Article
Chemistry, Multidisciplinary
Martin D. Gelenter, Ad Bax
Summary: Post-translational modifications are commonly found in eukaryotic proteomes. However, these modifications are rarely studied using NMR in eukaryotic proteins, which are usually produced in E. coli through recombinant expression. The native C-terminal amide of melittin, a major peptide in honey bee venom, significantly affects its structure and dynamics. This study presents a method to produce isotopically labeled native melittin through recombinant expression and chemical amidation. It also confirms the accuracy of structural models generated by AlphaFold-Multimer using experimental residual dipolar couplings.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2023)
Article
Agriculture, Multidisciplinary
Haiyang Zhang, Xuehan Li, Qi Liu, Jianan Sun, Francesco Secundo, Xiangzhao Mao
Summary: By constructing a PLD expression and secretion system in Bacillus subtilis and introducing a nonclassical secretory pathway, high-level expression strains can be rapidly screened and target proteins can be secreted. Under optimal fermentation conditions, the enzyme activity in the culture medium reached 0.35 U/mL, which was 2.05-fold higher than that of the Sec secretion pathway strains. Additionally, the use of cyclopentyl methyl ether as the organic phase in biphasic reaction media achieved a final conversion rate of 96.9%.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2021)
Article
Microbiology
Irene Beriotto, Christopher Icke, Yanina R. R. Sevastsyanovich, Amanda E. E. Rossiter, Giacomo Romagnoli, Silvana Savino, Freya J. J. Hodges, Jeffrey A. A. Cole, Allan Saul, Calman A. A. MacLennan, Adam F. F. Cunningham, Francesca Micoli, Ian R. R. Henderson
Summary: The autotransporter protein secretion system has been successfully applied to target the secretion of difficult to produce recombinant proteins to the bacterial cell surface and extracellular milieu. In this study, the system was used to secrete an industrially relevant protein, IrmA, with comparable physical properties to the protein produced intracellularly. Scaling up production resulted in a high yield of IrmA, consistent with current industrial processes.
MICROBIOLOGY SPECTRUM
(2023)
Article
Biotechnology & Applied Microbiology
Nairiti J. Sinha, Christopher J. Kloxin, Jeffery G. Saven, Grethe V. Jensen, Zvi Kelman, Darrin J. Pochan
Summary: Computational design of fully artificial peptides is extensively researched for novel nanostructures and biomaterials construction. Recombinant expression is a convenient tool for synthesizing large quantities of artificial peptides efficiently.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Mousa Naderi, Roghayeh Ghaderi, Jafar Khezri, Aliasghar Karkhane, Bijan Bambai
Summary: Protein secretion into the periplasmic space requires interactions between the amino-terminal signal sequence and secretion machinery components. Modifying the amino-terminal sequence can improve secretion efficiency. The hydrophobicity level and secondary structure of the signal peptide have an impact on secretion efficiency. Introducing aromatic residues in the signal sequence also reduces secretion levels.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
(2022)
Article
Biotechnology & Applied Microbiology
Julia Kleetz, Georgios Vasilopoulos, Simon Czolkoss, Meriyem Aktas, Franz Narberhaus
Summary: Escherichia coli is a common model bacterium with a membrane composition that can be readily tailored to include non-native lipids such as phosphatidylcholine (PC) for biotechnology applications.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Article
Microbiology
Christopher J. Day, Alvin W. Lo, Lauren E. Hartley-Tassell, M. Pilar Argente, Jessica Poole, Nathan P. King, Joe Tiralongo, Michael P. Jennings, Mark A. Schembri
Summary: This study focused on dissecting the receptor specificity of different CU fimbriae in Escherichia coli using a glycan array analysis approach with whole-bacterial-cell surface plasmon resonance. The research revealed new information on fimbrial specificity and provided a rapid and scalable system to define novel adhesin-glycan interactions underlying bacterial colonization and disease. The findings contribute to a better understanding of bacterial adhesion mechanisms and offer a promising strategy to combat antibiotic resistance through blocking adhesion.
