Article
Agriculture, Multidisciplinary
Fen Yan, Junying Chen, Ting Cai, Jinfu Zhong, Shaolong Zhang
Summary: A novel bifunctional alginate lyase, BY17PV7, was cloned and expressed in Escherichia coli BL21(DE3). BY17PV7 showed the greatest activity at 43 degrees C and pH 8.9, with a higher affinity for polyG. It exhibited a wide range of substrate specificity and good degradation effects. Furthermore, the released AOS by BY17PV7 showed strong antioxidant ability.
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
(2022)
Article
Biochemistry & Molecular Biology
Jin Yang, Dandan Cui, Shuo Ma, Wenkang Chen, Diwen Chen, Hong Shen
Summary: AlgSH17, a novel alginate lyase from marine bacterium Microbulbifer sp. SH-1, demonstrates excellent cold-adapted property and both exolytic and endolytic activities. It can degrade alginate into monosaccharides and oligosaccharides, exhibiting high antioxidative and preservation effects, suggesting its potential applications in the production of monosaccharides, antioxidants, and food additives.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Chemistry, Medicinal
Rui Yin, Yan-Jun Yi, Zhuo Chen, Bao-Xun Wang, Xue-Han Li, Yan-Xia Zhou
Summary: In this study, a new exo-alginate lyase named Alys1 with high activity was cloned from a marine strain, showing cold-adapted characteristics and efficient degradation of alginates at low temperatures. Alys1 can depolymerize alginates polyM and polyG, producing monosaccharides as minimal alginate oligosaccharides, and demonstrates bifunctionality with a preference for M over G. These properties make Alys1 a valuable candidate for both basic research and industrial applications.
Article
Biochemistry & Molecular Biology
Hae-Rin Jeong, Ju-Soon Yoo, Yong-Lark Choi, Yu-Sin Jang, Yong-Suk Lee
Summary: Alginate and its derivatives are natural polymers that are annually produced in large quantities and applied to various industries. MtAl138, a newly discovered alginate lyase, shows outstanding enzymatic activity at concentrated salt solutions and organic solvents, making it a reasonably attractive enzyme for industrial use.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Chemistry, Medicinal
Mingpeng Wang, Lei Chen, Zhengyu Lou, Xueting Yuan, Guiping Pan, Xiaoyan Ren, Pengyu Wang
Summary: Alginate oligosaccharides (AOS) are low molecular weight alginate with improved water solubility and comprehensive health benefits. In this study, a novel alginate lyase called Algpt was discovered and characterized from a marine bacterium. Algpt exhibited high stability and adaptability, making it suitable for efficient preparation of AOS with different sizes and AOS-based products.
Article
Chemistry, Medicinal
Xiao-Meng Sun, Zhao Xue, Mei-Ling Sun, Yi Zhang, Yu-Zhong Zhang, Hui-Hui Fu, Yu-Qiang Zhang, Peng Wang
Summary: In this study, a novel alginate lyase, AlyC8, with two functional catalytic domains (CD1 and CD2) was discovered. Both CD1 and CD2 showed synergistic effects in alginate degradation. The results demonstrated that CD1 plays a major role in the degradation of alginate by AlyC8, and CD1 and CD2 have different substrate specificities.
Article
Biotechnology & Applied Microbiology
Liufei Long, Qingsong Hu, Xinxia Wang, Hebin Li, Zhipeng Li, Zedong Jiang, Hui Ni, Qingbiao Li, Yanbing Zhu
Summary: Alginate lyase AlgL6 from Microbulbifer sp. ALW1 is a new alginate lyase classified into PL6 family, with enzymatic activities towards various polysaccharides and good stability and halo-stability. The degradation products of sodium alginate by AlgL6 show enhanced antioxidant activities. Structural analysis reveals the catalytic mechanism of AlgL6 and the involvement of Ca2+ in its activity.
ENZYME AND MICROBIAL TECHNOLOGY
(2022)
Article
Chemistry, Medicinal
Li Sha, Minghai Huang, Xiaonan Huang, Yongtong Huang, Ensi Shao, Xiong Guan, Zhipeng Huang
Summary: A highly active novel alginate lyase was obtained from marine bacteria. The enzyme showed preference for polyM as substrate, and its activity was enhanced by K+ and inhibited by Fe2+ and Cu2+. It has promising industrial application prospects.
