Recent Patents on Biotechnology - Volume 15, Issue 3, 2021

Extremophilic microorganisms from a wide variety of extreme natural environments have been researched, and many biotechnological applications have been carried out, due to their capacity to produce biomolecules resistant to extreme conditions, such as fibrinolytic proteases. The search for new fibrinolytic enzymes is important in the development of new therapies against cardiovascular diseases. This article aimed to evaluate the patents filed about protease with fibrinolytic activity produced by extremophilic microorganisms whose use is aimed at the development of new drugs for the treatment of cardiovascular diseases. The prospecting was carried out using data on deposits and patent concessions made available on the technological bases: European Patent Office (EPO), United States Patent and Trademark Office (USPTO), World Intellectual Property Organization (WIPO), Instituto Nacional de Propriedade Industrial – Brazil (INPI), The LENS and Patent Inspiration. The International Patent Classification and subclasses and groups for each document were also evaluated. Although 382 patents were selected using terms related to extreme environments, such as “thermophile” and “acidophiles”, few were related to clinical use and were mainly performed using Bacillus subtilis and Streptomyces megasporus strains. A highlight of nattokinase was produced by Bacillus subtilis GDN and actinokinase by Streptomyces megasporus SD5. The low number of patents on enzymes with this profile (extreme environments) revealed a little-explored field, promising in the development of new microbial thrombolytic drugs, such as fibrinolytic enzymes with less adverse effects.

Plants of the genus Cannabis have been gaining attention in medicine. The most outstanding treatments include antispasmodic, analgesic, antiemetic, neuroprotective and anti-inflammatory actions. The aim of the study was to describe patents associated with cannabis and cannabinoids as a treatment for pregnant women. A search was carried out on four patent bases: National Institute of Industrial Property, Latipat, Espacenet and Patentscope. The used search keys were “Cannabis”, “Cannabidiol”, “Cannabinoids”, “CBD” and “THC” associated with “pregnant women”. Patents in any language and year were considered. A total of 1,402 patents were found in all the used databases. After reading the title and abstract, patents were excluded if they did not meet the eligibility criteria and/or if they were duplicates, thus leaving 11 patents on the use of cannabis and/or cannabinoids as treatment in pregnant women. A critical analysis was carried out. All selected patents were from the Asian continent (China), focusing on traditional Chinese medicine for treating various possible changes during pregnancy and also for diseases prevention in the postpartum period. It was found that the main causes for the need of such treatments were related to spleen deficiencies, inability and/or moisture stagnation. Finally, the vast majority of treatments used Cannabis fruits associated with other plants. Given the above, patents related to cannabis and cannabinoids for the treatment of pregnant women in the peri and postnatal periods were found to be associated with traditional Chinese medicine.

Background: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure. Objectives: The aim of the present study was to evaluate the multiplex polymerase chain reaction (PCR) for detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children with hospital- acquired sepsis compared to the conventional biochemical reactions for identification of these organisms. Methods: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR with primers specific to the 3 tested bacteria. Results: Multiplex PCR was positive in 96% of isolates and 4 isolates had negative results. Falsepositive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa and 27 isolates of Stenotrophomonas maltophilia. The diagnostic value of the multiplex PCR compared to the biochemical identification revealed sensitivity 96.04%, specificity 96.9%, positive predictive value 97.00%, negative predictive value 96.00% and accuracy 96.50%. Conclusion: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific and accurate. The accuracy differs according to the organisms with 100% accuracy for Acinetobacter baumannii.

Background: 1,8 cineole-rich supercritical CO2 extract of small cardamom seeds of Alleppey green variety exhibiting prominent antioxidant property was microencapsulated and utilized in formulating an antioxidant-rich custard. However, the antioxidant potency of the prepared custard was not appreciable. To redress the phytochemical loss during custard preparation, custard using nanoliposomes was formulated. Patents related to 1,8 cineole-rich food products have been revised thoroughly. Objective: The objective of the current study is to examine whether nanoencapsulationmediated entrapment of antioxidants is more effective in fortifying a dessert, namely custard, vis-à-vis microencapsulated (spray dried)-mediated enhancement of antioxidative potency in the same. Methods: Our previous investigations have established that nanoliposome of 1,8 cineole- rich supercritical CO2 extract of small cardamom seeds effectively redresses type 2 diabetes and hypercholesterolemia. In the current investigation, this pre-characterized nanoliposome which exhibited appreciable in vitro and in vivo antioxidant efficacy has been utilized at varying concentrations for fortification of a custard. The designer custard samples have been characterized for their sensory and physicochemical properties, identification of the cardamom antioxidants therein and determination of the synergistic efficacy value of the identified antioxidants. Results: The custard formulated with 0.3% nanoliposomes exhibited appreciable antioxidant potency in terms of DPPH radical scavenging activity (304.58±1.09 mg/ml) and reducing power (0.020±0.001 mg BHT/g custard), conferred by its total phenolic content (0.049±0.004 mg GAE/g custard). It also had relatively more stable textural attributes vis-à-vis the control sample (formulated with the non-encapsulated native extract). GCMS analysis of the nanoliposome-fortified custard confirmed retention of the spice antioxidants, namely1,8- cineole, α-terpinyl acetate, α-terpineol and linalool and its synergistic efficacy value being greater than unity, attested to the synergistic presence of the said antioxidants therein. The newly formulated custard retained more than 4.5 times of 1,8-cineole (5.05 mg/g custard) vis-à-vis the custard sample (1.12 mg/g custard) prepared with a microencapsulated (spray-dried) formulation of the extract. Additionally, the absence of heavy metals in the formulated custard confirmed it to be safe for human consumption. Conclusion: This is the first study on the application of nanoliposomes of spiceuticals in the formulation of a dessert, and more emphatically on use of a ‘green’ supercritical CO2 extract of spice antioxidants in fortification of a dessert to achieve antioxidant synergy.

