Article
Biotechnology & Applied Microbiology
Amruta A. Shendge, Jacinta S. D'Souza
Summary: Expressing affinity-tagged recombinant proteins at a high yield is crucial for various applications. Escherichia coli remains the most popular expression system due to its advantages in growth, genetic manipulation, and cost. However, some proteins tend to form inclusion bodies when over-expressed in E. coli, leading to purification and yield challenges. This study focuses on solubilizing hydrophobic peptides and proteins with amphipathic helices using a combinatorial approach of chemical and physical methods.
MICROBIAL CELL FACTORIES
(2022)
Article
Biotechnology & Applied Microbiology
Xi Xie, Pei Wu, Xiaochen Huang, WenFeng Bai, Bowen Li, Ning Shi
Summary: Developing a novel solubilizing fusion tag XXA has shown improved soluble expression of inclusion bodies and highlighted the potential of using antifreeze protein as a solubilizing fusion tag for the first time.
MICROBIAL CELL FACTORIES
(2022)
Article
Microbiology
Xuefeng Li, Baorong Zhang, Quan Hu, Changchao Chen, Jiahua Huang, Lu Liu, Shengbin Wang
Summary: Fusion tag-induced formation of inclusion bodies provides a powerful means to express unstructured or toxic proteins. Our study illustrated that the aggregation hot spots in a fusion tag played important roles in mediating its insoluble expression. Efficient production of inclusion bodies could also be implemented by refining its primary structure to form a more stable beta-sheet with higher hydrophobicity.
MICROBIOLOGY SPECTRUM
(2023)
Article
Multidisciplinary Sciences
Shohei Nosaki, Tohru Terada, Akira Nakamura, Kei Hirabayashi, Yuqun Xu, Thi Bao Chau Bui, Takeshi Nakano, Masaru Tanokura, Takuya Miyakawa
Summary: The maltose-binding protein (MBP) fusion tag is commonly used for crystallization of proteins, such as the plant-specific BZR transcription factor AtBIL1/BZR1 in Arabidopsis thaliana through MBP-mediated crystallization technique. This study demonstrates the potential of MBP crystallization chaperone and validates the structure of MBP-fused AtBIL1/BZR1-DNA using crystal packings analysis and molecular dynamics simulations after removing the MBP. The methodology combining MBP-mediated crystallization and MD simulation shows promising capabilities in understanding protein-DNA recognition.
SCIENTIFIC REPORTS
(2021)
Review
Medicine, Research & Experimental
Vasanti Suvarna, Bhunesh Bore, Chaitanya Bhawar, Rashmi Mallya
Summary: Bioactive phytochemicals from natural sources have diverse therapeutic activities, but their application is hindered by drawbacks such as poor solubility and bioavailability. However, complexation with cyclodextrins provides a solution to these issues. This article summarizes the complexation of various phytochemicals with cyclodextrins and the impact on their stability and dissolution rate, leading to enhanced bioactivity.
BIOMEDICINE & PHARMACOTHERAPY
(2022)
Article
Pharmacology & Pharmacy
Abdul Ahad, Yousef A. Bin Jardan, Mohd Zaheen Hassan, Mohammad Raish, Ajaz Ahmad, Abdullah M. Al-Mohizea, Fahad Al-Jenoobi
Summary: The aim of this study was to improve the solubility and dissolution rate of eprosartan mesylate by preparing an inclusion complex with beta-cyclodextrin using microwave technique. The inclusion complex showed better drug release profile compared to the physical mixture or pure drug, indicating its potential to improve the solubility and bioavailability of eprosartan. Molecular modeling study confirmed the good affinity of eprosartan to beta-cyclodextrin. This research provides valuable insights into enhancing the solubility of eprosartan.
