and suboptimal mechanical properties, limiting their practical applications. In this work, we developed benzothiadiazole-based dialkene (BTD-HEA), a multifunctional phosphorescent emitter with a remarkable yield of intersystem crossing (ΦISC, 99.83 %). Its high triplet exciton generation ability and dialkene structure enable BTD-HEA to act as a photoinitiator and crosslinker, efficiently initiating the polymerization of various monomers within 120 seconds. A range of flexible phosphorescence gels, including hydrogels, organogels, ionogels, and aerogels were fabricated, which exhibit outstanding stretchability and recoverability. Furthermore, the unique fluorescent-phosphorescent colorimetric properties of the gels provide a more sensitive method for the visual determination of the polymerization process. Notably, the phosphorescent emission intensity of the hydrogel can be increased by the formation of ice, allowing for the precise detection of hydrogel freezing. The versatility of this emitter paves the way for fabricating various flexible phosphorescence gels with diverse morphologies using microfluidics, film-shearing, roll coating process, and two/three-dimensional printing, showcasing its potential applications in the fields of bioimaging and bioengineering.

Multi‐Functional Integration of Phosphor, Initiator, and Crosslinker for the Photo‐Polymerization of Flexible Phosphorescent Polymer Gels

AbstractA general approach to constructing room temperature phosphorescence (RTP) materials involves the incorporation of a phosphorescent emitter into a rigid host or polymers with high glass transition temperature. However, these materials often suffer from poor processability and suboptimal mechanical properties, limiting their practical applications. In this work, we developed benzothiadiazole‐based dialkene (BTD‐HEA), a multifunctional phosphorescent emitter with a remarkable yield of intersystem crossing (ΦISC, 99.83 %). Its high triplet exciton generation ability and dialkene structure enable BTD‐HEA to act as a photoinitiator and crosslinker, efficiently initiating the polymerization of various monomers within 120 seconds. A range of flexible phosphorescence gels, including hydrogels, organogels, ionogels, and aerogels were fabricated, which exhibit outstanding stretchability and recoverability. Furthermore, the unique fluorescent‐phosphorescent colorimetric properties of the gels provide a more sensitive method for the visual determination of the polymerization process. Notably, the phosphorescent emission intensity of the hydrogel can be increased by the formation of ice, allowing for the precise detection of hydrogel freezing. The versatility of this emitter paves the way for fabricating various flexible phosphorescence gels with diverse morphologies using microfluidics, film‐shearing, roll coating process, and two/three‐dimensional printing, showcasing its potential applications in the fields of bioimaging and bioengineering.

Preface: 3rd International Conference on Biotechnology, Life Science and Medical Engineering (BLSME 2024)

This conference proceedings volume contains the written versions of the contributions presented during the 2024 3rd International Conference on Biotechnology, Life Science and Medical Engineering (BLSME 2024), it had been held in Jeju, South Korea during January 13-14, 2024. This conference is the premier forum for the presentation of new advances and research results in the fields of biotechnology, bioengineering, biochemistry, preclinical medicine, clinical medicine, pharmacology, life science, medical engineering, and medicine technology.
 The conference provided a forum for discussion and sharing between lecturers, researchers, academics, practitioners, communities, and specialists, from universities, research institutes, corporations, industries, the government, and the public. The conference has been a great opportunity for participants coming from many countries to present and discuss topics in their respective research areas.
 BLSME 2024 conference included 4 plenary sessions and invited speakers, and oral and poster presentations to allow as many as possible attendances to present their scientific results on the different fields. The manuscripts which have presented were published under the peer-reviewed process.
 We would love to express our gratitude for all the participants, keynote speakers, and guests of honor, for their effective contribution in our conference. Our ongoing thankfulness is presented for our guest editors and speakers. Finally, we would love to express our gratefulness for all our sponsors, partners, and colleagues, we hope to collaborate with them in future activities.
 Chairman of Conference Committee
 Jeju, South Korea

Marine thraustochytrid: exploration from taxonomic challenges to biotechnological applications

