Article
Agriculture, Multidisciplinary
Bibek Byanju, Swastik Sen, Thomas Mansell, Buddhi P. P. Lamsal
Summary: Corn steep liquor (CSL) and thin stillage were evaluated as growth media for recombinant Lactococcus lactis to produce antifreeze proteins (AFPs). Light CSL and thin stillage were optimized with additives and trace elements to enhance the growth of wild-type and recombinant strains. The fermentation supernatants showed longer times to supercool and freeze, indicating potential use as antifreeze compounds in frozen food and non-food applications.
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
(2023)
Article
Biotechnology & Applied Microbiology
Yaqian Ai, Xingjiang Li, Xuefeng Wu, Manuel Montalban-Lopez, Zhi Zheng, Dongdong Mu
Summary: Barnase is an enzyme used for plasmid purification, targeted gene therapy, and protein interaction studies. The barnase gene from Bacillus amyloliquefaciens BH072 was successfully cloned into Lactococcus lactis, and its secretion was controlled using signal peptides. The optimal conditions for barnase activity and enzymatic reduction of nucleic acids in SCP forages were investigated. Homology modeling of barnase revealed its three-dimensional structure and substrate simulation docking predicted key active residues and bonding patterns.
ENZYME AND MICROBIAL TECHNOLOGY
(2023)
Article
Microbiology
Gabriel A. Monteiro, Sofia O. D. Duarte
Summary: This article reviews the major findings from transcriptome and proteome studies of Lactococcus lactis and explores how to improve the yield and quality of plasmid DNA or recombinant proteins through genetic engineering. The study found that lactococcus lactis harboring high copy numbers of plasmids exhibited different proteome profiles compared to strains with low copy numbers. It was also found that cell-wall anchored antigens showed more promising results in mucosal vaccination applications. However, engineering lactococcus lactis to express membrane proteins increased cellular burden. Genome engineering strategies were suggested to enhance the profitability of the process. The article also includes studies on the effect of protein overexpression on transcriptome and proteome in Escherichia coli and Bacillus subtillis.
Article
Biotechnology & Applied Microbiology
Ayse Biyikli, Ramazan Nicin, Enes Dertli, Omer Simsek
Summary: In this study, the 4,6-alpha-GTase and 4,3-alpha-GTase genes from Limosilactobacillus reuteri E81 and Limosilactobacillus fermentum PFC282, respectively, were successfully expressed extracellularly using Lactococcus lactis as a host. Western blot screening confirmed the secretion of the enzymes, and NMR analysis demonstrated the formation of new bonds within the enzymes. These enzymes exhibited activity on starch and maltodextrins, and their production is important for starch retrogradation delay and glycaemic index reduction.
ENZYME AND MICROBIAL TECHNOLOGY
(2023)
Article
Virology
Kaige Zhai, Zhongwang Zhang, Xinsheng Liu, Jianliang Lv, Liping Zhang, Jiahao Li, Zhongyuan Ma, Yonglu Wang, Huichen Guo, Yongguang Zhang, Li Pan
Summary: This study utilized Lactococcus lactis as a genetically engineered vaccine and successfully constructed two recombinant strains capable of expressing PDCoV-S1 and PDCoV-S1-Co1 proteins. The immunogenic capacity of these strains was evaluated in mice, showing the potential of Lactococcus as a vaccine option for PDCoV.
Article
Biotechnology & Applied Microbiology
Nathalie N. S. E. Henriksen, Mads Frederik Hansen, Heiko T. Kiesewalter, Jakob Russel, Joseph Nesme, Kevin R. Foster, Birte Svensson, Gunnar Oregaard, Jakob Herschend, Mette Burmolle
Summary: This study shows that cultivation in a structured environment facilitates coexistence, evolution, and adaptation of bacterial communities. Co-cultivation in a biofilm model resulted in stable coexistence of two bacterial species and drove the evolution of high-yield variants. This study emphasizes the importance of structured environments for the cultivation and evolution of industrially important bacterial communities.
NPJ BIOFILMS AND MICROBIOMES
(2022)
Article
Immunology
Xiaoting Zhang, Ruihua Zhang, Jingyu Wang, Nana Sui, Guige Xu, Hui Yan, Yanli Zhu, Zhijing Xie, Shijin Jiang
Summary: The study developed a secretory recombinant L. lactis (MG1363-VP1) expressing the VP1-eGFP fusion protein of DHAV-1, which significantly induced specific immune responses in ducklings. Oral administration of the recombinant L. lactis stimulated both humoral and mucosal immune systems, resulting in protection against DHAV-1 infection and a reduction in mortality rates. The recombinant L. lactis provides a new approach for preventing and controlling DHAV-1 infection in duck farming.