Article
Biochemistry & Molecular Biology
Xiaoying He, Yan Li, Yehui Tao, Xuelian Qi, Ruiqi Ma, Honghua Jia, Ming Yan, Kequan Chen, Ning Hao
Summary: Sucrose phosphorylase (SPase, EC2.4.1.7) is an important enzyme for producing glycosylated compounds. In this study, an attempt was made to extracellularly express a recombinant SPase from Thermobacillus sp. in E. coli, but the enzyme was unexpectedly produced outside the cells. Truncation experiments showed that both termini of the enzyme are crucial for protein folding and enzymatic activity.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Biochemistry & Molecular Biology
Lingyu Zhu, Zitao Guo, Ji Zhang, Yuliang Yang, Chunyu Liu, Liang Zhang, Zhenghua Gu, Youran Li, Zhongyang Ding, Guiyang Shi
Summary: In this study, human arresten and canstatin were successfully recombinantly expressed in Escherichia coli, and their anti-angiogenesis effects on human umbilical vein endothelial cells (HUVECs) were demonstrated. The results showed that arresten and canstatin could inhibit HUVECs proliferation, migration, and tube formation, as well as downregulate the expression of vascular endothelial growth factor (VEGF) and the phosphorylation levels of PI3K and Akt, suggesting the inhibition of the PI3K/Akt signaling pathway. These findings have important implications for the soluble recombinant expression of human arresten and canstatin, and for cancer therapy.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Hend Okasha, Sami Mohamed Nasr, Safia Samir
Summary: The study successfully expressed the rCec-B peptide using a pET-based expression system with a simple purification step, demonstrating its anti-cancer effect on Hepatocellular Carcinoma Cell line (HCC). The purified peptide showed a safe dose on normal WI-38 cells at 1.57 mg/ml, and an IC50 of 25 µg/ml on HepG2 cells.
CURRENT PHARMACEUTICAL BIOTECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Shuangying Chao, Yuhang Liu, Ning Ding, Yue Lin, Qian Wang, Junwen Tan, Wei Li, Yang Zheng, Xuejun Hu, Junming Li
Summary: In this study, the expression system of anti-GFP VHHs in E. coli was optimized, and it was found that signal peptides like OmpA and PelB can efficiently enhance the solubility of VHHs without affecting their bioactivity. This provides a novel strategy for the industrial production of soluble VHHs and the research of other VHHs.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Chemistry, Multidisciplinary
Jordan P. Hempfling, Emily R. Sekera, Amar Sarkar, Amanda B. Hummon, Dehua Pei
Summary: Efficient and convenient method for producing recombinant proteins with free N-terminal cysteines has been reported. Site-specific labeling of the resulting proteins at the N-terminus was achieved through a novel genetic fusion and sequential enzymatic processing.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2022)
Article
Multidisciplinary Sciences
Chae Won Kang, Hyun Gyu Lim, Jaehyuk Won, Sanghak Cha, Giyoung Shin, Jae-Seong Yang, Jaeyoung Sung, Gyoo Yeol Jung
Summary: In this study, the authors developed a synthetic adaptation strategy to improve the production of a microbial consortium. By introducing a genetic circuit in one of the strains, they were able to control the production of a specific compound and acclimate the consortium for higher production.
NATURE COMMUNICATIONS
(2022)
Article
Chemistry, Analytical
Samantha Nunez, Maria Barra, Daniel Garrido
Summary: In this study, a molecular quantification method of free fucose was designed and developed using fluorescent Escherichia coli. The molecular circuit showed specificity against other monosaccharides and exhibited a linear response in the range of 0-45 mM. The biosensor was successfully tested on different concentrations of free fucose and the supernatant of Bifidobacterium bifidum JCM 1254, indicating its applicability in detecting free fucose.
Review
Biochemical Research Methods
Kasper Hettinga, Etske Bijl
Summary: The consumption of animal proteins, particularly dairy proteins, is linked to sustainability and animal welfare issues. The synthesis of recombinant milk proteins is gaining interest, but the cost-effectiveness and environmental impact of the process are still uncertain.
CURRENT OPINION IN BIOTECHNOLOGY
(2022)