Article
Biotechnology & Applied Microbiology
Yanjuan Liu, Mengru Zhang, Hang Yuan, Zhurong Zou
Summary: The study enhanced the activity of Escherichia coli DnaJ by using a hyper-acidified protein fusion strategy, showing improved solubility and activity of target proteins. The hyper-acidified DnaJ variants not only aid in correct folding of proteins into soluble and active forms, but also enhance heat tolerance in E. coli or yeast.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Toshinari Ezawa, Yukiko Inagaki, Kinami Kashiwaba, Namiko Matsumoto, Hajime Moteki, Isamu Murata, Yutaka Inoue, Mitsutoshi Kimura, Masahiko Ogihara, Ikuo Kanamoto
Summary: This study evaluated the solubility of piperine in biorelevant media and its effects on intestinal contractions when presented in inclusion complexes with cyclodextrins. The results showed that inclusion complexes significantly suppressed intestinal contractility, with the PP/alpha CD complex exhibiting the highest solubility and efficacy in suppressing intestinal contractions.
Article
Biochemistry & Molecular Biology
Svetlana Bakholdina, Anna M. Stenkova, Evgenia P. Bystritskaya, Evgeniy Sidorin, Natalya Yu Kim, Ekaterina S. Menchinskaya, Tatiana Yu Gorpenchenko, Dmitry L. Aminin, Nikita A. Shved, Tamara F. Solov'eva
Summary: The study showed that cultivation temperature influences the conformational quality of Yersinia pseudotuberculosis phospholipase A(1) (PldA) in inclusion bodies, with optimal fluorescence and expression observed at 18 degrees C. Solubilization of PldA-GFP from inclusion bodies in urea and SDS leads to denaturation, revealing structural heterogeneity. The use of GFP as a fusion partner improves conformational quality but negatively impacts enzymatic activity.
Article
Plant Sciences
Yajing Wang, Qian Chen, Xianfeng Huang, Xiaojing Yan
Summary: Acacetin-loaded microemulsion was prepared and evaluated for efficient pharmacokinetics and non-invasive transdermal delivery. The optimized formulation showed improved solubility and skin absorption efficiency of acacetin, providing a new option for drug delivery.
PHARMACEUTICAL BIOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Xiang-Yu Chen, Hui-Wen Yang, Shao-Ming Chi, Lu-Lu Yue, Qiong Ruan, Ze Lei, Hong-You Zhu, Yan Zhao
Summary: In this study, inclusion complexes between docetaxel and alkylenediamine-modified beta-cyclodextrins were prepared, which significantly increased the water solubility of docetaxel and exhibited better cytotoxicity against cancer cells compared to docetaxel alone.
Article
Food Science & Technology
Monize A. B. Baldassa, Raphael V. R. Dias, Leandro C. Oliveira, Eloi Feitosa
Summary: This study investigates the interaction between cornstarch and Pluronic F127 aqueous mixtures and reveals weak interactions between them, indicating a small solubility of cornstarch and no change in the micelle structure of F127 by moderate amounts of cornstarch.
FOOD RESEARCH INTERNATIONAL
(2022)
Article
Microbiology
Kerri Beth Boggs, Kearstin Edmonds, Nicolas Cifuentes-Munoz, Farah El Najjar, Conny Ossandon, McKenna Roe, Chao Wu, Carole L. Moncman, Trevor P. Creamer, Gaya K. Amarasinghe, Daisy W. Leung, Rebecca Ellis Dutch
Summary: Human metapneumovirus (HMPV) is a major cause of respiratory diseases in children, immunocompromised individuals, and the elderly. There is currently no effective treatment available. HMPV inclusion bodies (IBs) are dynamic structures required for viral replication and transcription. Recent research has suggested that HMPV IBs possess liquid organelle properties and that the viral P protein may play a role in their formation.
