Recent Patents on Biotechnology - Current Issue

The increasing industrialization and hydrocarbon use have led to concerning soil contamination. Oil spills and improper disposal of oily waste pose threats to ecosystems and human health. The recovery of these environments is essential, but separating oily components from soil remains challenging. Current bioremediation strategies using synthetic surfactants can cause secondary contamination. Microbial biosurfactants, which are biodegradable and low in toxicity, emerge as promising solutions, and this study reviews methods for utilizing these biosurfactants in the environmental bioremediation of hydrocarbons.

This study explores the efficient and eco-friendly use of biosurfactants for hydrocarbon-contaminated soil management, providing a market-oriented analysis of recent patents and trends, and highlighting the transition from academic research to industrial applications.

The methodology involves an extensive literature review, careful selection of recent studies and patents on biosurfactants in hydrocarbon bioremediation, critical analysis of in-situ and ex-situ application methods, assessment of commercial viability, and synthesis of findings to contribute to sustainable solutions in contaminated environments.

The present study demonstrates the extensive applicability of biosurfactants across various industrial sectors. The increasing interest in incorporating biosurfactants into industrial processes is driven by the pressing need for sustainable solutions to address tangible market challenges. Notably, the cosmetics industry exhibited the highest number of patents related to the use of biosurfactants, underscoring its significant role in advancing the adoption of these environmentally friendly agents. This trend highlights the critical demand for sustainable alternatives in product formulations and underscores the pivotal role of biosurfactants in fostering eco-innovation within the industry.

Biorefineries can refer to forms of fuel production through renewable biomass derivatives, using different structures of lignocellulosic material, such as lignin, hemicellulose, and cellulose. From lignin, we can produce natural binders and adhesives, among other products. With hemicellulose, we can produce emulsifiers, resins, or lubricants, for example. Using cellulose, we can produce fuels, such as ethanol and biodiesel, or even solvents. Fuels from biorefineries can replace, totally or partially, non-renewable fuels that pollute the environment, such as oil. Considering the climate emergency, we are experiencing, the tendency to reduce the availability of oil, and the negative environmental impacts caused by it, fuels obtained through the processing of renewable plant materials present themselves as a good alternative to replacing fossil fuels. First-generation ethanol (1G) can be obtained by fermenting, for example, sugar cane juice. Second-generation (2G) ethanol can be obtained by processing lignocellulosic waste. In this process, there must be pre-treatment and hydrolysis of the biomass before the fermentation and distillation processes. Third-generation ethanol (3G) can be obtained through the fermentation of substrate present in microalgae. Fourth-generation ethanol (4G), in turn, involves the integration of production processes from other generations, increasing the efficiency of 2G and 3G processes for ethanol production.

The objective of this study was to investigate the scenario of patent registrations filed both on the google patents platform and espacenet, which proposes the production of fuels from biorefineries, that are renewable and sustainable.

Although there are other lignocellulosic products originating from biorefineries, we will limit ourselves to patents aimed at the production of cellulosic ethanol. The search covered patents filed in the last 5 years (2019-2023). The 10 patents from each of the 3 biotechnological areas were selected, classified as agriculture, environment, and bioprocesses/bioengineering, totaling 30 patents to be analyzed. After selecting patents through the insertion of keywords and Boolean operators, the patents were selected by reading the title, its summary, and, finally, the full document to verify which were aligned with the study.

Analysis of the documents revealed that, in most cases, China leads the way in patent applications involving the use of fuels, such as cellulosic ethanol, which are environmentally renewable and sustainable. The main strategies for the production of renewable and sustainable fuels in the context of biorefineries explore mechanisms for reusing agricultural waste, pre-treatment of residual biomass, and reuse of biorefinery waste, among other technologies.

The future perspective is that the production of renewable and sustainable energy, such as that coming from biorefinery fuels, has solved its challenges and contributes to the growing global energy transition process. Analyzing and incorporating advances found through patent analysis into technological development provides mechanisms for better performance in the biorefinery sector. In addition to innovations, it is possible to analyze economic and environmental challenges, promoting integrated strategies that combine sustainability and commercial viability. Thus, the energy transition can be accelerated with solutions that increase the efficiency and technological innovation of biorefineries.

The primary objective of this review was to investigate the therapeutic implications of green-synthesized metallic nanoparticles, such as silver and zinc nanoparticles, in addressing dental caries, with a focus on their antibacterial properties and potential to improve current treatment modalities.

