Recent Patents on Biotechnology - Online First

Biodiesel from animal waste provides an alternative source of fuel. It is eco-friendly and cheaper than conventional fuel obtained through the distillation of crude oil. Biodiesel is similar to petroleum diesel and can be used alone or blended with fossil diesel as an energy source. This study provides insight into the use of waste animal fats for biodiesel production.

The data were retrieved from different sources, including PubMed, Google Scholar, and other scientific websites. Scope of the study can be clearly understood from the facts revealed from patent work.

The use of waste animal fats provides a renewable and eco-friendly alternative to petroleum diesel. Both homogeneous and heterogeneous catalysis methods have proven effective for processing WAFs, with KOH and NaOH commonly used in concentrations ranging from 1% to 2%. Increasing the catalyst concentration within a certain range can enhance biodiesel yield. However, the use of acid catalysts for transesterification of WAFs is time-intensive and requires a high alcohol-to-fat ratio. Despite these advantages, challenges remain in catalyst reuse for certain reactions and in managing complex selectivity combinations. Lipases in water-poor environments are also employed for transesterification.

The fatty acid profiles of both animal and vegetable sources have been found suitable for biodiesel production. Although it is a cheap source for generating fuel, using specific reaction media such as acyl acceptors and employing a combination of two enzymes on a specialized support can help further reduce costs.

Currently, biodiesel is widely used in developed countries such as the US and Germany, but its adoption remains limited in developing nations. Optimizing transesterification methods—including the use of heterogeneous or homogeneous catalysis—and incorporating innovative technologies such as microwave-assisted enzymes can further enhance biodiesel yield and efficiency.

Nanoemulsions (NEs) are a biphasic systems formed by two immiscible liquids: (1) oil-in-water or (2) water-in-oil emulsions. These systems exhibit unique physicochemical properties due to their small particle size, making them more versatile compared to other emulsion systems. Thus, this research aimed to produce and evaluate oil-in-water nanoemulsions.

NE’s were prepared using Squalene, grape seed, and avocado oils, combined with soy lecithin or soy phosphatidylcholine as emulsifiers. The selection of oils considered both regulatory aspects and patent restrictions. Formulations were produced by microfluidization, and their physicochemical properties, such as particle size, polydispersity index, and zeta potential, were assessed at day one (D0) and day 7 (D7), under storage at 4°C, 25°C, and 40°C.

The nanoemulsions produced with soy phosphatidylcholine and vegetable oils exhibited good preliminary stability, due to their small particle size. In contrast, formulations using soy lecithin as the emulsifier did not show favorable results, especially the one with grape seed oil, which showed a large particle diameter. However, none of these formulations exhibited cellular cytotoxicity.

Emulsifier selection had a strong impact on NEs characteristics, with Soy-PC producing smaller and more uniform particles compared to soy lecithin. All NEs showed good biocompatibility in fibroblasts, indicating their safety.

Among the formulations, the one with phosphatidylcholine demonstrated a safer and more reliable stability profile, making it a promising candidate for application in the pharmaceutical and cosmetics industries.

Urinary Tract Infections (UTIs) caused by Escherichia coli are a major concern due to rising antimicrobial resistance. Long non-coding RNAs (lncRNAs) play crucial roles in gene regulation, but their involvement in UTIs and their implication in antimicrobial resistance mechanisms are not well understood. This study investigates the association between specific long non-coding RNAs, immune response, and antibiotic resistance in patients with infections caused by E. coli.

Quantification of specific lncRNAs (NEAT1-1, NEAT1-2, MIR3142HG, AK170409, and IL7AS) was performed using quantitative PCR. Inflammatory markers IL-1β, IRF3, and NF-κB were measured in UTI patients using ELISA kits to assess their biological response. The minimum inhibitory concentration of eleven antibiotics was determined by testing all 25 urine samples and categorized as sensitive, intermediate, or resistant.

NEAT1-1, NEAT1-2, MIR3142HG, and AK170409 were significantly upregulated (p < 0.05). Inflammatory markers were significantly elevated in all samples: IL-1β at 72.36±13.8pg/mL, IRF3 at 79.36±15.01pg/mL, and NF-κB at 4.43±0.82pg/mL

(p < 0.0001). NEAT1-2 and AK170409 expression correlated with distinct antibiotic response patterns (p < 0.05). Differences in biological and hematological data were observed among UTI patients with varying topographic expressions of specific long non-coding RNAs (lncRNAs).

Specific lncRNAs may be involved in modulating immune responses and influencing antibiotic susceptibility in UTIs. Their expression patterns reflect both the severity of infection and resistance profiles, suggesting a functional role in the development of antimicrobial resistance.

LncRNAs are potential biomarkers for UTIs caused by E. coli. Future studies should focus on elucidating their role in the development of antibiotic resistance and exploring their patentable applications in biotechnology.

Polycyclic aromatic hydrocarbons (PAHs) are toxic petroleum byproducts in soil, exhibiting significant genotoxic properties. Microorganisms residing in contaminated soils serve as effective detoxifying agents. Among various strategies, bioremediation is an efficient biological method for detoxifying PAHs.