Article
Spectroscopy
Vimalasruthi Narayanan, Manawwer Alam, Naushad Ahmad, Suganya Bharathi Balakrishnan, Vigneshkumar Ganesan, Esakkimuthu Shanmugasundaram, Brindha Rajagopal, Stalin Thambusamy
Summary: In this study, solid inclusion complexes of CA/CDs were incorporated into PVA nanofibers using electrospinning technique to enhance solubility and antibacterial activity. The physical properties of CA/CDs were examined and the formation of ICs was confirmed. PVA/CA/CDs demonstrated promising performance in terms of antibacterial activity and solubility enhancement.
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
(2021)
Article
Biotechnology & Applied Microbiology
Irene Barguilla, Ugutz Unzueta, Jose Vicente Carratala, Olivia Cano-Garrido, Antonio Villaverde, Alba Hernandez, Neus Ferrer-Miralles
Summary: Bacterial inclusion bodies (IBs) are protein complexes that appear under stress conditions and recent research has shown their potential as biomaterials for cell growth promotion. In this study, the cytotoxicity, oxidative stress, and genotoxicity of different protein formats were analyzed in eukaryotic cells. The results demonstrate that natural IBs and soluble protein nanoparticles are non-toxic materials for eukaryotic cells.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biochemical Research Methods
Sreejith Raran-Kurussi, David S. Waugh
ANALYTICAL BIOCHEMISTRY
(2016)
Article
Biochemistry & Molecular Biology
Rajesh Gumpena, George T. Lountos, Sreejith Raran-Kurussi, Joseph E. Tropea, Scott Cherry, David S. Waugh
Article
Chemistry, Physical
Nghia Tuan Duong, Sreejith Raran-Kurussi, Yusuke Nishiyama, Vipin Agarwal
JOURNAL OF PHYSICAL CHEMISTRY LETTERS
(2018)
Article
Biochemical Research Methods
George T. Lountos, Sreejith Raran-Kurussi, Bryan M. Zhao, Beverly K. Dyas, Terrence R. Burke, Robert G. Ulrich, David S. Waugh
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
(2018)
Article
Biochemical Research Methods
Nghia Tuan Duong, Sreejith Raran-Kurussi, Yusuke Nishiyama, Vipin Agarwal
JOURNAL OF MAGNETIC RESONANCE
(2020)
Article
Chemistry, Physical
Lokeswara Rao Potnuru, Nghia Tuan Duong, Sahil Ahlawat, Sreejith Raran-Kurussi, Matthias Ernst, Yusuke Nishiyama, Vipin Agarwal
JOURNAL OF CHEMICAL PHYSICS
(2020)
Article
Biochemical Research Methods
Lokeswara Rao Potnuru, Nghia Tuan Duong, Budaraju Sasank, Sreejith Raran-Kurussi, Yusuke Nishiyama, Vipin Agarwal
Summary: Proton-detected solid-state NMR at fast Magic Angle Spinning (MAS) is commonly used for structural characterization of molecules using dipolar couplings. Selective proton recoupling techniques have emerged for structural characterization through estimation of qualitative and quantitative distances. The study demonstrates that well-characterized CNnv sequences can be tailored for selective recoupling of proton spins. The C6(4)(1) recoupling sequence with a modified POST-element shows selective polarization transfer efficiencies between various proton spin pairs.
JOURNAL OF MAGNETIC RESONANCE
(2021)
Article
Biochemical Research Methods
Sreejith Raran-Kurussi, Sarawata B. Sharwanlal, Deepa Balasubramanian, Kaustubh R. Mote
Summary: The fusion tag NT* shows high expression and solubility when used as an N-terminal fusion partner for aggregation-prone peptides, but it is not as effective as the well-known fusion tag MBP in enhancing passenger protein solubility. Although NT* has higher overall expression levels, MBP remains superior in improving solubility. After purification, the effective yield of both MBP-fusion and NT*-fusion from the soluble fraction is comparable.