Article
Biochemical Research Methods
Jun Jiang, Zhengqiang Jiang, Qiaojuan Yan, Susu Han, Shaoqing Yang
Summary: A novel alginate lyase gene from a marine bacterium showed efficient enzyme activity and stability, producing high conversion ratios of alginate oligosaccharides.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemistry & Molecular Biology
Zhongbin Xiao, Kuikui Li, Tang Li, Fanxing Zhang, Jiayi Xue, Miao Zhao, Heng Yin
Summary: Alginate lyase VwAlg7A, a novel PL7 family exolytic alginate lyase, was successfully heterologously expressed in E. coli BL21 (DE3). VwAlg7A showed specificity towards poly-guluronate and exhibited optimal temperature and pH at 30 degrees C and 7.0, respectively. The activity of VwAlg7A could be inhibited by Ni2+, Zn2+, and NaCl, and its kinetic parameters were determined as K-m of 36.9 mg/ml and V-max of 395.6 mu M/min. ESI and HPAEC-PAD results confirmed the exolytic mode of sugar bond cleavage by VwAlg7A, and important catalytic residues were identified through molecular docking and mutagenesis experiments.
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Fengchao Zhang, Zheng Fu, Luyao Tang, Zhelun Zhang, Feng Han, Wengong Yu
Summary: The novel exo-type alginate lyase VsAly7D isolated from brown algae showed the highest activity at 35 degrees C, with considerable activity at 20 and 30 degrees C, while exhibiting optimal activity under alkaline conditions. VsAly7D demonstrated efficient degradation of alginate, polyM, and polyG, making it a promising tool enzyme for bioethanol production from brown algae.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biotechnology & Applied Microbiology
Zhipeng Li, Xiaoyi Huang, Yuxi Guo, Chenghao Zhang, Liang Yang, Xiping Du, Hui Ni, Xuchu Wang, Yanbing Zhu
Summary: In this study, a mass spectrometry-based quantitative analysis was used to study the degradation pathway of alginate polysaccharide in the bacterial strain Microbulbifer sp. ALW1. The results showed that ALW1 could effectively degrade alginate polysaccharide into disaccharides and trisaccharides within 12 hours. Proteome analysis identified numerous proteins involved in the digestion and metabolism of alginate polysaccharide. This study provides important insights into the genetic basis of alginate catabolism in ALW1 and may guide the utilization of this bacterial strain for efficient production of alginate oligosaccharides.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Microbiology
Zedong Jiang, Liufei Long, Meifang Liang, Hebin Li, Yanhong Chen, Mingjing Zheng, Hui Ni, Qingbiao Li, Yanbing Zhu
Summary: In this study, a new beta-glucosidase MaGlu1A was cloned from a marine bacterium, showing optimal activity at 40 degrees C and pH 4.5, substrate preference for aryl-beta-glycosidic bonds with glucose, fucose, and galactose moieties, and strong stimulation by supplemental glucose. The enzymatic characterization of MaGlu1A provides valuable information for its practical applications.
MICROBIOLOGICAL RESEARCH
(2021)
Article
Biochemical Research Methods
Hebin Li, Xiaoyi Huang, Shuxiang Yao, Chenghao Zhang, Xuan Hong, Ting Wu, Zedong Jiang, Hui Ni, Yanbing Zhu
Summary: AlgL7, an alginate lyase from Microbulbifer sp. ALW1, belongs to the polysaccharide lyase family 7 (PL7) and exhibits optimal activity at 40 degrees C and pH 7.0. It is stable at temperatures below 35 degrees C and within a pH range of 5.0-10.0, and shows resistance to certain reagents and surfactants. AlgL7 generates alginate oligosaccharides with antioxidant activities when degrading sodium alginate and can directly digest Laminaria japonica to produce alginate oligosaccharides. These findings suggest the potential application of AlgL7 in the high-value utilization of brown algae resources.
PROTEIN EXPRESSION AND PURIFICATION
(2022)
Article
Chemistry, Medicinal
Zhao Xue, Xiao-Meng Sun, Cui Chen, Xi-Ying Zhang, Xiu-Lan Chen, Yu-Zhong Zhang, Shou-Jin Fan, Fei Xu
Summary: Alginate oligosaccharides (AOS) have various biological activities and important applications, and alginate lyases have unique advantages in the preparation of AOS. In this study, a strain of Pseudoalteromonas arctica M9 was isolated from Sargassum, and five putative alginate lyases were identified in its genome. Among them, AlyM2 showed the highest activity and mainly produced trisaccharides from sodium alginate, indicating its potential for trisaccharide preparation.