Thraustochytrids, as a distinct group of heterotrophic protists, have garnered considerable attention owing to their remarkable adaptability in extreme marine environments, pronounced capacity for metabolic regulation and prolific production of high-value metabolites. The taxonomic classification of these microorganisms presents a substantial challenge due to the variability in morphological characteristics under different culture conditions. And this undermines the efficacy of traditional classification systems on physiological and biochemical traits. The establishment of a polyphasic taxonomic system integrating genomic characteristics in the future will provide new avenues for more accurate classification and identification. Thraustochytrids can effectively accumulate bioactive substances such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), squalene and carotenoids. Through fermentation optimization and genetic modification, scientists have significantly enhanced the production of these metabolites. Moreover, the application of thraustochytrids in aquaculture, poultry and livestock feed has significantly improved animal growth and physiological indicators meanwhile increasing their DHA content. Natural bioactive substances in thraustochytrids, such as terpenoid compounds with antioxidant properties, have been proposed for application in the cosmetics industry. In the field of pharmacology, thraustochytrids have shown certain anti-inflammatory and anti-cancer activities and provide potential for the development of new oral vaccines. Additionally, they can degrade various industrial and agricultural wastes for growth and fatty acid production, demonstrating their potential in environmental bioremediation. Therefore, thraustochytrids not only exhibit tremendous application potential in the field of biotechnology, but also hold significant value in environmental protection and commercialization.

A low-cost and eco-friendly recombinant protein expression system using copper-containing industrial wastewater.

The development of innovative methods for highly efficient production of recombinant proteins remains a prominent focus of research in the biotechnology field, primarily due to the fact that current commercial protein expression systems rely on expensive chemical inducers, such as isopropyl β-D-thiogalactoside (IPTG). In our study, we designed a novel approach for protein expression by creating a plasmid that responds to copper. This specialized plasmid was engineered through the fusion of a copper-sensing element with an optimized multiple cloning site (MCS) sequence. This MCS sequence can be easily customized by inserting the coding sequences of target recombinant proteins. Once the plasmid was generated, it was introduced into an engineered Escherichia coli strain lacking copA and cueO. With this modified E. coli strain, we demonstrated that the presence of copper ions can efficiently trigger the induction of recombinant protein expression, resulting in the production of active proteins. Most importantly, this expression system can directly utilize copper-containing industrial wastewater as an inducer for protein expression while simultaneously removing copper from the wastewater. Thus, this study provides a low-cost and eco-friendly strategy for the large-scale recombinant protein production. To the best of our knowledge, this is the first report on the induction of recombinant proteins using industrial wastewater.

Non-similar analysis of bio-convective micropolar nanofluid flow including gyrotactic microorganisms across a stretched geometry

This study explores the novel interaction of gyrotactic microorganisms within a two-dimensional bioconvective micropolar nanofluid flow along a stretched surface, employing the Tiwari-Das nanofluid model. Uniquely, blood is modeled as a micropolar fluid incorporating titanium dioxide ( Ti O 2 ) and silver   ( Ag ) nanoparticles. The analysis incorporates influential factors such as viscous dissipation, magnetic field, and heat source effects. By converting the control system into dimensionless nonlinear coupled differential equations, a complete understanding of the system is attained through the application of the local non-similarity approach and bvp4c (MATLAB tool). Particularly, the study provides valuable insight into the influence of dimensionless control parameters on key profiles such as velocity, microrotation, temperature, concentration, and microbial profiles. The findings show that temperature, microrotation, and velocity profiles are all improved by raising the values of micropolar parameters. Furthermore, the microbe profile shows an inverse connection with the Lewis number and a positive association with the Peclet number, whereas the concentration profile declines with increasing Schmidt number values. Furthermore, the study provides a quantitative analysis through tables, revealing variations in the drag coefficient and Nusselt number. This innovative exploration sheds light on the unique behavior of micropolar bioconvective nanofluids, offering valuable contributions to the field of fluid dynamics and bioengineering.

Exploring Bacillus subtilis: Ecology, biotechnological applications, and future prospects.

From its early identification by Christian Gottfried Ehrenberg to its current prominence in scientific research, Bacillus subtilis (B. subtilis) has emerged as a foundational model organism in microbiology. This comprehensive review delves deep into its genetic, physiological, and biochemical intricacies, revealing a sophisticated cellular blueprint. With the incorporation of advanced techniques such as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 and integrative computational methodologies, the potential applications of B. subtilis span diverse sectors. These encompass its significant contributions to biotechnology, agriculture, and medical fields and its potential for aiding environmental cleanup efforts. Yet, as we move forward, we must grapple with concerns related to safety, ethics, and the practical implementation of our lab findings in everyday scenarios. As our understanding of B. subtilis deepens, it is evident that its contributions will be central to pioneering sustainable solutions for global challenges in the years to come.