Hundreds of soil samples were collected from the Gulf of Suez, Egypt. The isolation process utilized an enrichment culture system with phenol naphthalene (PN) (10 mg/mL) as the primary carbon source. HPLC analysis was applied to confirm PN degradation. Consequently, the bacterial strain was characterized morphologically, biochemically, and through partial sequencing of its 16S rRNA gene. Subsequently, its plasmid was purified to transfer its phenotype to Escherichia coli. Finally, a bioremediation approach was conducted to test its PAH degradation.

HPLC analysis was performed to confirm PN degradation by the isolated strain. The isolated strain was identified as Lysinibacillus species AAS1 (OR044755.1) with 98.43% sequence similarity to the Lysinibacillus genus. Subsequently, E. coli transformants with the isolated plasmid were grown in the presence of PN as the primary carbon source. Finally, the bioremediation assay of the isolated strain exhibited a high efficiency in detoxifying PN.

The novel identified Lysinibacillus species AAS1 (OR044755.1) shows promise for PAHs detoxification, which may lead to the exploration of a biological agent for the remediation of water, soil contaminated with PAHs and patents.

A novel bacterial strain bearing a plasmid that can degrade PN was isolated from Egyptian petroleum waste-contaminated soil. It paved the way for further studies to isolate the whole gene(s) responsible for such degradation.

The horseshoe crab, a Xiphosurid species with an ancient lineage that dates back 450 million years, has proven to be a precious asset to the pharmaceutical industry. The blood extracted from these creatures is an irreplaceable component in detecting bacterial endotoxins, crucially important in pharmaceutical and functional settings. Unfortunately, these living ancient organisms are threatened from multiple perspectives and are now considered endangered. While efforts to conserve these creatures are underway, exploring technologies for their conservation can help us understand the latest advancements in the field and shed light on areas that have not yet been targeted.

This analytical report is the first of its kind in this domain and provides a comprehensive overview of the available patents associated with the conservation of horseshoe crabs. Patents associated with horseshoe crabs were searched in PatSeer and the data analysed and filtered, based on relevance.

The analysis is based on an extensive dataset (413) that describes technology for conservation of these living fossils, with a focus on recombinant proteins that can be a viable alternative to the mass utilization of the horseshoe crabs for the extraction of limulus amoebocyte lysate. Other technological advances which advocate cell-free hemolymph production and the use of artificial baits to replace the traditional grassroot practices, procedures related to efficient breeding, growth, hatching and release from artificial culture systems can go a long way in the conservation of these living fossils.

The technologies and innovation reveal possible means of reducing the dependence on live animals through non-invasive methods. Novel interventions such as recombinant Factor C for endotoxin detection provide promising alternatives to conventional methods. Additionally, technological advances in aquaculture protocols provide strategies that allow for the conservation and artificial breeding of the horseshoe crab.

Transitioning to recombinant Factor C, a ban on horseshoe crab baits, diversity mapping through genetic markers and artificial breeding techniques are some of the measures that can be manifested at the policy level to enhance conservation efforts.

Aeromonas hydrophila, a rod-shaped, gram-negative bacterium, is frequently found in aquatic surroundings and additionally present in drinking water, sewage, and food sources. This microbe is gaining recognition as a potential threat to health, classifying it as an emerging pathogen. Biosurfactants are microbial-derived compounds that share hydrophilic and hydrophobic moieties that are surface active. This study aimed to investigate the effect of biosurfactant isolated from Actinobacteria on the expression of the bfp gene of A. hydrophila isolated from children's stool samples to 
patent the ideal method in Qom, Iran, from May 2022 to March 2023.

Actinobacteria were isolated from soil samples of the desert areas of Qom province, Iran. Biochemical and molecular tests of 16S rRNA were used to identify Actinobacteria isolates. The produced biosurfactant was investigated by methods of hemolysis, oil droplet destruction, lipase production, oil expansion, emulsifying activity, and surface tension reduction measurement. The structure of biosurfactant was investigated by Fourier transform infrared spectroscopy (FTIR) analysis, and its effect on bfp gene expression was measured. Also, isolates of A. hydrophila were obtained from stool samples of children referred to Hazrat Masoumeh Hospital in Qom from May 2022 to March 2023. Then, the effect of a biosurfactant isolated from Actinobacteria on the bfp gene expression of A. hydrophila isolates was measured by RT-PCR.

Based on sequencing data, the Streptomyces genus with the ability to produce biosurfactant was isolated from the soil of the studied area, which could reduce the expression of the bfp gene after treatment with biosurfactant in clinical isolates of A. hydrophila.

The biosurfactant-producing isolates were identified as Streptomyces spp. The results indicated that the biosurfactant significantly decreased bfp gene expression in A. hydrophila. This emphasizes the potential of biosurfactants to eliminate microorganisms by reducing virulence gene expression, inhibiting biofilm formation, demonstrating antimicrobial activity, and improving emulsification. The study supports the idea that biosurfactants can interfere with bacterial mechanisms that cause disease, such as biofilm formation, which is critical for pathogen persistence and resistance. Previous research also confirms the antipathogenic activity of natural isolates against A. hydrophila.

The findings of the present study show that the desert soils of Qom province are a potential area for finding actinobacterial isolates with the ability to produce biosurfactants and influence the expression of pathogenic genes of clinical isolates of A. hydrophila.