Review
Biochemical Research Methods
Sara B. Linden, Adit B. Alreja, Daniel C. Nelson
Summary: The decline in new antibiotic candidates and the increase in antibiotic resistance have led to the development of alternative antimicrobials, such as bacteriophage-encoded endolysins. Current studies focus on the discovery, structure/function, engineering, immunogenicity, toxicity/safety, and efficacy of endolysins, with some already in human clinical trials. Streptococcal diseases primarily affect accessible mucous membranes, making them a promising target for protein therapeutics, such as targeted endolysin therapy.
CURRENT OPINION IN BIOTECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Sebastian S. Broendum, Daniel E. Williams, Brooke K. Hayes, Felix Kraus, James Fodor, Ben E. Clifton, Anne Geert Volbeda, Jeroen D. C. Codee, Blake T. Riley, Nyssa Drinkwater, Kylie A. Farrow, Kirill Tsyganov, Ryan D. Heselpoth, Daniel C. Nelson, Colin J. Jackson, Ashley M. Buckle, Sheena McGowan
Summary: The study identified the polyrhamnose backbone of Group A streptococcal cell wall as the binding target of PlyC, and characterized the putative rhamnose binding groove of PlyC with critical residues. The interaction between PlyC and the cell wall may be a high avidity one rather than a high-affinity one, contributing to PlyC's remarkable lytic activity.
MOLECULAR MICROBIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Harley King, Sowmya Ajay Castro, Amol Arunrao Pohane, Cynthia M. Scholte, Vincent A. Fischetti, Natalia Korotkova, Daniel C. Nelson, Helge C. Dorfmueller
Summary: Endolysins are phage proteins that interact with bacterial cell walls through enzymatically active domains and cell wall binding domains, with the ability to lyse Gram-positive bacterial cells. They offer a potential alternative to traditional antibiotics, but research on the nature of CBDs and binding epitopes is still limited.
BIOCHEMICAL JOURNAL
(2021)
Article
Instruments & Instrumentation
Kyle W. Anderson, Kerry Scott, Ioannis L. Karageorgos, Elyssia S. Gallagher, Venkata S. Tayi, Michael Butler, Jeffrey W. Hudgens
JOURNAL OF RESEARCH OF THE NATIONAL INSTITUTE OF STANDARDS AND TECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Kyle W. Anderson, Christina Bergonzo, Kerry Scott, Ioannis L. Karageorgos, Elyssia S. Gallagher, Venkata S. Tayi, Michael Butler, Jeffrey W. Hudgens
Summary: Previous studies have proposed different models for stabilizing the Fc-Fc gamma RI immune complex. This study used multiple experimental methods to investigate the role of glycoprotein interactions between IgG1 glycans and the FG-loop of Fc gamma RIa in complex stabilization. The results showed that both intermolecular and intramolecular glycoprotein bonds contribute to the stability of the complex.
JOURNAL OF MOLECULAR BIOLOGY
(2022)
Article
Multidisciplinary Sciences
Kinlin L. Chao, Xiaoran Shang, Julia Greenfield, Sara B. Linden, Adit B. Alreja, Daniel C. Nelson, Osnat Herzberg
Summary: Four tailspike proteins enable infection of multiple bacterial hosts by the Escherichia coli O157:H7 bacteriophage CBA120. The crystal structure of one of these proteins, TSP4-N-335, reveals its trimeric nature and its role in anchoring the complex to the tail baseplate. Further analyses show that different regions of TSP4 are responsible for binding to other proteins. A 3-dimensional model of the bacteriophage CBA120 TSP complex has been developed based on these findings.
SCIENTIFIC REPORTS
(2022)
Article
Microbiology
Marek Harhala, Katarzyna Gembara, Paulina Miernikiewicz, Barbara Owczarek, Zuzanna Kazmierczak, Joanna Majewska, Daniel C. Nelson, Krystyna Dabrowska
Summary: Endolysins are considered promising solutions among emerging therapies for multi-drug resistant bacterial infections. However, there are still research issues to be clarified, including the indirect and inefficient methods for testing endolysin kinetics. Researchers have demonstrated a novel application of a DNA dye, SYTOX Green, for comprehensive, real-time, and rapid measurement of killing efficiency, lytic activity, and bacterial susceptibility to lytic enzymes.