PROTEIN EXPRESSION AND PURIFICATION
(2022)
Article
Chemistry, Physical
Pravin P. Taware, Sreejith Raran-Kurussi, Kaustubh R. Mote
Summary: The CURD strategy allows for time savings in acquiring chemical-shift assignments and distance restraints for proteins under slow-moderate magic-angle spinning conditions. By using the recycle delay of one experiment to establish correlations for another experiment, this strategy does not require additional radio-frequency deposition or lengthy optimization procedures.
JOURNAL OF PHYSICAL CHEMISTRY B
(2022)
Article
Biochemical Research Methods
Sahil Ahlawat, Kaustubh R. Mote, Sreejith Raran-Kurussi, Vipin Agarwal
Summary: Band Selective Spectral Spin-Diffusion (BASS-SD) is a method that enables selective proton-proton contacts in proteins. By using low-amplitude proton spinlock radio frequency pulses and fast MAS frequency, BASS-SD allows for selective polarization exchange in fully protonated molecules. This method can be used to observe protein structures and provide useful constraints for protein structure determination.
JOURNAL OF MAGNETIC RESONANCE
(2022)
Article
Biochemistry & Molecular Biology
Sahil Ahlawat, Subbarao Mohana Venkata Mopidevi, Pravin P. Taware, Sreejith Raran-Kurussi, Kaustubh R. Mote, Vipin Agarwal
Summary: A new method for assigning aromatic spins in a fully protonated protein is reported. This method connects the aromatic side-chain to the backbone atoms using a low-power TOBSY sequence. The unambiguous assignment of aromatic spins allows for the characterization of 1H-1H distance restraints involving aromatic spins. This method has been demonstrated on a fully protonated U-[13C,15N] labeled GB1 sample at 95-100 kHz MAS.
JOURNAL OF STRUCTURAL BIOLOGY-X
(2023)
Article
Chemistry, Physical
Pravin P. Taware, Mukul G. Jain, Sreejith Raran-Kurussi, Vipin Agarwal, P. K. Madhu, Kaustubh R. Mote
Summary: Proteins rely on conformational changes to carry out function, and measuring these changes can provide insight into their function. In this study, the researchers combined different strategies to measure dipole-dipole couplings in nondeuterated systems at a high MAS frequency of 100 kHz. These strategies allow for the access of dipolar order parameters in a variety of systems at increasingly fast MAS frequencies.
JOURNAL OF PHYSICAL CHEMISTRY LETTERS
(2023)
Article
Biotechnology & Applied Microbiology
S. Sarkar, D. Balasubramanian, N. Velugonda, N. Ede, M. Srinivas, M. R. Katika, S. Raran-Kurussi, M. Jaiswal
Summary: A cost-effective and robust diagnosis of an infectious agent is crucial during a pandemic outbreak. This study developed an improved RT-LAMP assay for detecting SARS-CoV-2, which showed high consistency with RT-qPCR. The colorimetric LAMP method can be used for high-throughput screening and is beneficial for disease control in resource-limited settings.
APPLIED BIOCHEMISTRY AND MICROBIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Pragyan P. Parida, Deepa Saraswathi, Subbarao M. V. Mopidevi, Sreejith Raran-Kurussi
Summary: Tobacco etch virus Protease (TEVp) is a cysteine protease known for its specific proteolysis. However, its insolubility limits its application. This study successfully improved the solubility of TEVp by using a fusion construct and obtained a large amount of soluble TEVp. This research is important for further understanding the characteristics and applications of TEVp.
CURRENT RESEARCH IN STRUCTURAL BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Anamika Biswas, Sreejith Raran-Kurussi, Akash Narayan, Abhisek Kar, Purna Chandra Mashurabad, Mrinal Kanti Bhattacharyya, Kalyaneswar Mandal
Summary: AMA1 is a surface protein of Plasmodium sp. essential for red blood cell invasion, making it a potential vaccine candidate. Challenges in achieving correct folding led to an improved refolding method and validation of its functional activity.
BIOCHEMISTRY AND BIOPHYSICS REPORTS
(2021)