In this study, a comprehensive search was conducted across multiple databases, including PubMed, Google Scholar, SID, Scopus, Medline, and Web of Science, as well as selected herbal medicine journals. The inclusion of herbal medicine journals was justified by the growing interest in green-synthesized nanoparticles derived from plant extracts, which are relevant to the focus of this review. Articles published between 2015 and 2023 were evaluated. To ensure a robust selection process, studies were assessed for quality using specific criteria beyond the AXIS checklist, including study design, sample size, and methodological rigor. Any discrepancies between reviewers were resolved through discussion or consultation with a third reviewer. This approach aimed to minimize bias and ensure the inclusion of high-quality studies that contribute meaningfully to the review’s objectives.

In this study, 22 clinical trials were reviewed, and the effectiveness of synthesized nanoparticles was demonstrated in nearly all of them. The nanoparticles found to be most effective against tooth decay-causing bacteria include silver, zinc, selenium, nickel, and copper nanoparticles. To provide greater clarity, key findings, such as nanoparticle size, minimum inhibitory concentration (MIC) values, and specific antibacterial effects, are presented in a table. Additionally, a more in-depth comparison was made between these nanoparticles, focusing on their relative effectiveness under different conditions. For example, silver nanoparticles were consistently effective across various trials due to their strong antimicrobial properties, while zinc and copper nanoparticles showed efficacy in specific concentrations and against particular bacterial strains. Furthermore, green-synthesized nanoparticles were compared with those produced using traditional methods, evaluating factors, such as effectiveness, scalability, and safety, to provide a comprehensive understanding of their potential in dental caries treatment.

According to the literature review, nanoparticles with antibacterial properties have the potential to serve as an alternative or complement to conventional chemical treatments for combating tooth decay. Future research should focus on exploring specific types of nanoparticles, such as green-synthesized metallic nanoparticles (e.g., silver and zinc), which have shown promising antibacterial effects, Many of these ideas are patentable. Additionally, more emphasis should be placed on optimizing synthesis methods that enhance biocompatibility and scalability for clinical use. The development of affordable treatment options using medicinal plants for green synthesis is particularly promising and should be further explored to make nanoparticle-based therapies more accessible in clinical settings.

The present study examined Polyhydroxy butyrate production (PHB) potential of different photosynthetic microbes such as Chlorella vulgaris, Scenedesmus obliquus and Rhodobacter capsulatus-PK under different nutrient conditions. Biodegradable bioplastics, such as Poly-β-hydroxybutyrates (PHB), derived from these microbes provide a sustainable alternative to conventional petroleum-based non-degradable plastics.

As the demand for clean and sustainable alternatives rises, bio-plastic is gaining attention as a viable substitute to conventional plastics. However, conventional sources of bio-plastic production have inherent limitations, which can be effectively addressed through the utilization of photosynthetic microbes e.g., microalgae, purple non sulphur bacteria.

The production of bioplastic was evaluated by cultivating the microalgae in BG-11, BBM and PNSB in synthetic growth media (MI, MII) with different nitrogen concentrations of 0%, 50% and 100%. The biopolymer (PHB) was obtained from all experiments in a wide range of concentration (7-42.8%) of dry cell weight (DCW).

In this study, algal isolate SK1 demonstrated the highest PHB content (42.8%) in BBM under 100% nitrogen starvations rendering the bioplastic exceptionally compatible and suitable for eco-friendly applications. Additionally, various patents cited by different authors on different aspects of microbial bioplastic production.

Nutrition depletion such as nitrogen scarcity induced stressful growth conditions that resulted in highest accumulation of the biopolymer PHB. Optimizing nitrogen availability is key to maximizing PHB production, making it a viable sustainable alternative to conventional plastics.

One of the main sources of contaminated dairy products is moldy fungi, specifically species of Aspergillus and Penicillium. This study aimed to evaluate the effect of the alcoholic extract of Allium jesdianum plant on the growth of molds contaminating dairy products in Isfahan.

In this patent research, 200 samples of dairy products were gathered from different areas of Isfahan city, including 70 samples of cheese, 60 samples of buttermilk, 40 samples of yogurt, 20 samples of curd, and 10 samples of cream. The antifungal activity of Allium jesdianum plant was investigated by the diffusion method in disc, well, and agar dilution in various concentrations. Minimum inhibitory concentration (MIC) and minimum fatal concentration (MFC) were also determined. Aspergillus, Penicillium, Cladosporium, and Acremonium fungi were the most commonly found fungal contaminants of this investigation. Antifungal activity was not observed by disc diffusion and well diffusion methods.