Semen sexing and its impact on fertility and genetic gain in cattle.

Semen sexing is among one of the most remarkable inventions of the past few decades in the field of reproductive biotechnology. The urge to produce offspring of a desired sex has remained since traditional times. Researchers have tried many methods for accurate semen sexing, but only the flow cytometry method has proved to be effective for commercial utilization. However, there were always concerns about the effects of sexed semen, especially on fertility and the rate of genetic gain. Some concerns were genuine because of factors such as low semen dosage in sexed semen straws and damage to sperm during the sorting process. Various researchers have conducted numerous studies to find out the effect of sexed semen on fertility and, in this article, we reflect on their findings. Initially, there were comparatively much lower conception rates (∼70% of conventional semen) but, with refinement in technology, this gap is bridging and the use of sexed semen will increase over time. Concerning genetic gain with use of sexed semen, a positive effect on rate of genetic progress with the use of sexed semen has been observed based on various simulation studies, although there has been a mild increase in inbreeding.

Drug delivery using reduction-responsive hydrogel based on carboxyethyl-succinoglycan with highly improved rheological, antibacterial, and antioxidant properties

The development of exopolysaccharide-based polymers is gaining increasing attention in various industrial biotechnology fields for materials such as thickeners, texture modifiers, anti-freeze agents, antioxidants, and antibacterial agents. High-viscosity carboxyethyl-succinoglycan (CE-SG) was directly synthesized from succinoglycan (SG) isolated from Sinorhizobium meliloti Rm 1021, and its structural, rheological, and physiological properties were investigated. The viscosity of CE-SG gradually increased in proportion to the degree of carboxyethylation substitution. In particular, when the molar ratio of SG and 3-chloropropionic acid was 1:100, the viscosity was significantly improved by 21.18 times at a shear rate of 10 s−1. Increased carboxyethylation of SG also improved the thermal stability of CE-SG. Furthermore, the CE-SG solution showed 90.18 and 91.78 % antibacterial effects against Escherichia coli and Staphylococcus aureus and effective antioxidant activity against DPPH and hydroxyl radicals. In particular, CE-SG hydrogels coordinated with Fe3+ ions, which improved both viscosity and rheological properties, while also exhibiting reduction-responsive drug release through 1,4-dithiothreitol. The results of this study suggest that SG derivatives, such as CE-SG, can be used as functional biomaterials in various fields such as food, cosmetics, and pharmaceutical industries.

Bioremediation Studies on Pesticides in Soil by Novel Microorganisms Isolated from Agricultural Fields

Pesticides are organic compounds widely used in agriculture for increasing agricultural yield, soil productivity and to control the insect vectors for prevention of various plants, human and animal epidemics. Pesticides are widely distributed in our ecosystem. Organophosphates are found to be one of the most common group of pesticides found across the world. However, excessive and indiscriminate use of pesticides have generated a number of environmental problems such as release of toxic waste in the environment resulting to contamination of air, water and terrestrial ecosystem. In order to control the harmful effects of pesticides, multiple attempts have been made like the use of physical, chemical, biological and enzymatic approaches to reduce pesticides. However, physical and chemical methods have proven to be inefficient. One of the most promising field of biotechnology that offers a novel approach to degrade the pesticides is bioremediation. Bioremediation uses microorganisms, microbial or plants enzymes to detoxify contaminants in the soil and other environments. Three group of microorganisms i.e., bacteria, actinomycetes and fungi have been found to degrade the pesticides. Bacterial group involved in degradation includes Bacillus, Pseudomonas, Flavobacterium, Arthrobacter, Paracoccus, Aerobacter, Alcaligens, and Sphingomonas. Fungi having potential bioremediation capacities includes Fusarium, Aspergilus niger, Penicillium, Oxysporum. Streptomycetes have been found to degrade pesticides among the actinomycetes group. This review highlights various aspects of microbial degradation of pesticides including the types of pesticide degrading microbes and mechanism of degradation.