FRONTIERS IN MICROBIOLOGY
(2021)
Article
Immunology
Meghna Thakur, Sandhya Bista, Shelby D. Foor, Shraboni Dutta, Xiuli Yang, Michael Ronzetti, Vipin S. Rana, Chrysoula Kitsou, Sara B. Linden, Amanda S. Altieri, Bolormaa Baljinnyam, Daniel C. Nelson, Anton Simeonov, Utpal Pal
Summary: The protein BB0323 of Borrelia burgdorferi undergoes a proteolytic maturation event mediated by the protease BbHtrA, which cleaves BB0323 between specific amino acids. Mutations at these cleavage sites affect the interaction between BB0323 and BbHtrA, leading to impaired infection ability of Borrelia burgdorferi.
INFECTION AND IMMUNITY
(2022)
Article
Microbiology
Ryan D. Heselpoth, Chad W. Euler, Vincent A. Fischetti
Summary: This study evaluates the antibacterial properties of a modified cationic peptide, PaP1, derived from the broad-acting lysin PlyPa01. PaP1 exhibits potent in vitro bactericidal activity against high priority Gram-positive and Gram-negative pathogens, including antibiotic-resistant strains. The peptide is effective against both planktonic and biofilm-state bacteria, and it is non-toxic to human cells. In addition, PaP1 enhances the potency of topical antibiotics in treating chronic wound infections in a murine model.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biochemical Research Methods
Kyle W. Anderson, Jeffrey W. Hudgens
Summary: To improve the reproducibility of HDX-MS in the quality control of biopharmaceuticals, researchers have developed a dual-enzyme HDX-MS instrument that conducts liquid chromatography separations at subzero temperature. This instrument reduces back-exchange and supports longer separations with improved resolution.
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
(2022)
Article
Infectious Diseases
Marek Adam Harhala, Katarzyna Gembara, Daniel C. Nelson, Paulina Miernikiewicz, Krystyna Dabrowska
Summary: This study evaluated the immunogenicity of PlyC, an antibacterial enzyme, in humans and mice. PlyC induced efficient IgG production in mice and interacted specifically with the PlyCA subunit. Approximately 10% of the human population showed immune reactivity to the PlyCB subunit, but no hypersensitivity reactions were observed.
Article
Pharmacology & Pharmacy
Yuncheng Wang, Dipesh Khanal, Adit B. Alreja, Hang Yang, Rachel Y. K. Chang, Waiting Tai, Mengyu Li, Daniel C. Nelson, Warwick J. Britton, Hak-Kim Chan
Summary: This study investigated the feasibility of producing inhalable powders of endolysins Cpl-1 and ClyJ-3 using spray drying method with excipients. The results showed that ClyJ-3 lost its bioactivity before the drying process, while Cpl-1 maintained its bioactivity in the spray-dried powders. Cpl-1 formulations with leucine and either lactose or trehalose exhibited promising physico-chemical properties and aerosol performances. These findings suggest that endolysin Cpl-1 can be formulated as spray dry powders for potential treatment of pulmonary infections.
INTERNATIONAL JOURNAL OF PHARMACEUTICS
(2023)
Article
Chemistry, Medicinal
Adit B. Alreja, Sara B. Linden, Harrison R. Lee, Kinlin L. Chao, Osnat Herzberg, Daniel C. Nelson
Summary: The study reveals the molecular mechanisms of Cpl-1 and confirms the involvement of the C-terminal tail region in the formation of a dimer. Key residues that mediate Cpl-1 dimer formation are identified. These findings shed light on the mechanisms of Cpl-1 and related enzymes and can guide future engineering efforts for therapeutic development against S. pneumoniae.
ACS INFECTIOUS DISEASES
(2023)
Article
Biochemical Research Methods
Kyle W. Anderson, Jeffrey W. Hudgens
Summary: Chromatographic separations at subzero temperature improve the precision of back-exchange-corrected HDX-MS determinations. A dual-enzyme HDX-MS analysis instrument with a RPLC trap column and a HILIC analytical column in a two-dimensional RPLC-HILIC configuration at subzero temperature was designed and performed. The HILIC column at -30°C showed reduced backpressure, enabling faster flow rates and the use of materials rated for lower maximum pressures.
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
(2023)