In the agar dilution method, ethanolic and methanolic extracts of stem and leaves in concentrations of 80, 60, 40, and 30 mg/ml, and ethanolic and methanolic extracts of plant bulbs in concentrations of 60 and 30 mg/ml revealed antifungal activity against Aspergillus niger, Penicillium notatum, and Penicillium chrysogenum. The MIC of stem and leaf ethanol extracts and onion ethanol for Aspergillus niger was 18.7, and for Penicillium notatum and Penicillium chrysogenum, it was 37.5, 37.5, 37.5, and 37.5 mg/ml. Fungal contamination of dairy products is a serious threat to the public health of society. Therefore, identifying medicinal plants with antifungal activity can be an effective step in preventing fungal contamination and increasing the shelf life of these products.

The results of this research have shown that the Allium jesdianum plant can inhibit the growth of Aspergillus niger, Penicillium notatum, and Penicillium chrysogenum.

Among the bioactive isoflavones identified from different plants is formononetin. Formononetin's antioxidant, anti-inflammatory, and anti-cancer qualities have all received a lot of attention lately. The goal of the current investigation was to examine formononetin's antifungal and antibacterial activity against Candida albicans and Enterococcus faecalis in vitro, respectively.

The present study determined the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and Minimum Fungicidal Concentration (MFC) of formononetin, chlorhexidine, formononetin + chlorhexidine, Formononetin + nystatin, and nystatin against E. faecalis and C. albicans isolates. Next, SPSS version 25 was used to analyze the data. A significance cutoff of P<0.05 was taken into account.

Chlorhexidine + formononetin and formononetin alone had the greatest and lowest minimum inhibitory concentrations (MIC) against E. faecalis, at 6.6 and 18.3 μg/ml, respectively. With MIC values of 0.66 and 8.3 μg/ml against a C. albicans isolate sensitive to nystatin, the combination of formononetin + nystatin and formononetin alone was shown to have the greatest and lowest anti-candida effects.

The findings demonstrated a considerably greater antibacterial effect (P<0.05) for the combinations of formononetin + nystatin and chlorhexidine as compared to nystatin and chlorhexidine alone.

Bacillus amyloliquefaciens contains several fungal inhibitory compounds, such as peptides and lipopeptides, representing the remarkable potential for biotechnological, agricultural, and biopharmaceutical applications.

This research aimed to extract and characterize iturin A as the key antagonism factor of Bacillus amyloliquefaciens M13RW01 toward pathogenic fungi, using HPLC and mass spectrometry (MS) analysis.

For this study, Bacillus amyloliquefaciens strain M13-RW01 isolated from Isfahan soil was used. The lipopeptide compounds of B. amyloliquefaciens were examined for antagonistic performance against Aspergillus niger PTCC 5010, Mucor hiemalis PTCC 5292, Fusarium oxysporum CBS 62087, and Penicillium chrysogenum PTCC 5037 by well diffusion and percentage of growth inhibition. The crude extract was run on Waters μBondpak C18 column in the HPLC system to separate the antibiotics. Major antibiotics were analyzed based on MS.

HPLC analysis demonstrated that the lipopeptide compound is similar to iturin A. Moreover, MS analysis of these compounds and purified iturin A revealed a high similarity between them, with the same molecular ion peaks identified. Results showed that the produced lipopeptides by Bacillus amyloliquefaciens were of iturin A genum. The molecular ion peaks of the B. amyloliquefaciens M13RW01 methanolic fraction were at 1027.10, 1043.05, 1058, 1066, 1072, 1088.95. These compounds restrained fungal germination and growth. Inhibition growth percentages were 79.28, 76.13, 84.47and 59.15% for Aspergillus niger, Mucor hiemalis, Fusarium oxysporum, and Penicillium chrysogenum, respectively.

According to the present study, B. amyloliquefaciens M13RW01 lipopeptides are able to inhibit the growth of some fungi. B. amyloliquefaciens M13-RW01 isolated from Isfahan soil plays an essential part in antagonizing pathogenic fungi. Thus, this antifungal lipopeptide is supposed to be a biological protection agent for agricultural products and patents.

Trophoblast Cell Surface Antigen 2 (Trop2) is a transmembrane glycoprotein that has been implicated in the progression and metastasis of various cancers, including hepatocellular carcinoma (HCC). Targeting Trop2 expression may represent a promising approach for the development of novel therapeutic strategies and patents.

This study aimed to investigate the effects of Trop2 knockdown using small interfering RNA (siRNA) on the phenotypic and molecular characteristics of the HepG2 liver cancer cell line.

