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Bioprospecting Extremophiles for Sustainable Biobased Industry
Published in Pratibha Dheeran, Sachin Kumar, Extremophiles, 2022
Neha Basotra, rashika Raheja, Gaurav Sharma, Kumud Ashish Singh, Diksha Sharma, Rohit Rai, Bhupinder Singh Chadha
Several biorefineries using bioconversion platform for lignocellulosic biomass conversion to bioethanol involves the heterologous expression of cellulolytic enzymes in the Pichia pastoris using glycosylphosphatidylinositol inositol anchor. With the advancement in research recently, combined strategy for cell surface display of cellulolytic genes has been used where the endoglucanase (EG) and cellobiohydrolase I (CBHI) were produced in a β-glucosidase displaying S. cerevisiae (Inokuma et al. 2014). It was observed that display of gene cassette EG-D-CBHI-D and EG-SCBHI-S resulted in higher ethanol production (2.9 and 2.6 g/L from 10 g/L phosphoric acid swollen cellulose, respectively) (Liu et al. 2015). Mannans, which consist of major fraction of hemicelluloses, are also the source of carbohydrates which can be exploited for its subsequent conversion to ethanol. So the use of cell surface display helps the co-display of β-mannanase and β-mannosidase on the yeast cell surface, which has resulted in efficient hydrolysis of recalcitrant lignocellulosiscs (Inokuma et al. 2014, Ishii et al. 2016).
Active Nanoparticle Targeting: Current Status and Future Challenges
Published in Sandeep Nema, John D. Ludwig, Parenteral Medications, 2019
Siddharth Patel, Janni Mirosevich
The clathrin- and caveolae-independent pathways have been demonstrated to shuttle a range of biological materials including viruses (SV40), and proteins (cholera toxin B, glycosylphosphatidylinositol-linked proteins, major histocompatibility complex class 1 (MHC-1) protein, and interleukin) (Doherty and McMahon 2009). Depending on which GTPases are involved, the clathrin- and caveolae-independent pathway can be further categorized into Arf6-dependent, flotillin-dependent, Cdc42-dependent, and RhoA-dependent mechanisms (Doherty and McMahon 2009). Currently, our knowledge and understanding of the clathrin- and caveolae-independent pathways is limited; however, folate-modified nanoparticles and PLGA nanoparticles have been observed to use this pathway (Lu and Low 2002; Qaddoumi et al. 2003).
Extracellular Vesicles (EVs)
Published in Peixuan Guo, Kirill A. Afonin, RNA Nanotechnology and Therapeutics, 2022
Alice Braga, Giulia Manferrari, Jayden A. Smith, Stefano Pluchino
Similar to antibodies, nanobodies, small single-variable domains obtained from heavy-chain antibodies from Camelidae have been used to endow EVs with targeting capabilities. For instance, such structures have been used to target the epidermal growth factor receptor (EGFR) (Kooijmans, Aleza, et al. 2016b), overexpressed in many tumors. The EGa1 nanobody was used due to its high affinity for EGFR and its ability to competitively inhibit natural ligand binding, preventing receptor activation. To facilitate membrane binding of EGa1, fusion to a C-terminal glycosylphosphatidylinositol (GPI) signal peptide from human decay-accelerating factor (DAF) was used. EVs are enriched in lipid raft-associated lipids that include GPI-anchored proteins, with GPI-anchored DAF previously described to be secreted in EVs; thus, human DAF-derived GPI-anchor signal peptide was fused to the nanobodies, and its expression was shown to otherwise not affect EV characteristics. To investigate the efficacy of such-functionalized EVs, derived from Neuro2A mouse neuroblastoma cells, a far-red fluorescent dye was used to track their uptake by recipient Neuro2A, HeLa, or A431 cells. These lines vary in EGFR expression, with levels being undetectable in Neuro2A cells but greatly overexpressed in A431 cells. Concomitantly, incubation with EVs expressing DAF and EGa1 resulted in a significant greater binding to A431 cells, but the uptake of the EVs did not significantly differ from non-functionalized control EVs or among cell types. This unexpected result highlights the need for further exploration of the uptake mechanisms underlying EV-based cargo delivery.
Assessment of environmental and occupational exposure while working with multidrug resistant (MDR) fungus Candida auris in an animal facility
Published in Journal of Occupational and Environmental Hygiene, 2019
Steven R. Torres, Heather C. Kim, Lynn Leach, Sudha Chaturvedi, Corey J. Bennett, David J. Hill, Magdia De Jesus
The current animal models of disseminated C. auris infection use concentrations that range from 106 to 108 cells, along with the immunosuppressive agent cyclophosphamide. These animal models are being used to test new alternative therapeutics, including a small molecule antifungal agent called APX001A derived from the pro-drug APX001 that inhibits the fungal protein Gwt1 (glycosylphosphatidylinositol-anchored wall transfer protein 1). A novel echinocandin called rezafungin is also being tested in these animal models as a potential therapeutic to combat C. auris.[25–28] Because of its multidrug resistance, environmental persistence, and the large inoculum used when working with animal models infected with C. auris, the risk of an occupational exposure during experiments for researchers and animal care staff is still unclear.
Impacts of the prostate stem cell antigen (PSCA) and Clostridium perfringens enterotoxin (CPE) on the apoptosis and cell cycle regulatory genes in PC3
Published in Preparative Biochemistry & Biotechnology, 2020
Saied Abedi, Abbas Doosti, Mohammad-Saied Jami
The prostate stem cell antigen (PSCA) is a glycosylphosphatidylinositol-anchored 123 amino acid protein related to the Tye-1/Ly-6 family of cell surface proteins.[15] PSCA mRNA has been detected in more than 80% of primary prostate cancers and correlate with increasing tumor grade, stage, and especially has been seen in patients with bone metastasis.[16] PSCA is expressed by a gene located on chromosome eight inhuman. It has been shown that vaccination with PSCA, induces a protective immune response against prostate cancer.[17] It has been suggested that PSCA is involved in the cell-proliferation inhibition of prostate epithelial cell lines.[18–20]
Production of polyclonal antibody against human Neuritin and its application of immunodetection
Published in Preparative Biochemistry and Biotechnology, 2019
Na Wang, Yu Wei, Wen Zhang, Xingyi Li, Jingling Zhu, Liya Shan, Chunyan Liu, Wumei Yuan, Jin Huang
The neuritin gene is located in the 6p24-p25 interval on chromosome 6. It has an open reading frame of 426 bp and encodes a mature Neuritin of 142 amino acids. Residues 1–27 encode a signal peptide, 116–142 encode a glycosylphosphatidylinositol anchor sequence, and 28–115 encode an active fragment. Using Pichia pastoris, His-Neuritin was constructed,[11] expressed, and successfully identified. Bioinformatic software[12] was used to analyze B-cell epitopes of the Neuritin active fragment. The analysis using the ABCpred online server and BepiPred 1.0 server identified the same epitopes in the active fragment, which indicated that Neuritin could serve as an excellent immunogen.