Recent Patents on Biotechnology - Volume 12, Issue 2, 2018

Background: The tremendous need of lipase for varied applications in biotechnology increases its economical value in food and allied industries. Patents suggest that lipase has an impressive number of applications viz. enhancements of flavor in food products (Cheese, butter, alcoholic beverages, milk chocolate and diet control food stuffs), detergent industry in removing oil, grease stain, organic chemical processing, textile industry, oleochemical industry, cosmetic industry and also as therapeutic agents in pharmaceutical industries. Objective: This communication extends the frontier of lipase catalyzed benefits to human body by lowering serum cholesterol and enhancement of flavor in different food products. Methods: Among all, multiple innovations going on in the field of lipase applications are widening its scope in food industries consistently. Therefore, in the present work an effort has been made to explore the utilization of lipase in the field of food product enhancement. Results: Supplementation of food products with lipase results in modification of its physical, chemical and biochemical properties by enhancing its therapeutic activity. Conclusion: Lipases are the most important enzymes used in food industries. They are utilized as industrial catalysts for lipid hydrolysis. Because of lipases hydrolysis nature it is widely exploited to catalyze lipids or fats in different food products and enhancement of food flavors.

Background: Human milk oligosaccharide (HMO) is a third most abundant component in breast milk. HMOs are molecules naturally tailored to the need of an infant. They protect infants from diseases, foster healthy gastrointestinal systems, reinforce fledgling immune function, and promote early brain development. Supplement HMOs to infant formula, which lack this critical element, would substantially improve the function of formula. Overwhelming evidence also indicate that HMOs can be used for the treatment of arthritis and related autoimmune disease, and inhibition of bacteria adhesion or as potential prebiotics. The prospect of using HMO in these applications has stimulated worldwide interest in developing synthesis technology for these valuable products. Methods: As the quantities extracted from human milk are limited, and chemical synthesis methods are time-consuming, costly, and complex, biotechnological approach, involving either enzyme catalysis or metabolically engineered bacteria is preferred. In this review, we highlight the most recent advances in the synthesis technologies, as disclosed in patents and patent applications, and analyze these technologies against those reported in literatures. Conclusion: Significant progress has been made over the past decade, especially in whole-cell biocatalysis for smaller HMOs. Significant challenges remain for larger and more complex HMOs.

Objective: Fully humanized monoclonal antibodies have revolutionized the treatment of many solid tumors, including ovarian, lung, colorectal, and breast cancer. Among the most widely used monoclonal antibodies in clinical oncology are cetuximab, trastuzumab, rituximab, and bevacizumab. Methods: This is a review article focusing on the drug patents for cetuximab, trastuzumab, rituximab, and bevacizumab. Results: These four monoclonal antibodies are used in both first and second line treatment regimens for multiple common malignancies. With recent patent expirations, pharmaceutical companies involved in biosimilar manufacture are looking to establish ownership over these financial monopolies. Conclusion: This article will review these four notable monoclonal antibodies, their role in clinical oncology, and the drug patents that are nearing expiration. There are currently only two biosimilars approved in the United States: the trastuzumab biosimilar trastuzumab-dkst (Ogivri) and the bevacizumab biosimilar bevacizumab-awwb (Mvasi). In Europe, SB3, a trastuzumab biosimilar, is approved for use as well as two rituzimab biosimilars, truxima and rixathon.

Background: Phytases are enzymes capable of degrading phytic acid and are used in animal feed supplementation in order to improve digestibility through the release of minerals such as phosphorus. Recent inventions show interest in production and optimization of recombinant phytases with biochemical and physicochemical characteristics promising for animal feed industry. Methods: This review article is focused on relevant patents of promising phytases, together with the commonly used expression systems for their production and tools currently employed to generate new phytases. We revised all patents related to recombinant phytases and their application in the animal feed industry. The following patents databases were consulted: European Patent Office (Espacenet), the United States Patent and Trademark Office (USPTO), the United States Latin America Patents (LATIPAT), Patent scope -Search International and National Patent Collections (WIPO) and Google Patents. Results: In this review, information was collected from recent publications, including 38 patents related production systems for different recombinant phytases and their application in the animal feed industry. Conclusion: We showed that important recombinant phytases were successfully produced in different expression systems. In addition, this work highlights certain biotechnological tools such as mutagenesis for generation of novel enzymes with biochemical properties of use in the animal feed industry.