HepG2 cells were transfected with different concentrations of Trop2-targeting siRNA (3 nM, 5 nM, and 7 nM) at various time intervals (6, 24, and 48 hrs). The expression of Trop2 was assessed by real-time PCR before and after transfection. The impact of Trop2 knockdown on cell apoptosis, migration, morphology, histopathological features, wound-healing assays, and microscopic analysis was examined. Additionally, the expression of the TPM1 gene was evaluated using immunohistochemical analysis.

Trop2 mRNA level was significantly decreased in HepG2 cells in a time- and concentration-dependent manner following siRNA transfection. The downregulation of Trop2 resulted in a marked increase in apoptosis, a reduction in cell migration, and alterations in cell morphology and histopathological characteristics. Furthermore, the expression of the TPM1 gene was found to be upregulated in Trop2-knockdown HepG2 cells.

These results highlight the potential of Trop2 as a therapeutic target for the management of hepatocellular carcinoma.

Salinity is one of the primary environmental factors that significantly impact global crop production. Plant growth promoting rhizobacteria (PGPR) inoculation to crops improves the productivity of the crops.

To develop a biofertilizer specifically for saline soil, bacteria were isolated from the rhizosphere of mustard plants along with the plant growth-promoting traits grown in saline soil (EC 6 dS m). Halotolerant 22 bacterial strains were isolated and identified from the rhizospheric soil mustard crop, Purvanchal (Indian state). According to the study, 54.54% of the isolates had phosphate solubilization efficiencies ranging from 7% to 27% on plate assays. According to quantitative measurements, 63.63% of the strains exhibited the ability to solubilize phosphate, with degrees of solubilization varying between 0.49 and 3.34 µg/ml. Furthermore, 50% of the isolates showed the ability to solubilize zinc, with solubilization rates varying from 12% to 53%. Further 59.09% of the bacterial strains showed ammonium production test; these strains were classified as having low (+), medium (++), and high (+++) levels of ammonium production.

According to the research, these halo-tolerant plant growth-promoting rhizobacteria (PGPR) have particular functional properties that may help mustard crops grow more rapidly in salinity-stressed environments. Because these PGPR strains increase nutrient availability and stimulate plant development, they may find use in agriculture, especially in saline settings.

The study emphasizes how crucial it is to use PGPR with particular nutrient mobilization features to promote crop growth under difficult circumstances. The identification of these efficient strains may lead to the development of patent biofertilizers designed for saline soils, further supporting their application in modern agricultural practices.

This study aims to develop an efficient and reproducible in vitro protocol for high-frequency embryogenic callus induction and subsequent plant regeneration in multiple sorghum (Sorghum bicolor L. Moench) cultivars, thereby establishing a foundation for genetic transformation, mutation breeding, and other biotechnological applications aimed at enhancing sorghum crop improvement and productivity.

Sorghum (Sorghum bicolor (L.) Moench) is an important cereal crop known for its adaptability to harsh environments and nutritional value. Despite its significance, sorghum remains challenging for in vitro propagation due to difficulties in regenerating callus tissue, especially from monocotyledonous explants. Callus induction and regeneration protocols are crucial for genetic transformation, mutation breeding, and biotechnological applications in sorghum improvement.

To establish an effective in vitro protocol for callus induction and subsequent plant regeneration using different sorghum cultivars, optimizing conditions for high-frequency embryogenic callus formation and plant regeneration.

Six sorghum cultivars (IS 3477, IS 33095, IS 7155, IS 2898, IS 7005, and IS 1202) were selected. Immature inflorescence explants were cultured on a modified Murashige and Skoog's (MS) medium with 3% sucrose, 0.8% agar, and 2.0 mg/l 2,4-D for callus induction. After 14 days, embryogenic and non-embryogenic calli were distinguished. Regeneration media were optimized using embryogenic calli, with 1.5 mg/l 6-benzylaminopurine (BAP) for shoot development and 1 mg/l NAA (1-naphthaleneacetic acid) in a half-strength MS medium for root development.

Two distinct forms of calli were observed: a non-embryogenic light yellow callus and a white, granular embryogenic callus. Embryogenic callus induction frequency varied from 40% to 96% among the cultivars, with IS 3477 and IS 33095 exhibiting the highest frequencies (96% and 88%, respectively), while IS 1202 showed the lowest (40%). Regenerated shoots were successfully developed within 6-18 days and later transferred to a rooting medium, resulting in healthy plantlets. Transplanted plantlets showed normal growth and no morphological abnormalities in the field.

This study provides a reliable protocol for efficient callus induction and plant regeneration in multiple sorghum cultivars. The optimized conditions can be utilized for genetic studies, crop improvement, and biotechnological applications, thus contributing to the advancement of sorghum breeding and biotechnology research.