Background: Carotenoids are known as lipophilic secondary metabolites with important biological activities, which are mostly used in the food and pharmaceutical industry. They contribute to the colours of many fruits and flowers. Studies on the biosynthetic pathways of isoprenoids and carotenoids are still scarce, especially in microalgae and are limited to specic groups. Dunaliella spp. in Chlorophyta taxon of algae, the 2-C-methyl-D-erythritol 4-phosphate/ 1-deoxy-D-xylulose 5-phosphate (DOXP/MEP) is the synthesis pathway of sterols and carotenoids. Objectives: In this study, we used 12 Psy gene sequences in Dunaliella sp., also Scenedesmus acutus and Diospyros kaki to investigate a genome-wide search. The results are useful for better identification of carotenoids metabolisms, and increasing the production rate of beta-carotene in pharmaceutical, food and industrial processes. Methods: Phytoene synthase (Psy) from Dunaliella spp. was selected as the rst regulatory point in the carotenoids pathway that catalysis the formation of geranylgeranyl pyrophosphate in isoprenoid biosynthesis. Structural, evolutionary and physics-chemical characteristics were investigated, using various bioinformatics tools and computer techniques. Moreover, some recently published patents were also regarded. Results: The maximum length of the conserved motif was 5167 bp for Dunaliella. sp. (DQ463306.1) and the smallest length of the conserved motif was 416 bp belonging to D. salina (JQ762451.1). The average molecular weight of species was 41820.53 Da. The theoretical pI of species varied from 4.87 to 9.65, indicating vernation in the acidic nature. Two strains of D. bardawil (U91900.1 and EU328287.1) showed just a long-distance relationship with all other Dunaliella strains. Whilst, D. parva displayed the furthest vicinity with all the studied strains. Conclusion: Our study highlighted the Psy regulatory mechanism, as a key factor in the carotenoids pathway, to facilitate genetic and metabolic engineering studies. The obtained tree-dimensional arrangement of the amino acids revealed the regional structures and folding of the diverse segments of helices, sheets and turns. This information is a key point to unveil the protein's operation mechanism. Besides, we confirmed the suitability of bioinformatic approaches for analysing the gene structures and identifying the new Psy genes in unstudied microalgal strains.

Background: Aedes aegypti mosquitoes transmit dengue, zika, and chikungunya viruses, neglected diseases that are considered global health challenges. Due to the lack of antiviral drugs and vaccines for these illnesses, vector control with chemical insecticides is the principal strategy for preventing their spread. However, vector populations are becoming increasingly resistant to insecticides, and the development of other control measures is, therefore, imperative. Methods: A new insect trap (IT) was used to control Aedes aegypti. A specific light-emitting diode (LED) served as the attractant based on specific wavelength ranges (450-495, 500-550 and 570-600 nm). The IT utilized insect-attracting and killing mechanisms that included a black capture box, a suction-producing mechanism, an electric shock device and a nylon cloth device that held surviving mosquitoes, which died due to starvation. Capture assays of twenty non-feeding females inside a cage were performed in triplicate using different LED intensities. A commercial trap (ultraviolet lamp attractant and suction system) was used as a positive control. Results: Capture assays of A. aegypti with different intensities and LED combinations showed that the tricolored trap captured 100% of the females, followed by the Green LED 8 set, which captured 91%; in comparison, commercial traps captured approximately 25% of the insects. Although there were no significant differences between the experimental groups, the tricolored trap probably will capture more mosquito females considering the vision variation in individual females. Conclusion: We herein present a green technology-based IT that is effective, safe and successful for reducing mosquito populations, thereby preventing mosquito-borne disease spread.

Background: Starter cultures are essential in food industry biotechnology, consisting of microorganism preparations inoculated to produce safe fermented foods with desirable sensory characteristics. Objective: This study aims to identify and analyze the growth and flow of knowledge regarding starter cultures by creating scientific and technological profiles using patentometric and bibliometric indicators. Methods: A search for patents and scientific articles was conducted in December 2020 following a proposed 10-step methodology using the Scopus® and Patentinspiration databases. The search strategy was based on the keywords “starter culture” and “fermentation” considering publications up to 2020. Results: A total of 3035 articles and 719 patents were published until 2020, presenting a more significant number in the last ten (10) years due to the development of biological sciences and molecular biology involving enzymes and microorganisms. Italy leads the scientific production while China leads the technological. It was also possible to determine the most productive author and inventors, the most influential articles and inventions, and the main scientific journals and patent offices. Conclusion: Scientific and technological activities have an exponential behavior showing that the knowledge about starter cultures continues to grow, becoming a field of interest for optimizing industrial processes related to food fermentation, thus achieving diversification of products that can satisfy the demand for food in an increasingly competitive global market.