Background: Glucose sensors have been extensively researched in patent studies and manufactured a tool for clinical diabetes diagnosis. Although some kinds of electrochemical enzymatic glucose sensors have been commercially successful, there is still room for improvement, in selectivity and reliability of these sensors. Because of the intrinsic disadvantages of enzymes, such as high fabrication cost and poor stability, non-enzymatic glucose sensors have recently been promoted as next generation diagnostic tool due to their relatively low cost, high stability, prompt response, and accuracy. Objective: In this research, a novel free standing and binder free non-enzymatic electrochemical sensor was manufactured using in situ grown copper (Cu) and cobalt (Co) on a silicon (Si) substrate. Methods: Scanning High-Energy Electron Diffraction (SHEED) and Edward deposition methods were used to synthesise the sensors. Results: Morphological observations showed that Cu and Co homogeneously formed nanorod-like shapes over the Si substrate. The elemental composition and structure of the prepared sensors were identified by Reflection High-Energy Electron Diffraction (RHEED). In terms of electrochemical properties, for the enzyme-free glucose sensor, voltammograms showed that the peak currents increased when the glucose solution was injected into the electrolytic cell. The electrical relation of voltage versus current was linear, as shown in the experimental data. Another effective parameter changed the magnetic field; and the linear behaviour of the electrical resistance of Co remained unaltered. Conclusion: In the optimum annealing temperature, where the magnetic field increased, the properties of the samples remained constant. In other words, in the selected annealing temperature, resistance and stability of the layers increased in a significant manner.

Background: The review of literature and patents shows that enhancing the polygalacturonase (PG) production and activity are still required to fulfill the increasing demands. Methods: A dual optimization process, which involved Plackett-Burman design (PBD), with seven factors, and response surface methodology, was applied to optimize the production of extracellular PG enzyme produced by a novel strain of Aspergillus flavus isolated from rotten orange fruit. The fungal PG was purified and biochemically characterized. Results: Three variables (harvesting time, pH and orange pomace concentration), that were verified to be significant by the PBD analysis, were comprehensively optimized via Box-Behnken design. According to this optimization, the highest PG activity (4073 U/mL) was obtained under pH 7 after 48 h using 40 g/L orange pomace as a substrate, with enhancement in PG activity by 51% compared to the first PBD optimization step. The specific activity of the purified PG was 1608 U/mg with polygalacturonic acid and its molecular weight was 55 kDa. The optimum pH was 5 with relative thermal stability (80%) at 50#154;C after 30 min. The PG activity improved in the presence of Cu2+ and Ca2+, while Ba2+, Fe2+ and Zn2+ greatly inhibited the enzyme activity. The obvious Km and Vmax values were 0.8 mg/mL and 2000 μmol/min, respectively. Conclusion: This study is a starting point for initial research in the field of optimization and characterization of A. flavus PG. The statistical optimization of A. flavus PG and its biochemical characterization clearly revealed that this fungal strain can be a potential producer of PG which has a wide range of industrial applications.

Background: The topographical surficial characteristics and properties of materials can induce different cell reactions regarding the viability and adhesion according to the recent patents. The aim of this in vitro experimental study was to evaluate the viability and adhesion of fibroblastic cells seeded on titanium disks with different surface topography. In addition, we revised all patents related to surface treatment of dental implants. Methods: Fibroblast cells were cultured on 6 surface models: mA disks corresponding to titanium surfaces without additional treatment (machined surface) were compared with mB disks, where surfaces were sandblasting etching using aluminum oxide; mC disks where titanium oxide was used for sandblasting etching; mD disks where triple acid conditioning was used; mE disks with laser treated surface; and mF disks which were made of titanium powder by a sintering process. Different surfaces were analyzed using an optical laser profilometer and SEM analysis. To evaluate cell viability on different disc surfaces, a fibroblast cell line derived from monkey kidney (VERO) was in vitro cultured on treated disks surface and cell viability assays were performed to compare growth of fibroblastic cells. Then, the adhesion of the cells was tested by washing procedure. Results: The disks mA, mB, mC and mD less pronounced rugosities were observed and, disc in the mE and mF disks displayed a deeper pronounced surface. The cell viability and adhesion analysis showed significant higher levels on titanium disks surface obtained by the mF method. Conclusion: Our analysis showed that the surface morphology of titanium disc, independent of the roughness parameters, affects cell viability and adhesion differentially, since a higher percentage of viable and attached cells were observed on the mF disks in comparison with the other evaluated surfaces.