Article
Biotechnology & Applied Microbiology
Hoda Shirdast, Fatemeh Ebrahimzadeh, Amir Hossein Taromchi, Yousef Mortazavi, Abdolreza Esmaeilzadeh, Mohammad Hadi Sekhavati, Keivan Nedaei, Esmat Mirabzadeh
Summary: Immunization using recombinant L. lactis expressing Omp31 induces both humoral and cellular immune responses in mice, showing the promising potential as an intervention against Brucella melitensis infections.
PROBIOTICS AND ANTIMICROBIAL PROTEINS
(2021)
Article
Biotechnology & Applied Microbiology
Pandeeswari Jeeva, Sruthi Rayadurgam Jayaprakash, Guhan Jayaraman
Summary: This study investigates the effect of hemin-supplemented respiratory metabolism on the production of high molecular weight hyaluronic acid (HA) in Lactococcus lactis. The results show that increasing dissolved oxygen levels lead to a steady increase in HA titer. Cofactor availability is strongly correlated with intracellular HA precursor concentrations, and hemin-supplemented cultures have the highest HA titer. The molecular weight of HA is strongly correlated with the ratio of precursor concentrations and inversely correlated with HA titer.
BIOCHEMICAL ENGINEERING JOURNAL
(2022)
Article
Immunology
Nan Wang, Jiahao Li, Yajun Wang, Yingying Wang, Defeng Zhang, Cunbin Shi, Yingying Li, Sven M. Bergmann, Xubing Mo, Jiyuan Yin, Qing Wang
Summary: In this study, Lactococcus lactis was used as an oral vaccine delivery system to express the VP6 protein of grass carp reovirus II (GCRV II). It was found that the recombinant L. lactis strain could induce mucosal immune protection, enhance nonspecific immune responses, and elicit both systemic and mucosal immunity in fish. This study suggests that L. lactis is a promising candidate for oral vaccine delivery in the aquaculture industry.
FRONTIERS IN IMMUNOLOGY
(2022)
Article
Food Science & Technology
Jiahong Li, Shiwen Yu, Xiaohua Pan, Ming Zhang, Zhuwu Lv, Li-Long Pan, Jia Sun
Summary: Recombinant CRAMP-producing L. lactis NZ9000 was developed and shown to protect against colitis, restore gut barrier, regulate cytokine levels, and exert therapeutic effects by inhibiting p38/NF-kappa B signaling pathway.
FOOD & NUTRITION RESEARCH
(2021)
Article
Virology
Ruihua Zhang, Tong Xu, Ziping Li, Longfei Li, Chunhong Li, Xinrui Li, Zhiyue Wang, Shaohua Wang, Xuejing Wang, Hongliang Zhang
Summary: In this study, a novel Lactococcus lactis (L. lactis) strain was constructed to express a recombinant fusion protein consisting of the HA1 proteins derived from an endemic H9N2 virus strain and chicken IgY Fc fragment. The results showed that chickens immunized with L. lactis HA1-Fc strain exhibited significantly enhanced immune responses and reduced viral titers, indicating its potential as an improved vaccine for controlling H9N2 avian influenza outbreaks.
Article
Chemistry, Applied
Min Li, Weicheng Li, Dongyu Li, Juanjuan Tian, Luyao Xiao, Lai-Yu Kwok, Wei Li, Zhihong Sun
Summary: Exopolysaccharides (EPSs) from Lactococcus lactis subsp. lactis IMAU11823 were found to have strong reducing power and no cytotoxicity. The EPSs had different sugar compositions and molecular weights, with EPS-1 being a linear structure composed of glucose and mannose, and EPS-2 being a more complex, non-linear structure composed of mannose, glucose, and rhamnose. Furthermore, a biosynthesis model for EPS production in the IMAU11823 strain was proposed. This study broadens the understanding of the formation, structure, and function of complex EPSs produced by Lactococcus lactis subsp. lactis IMAU11823.
Article
Food Science & Technology
Xin Shen, Weicheng Li, Hongyu Cai, Shuai Guo, Min Li, Yangshuo Liu, Zhihong Sun
Summary: This study analyzed the fermentation capacity and metabolic profiles of 17 L. lactis subsp. lactis strains and found significant metabolic differences between strains with different fermentation rates. Peptides, esters, and tributyrin were identified as biomarkers to distinguish between fast and slow fermentation groups.
FOOD RESEARCH INTERNATIONAL
(2022)
Article
Oncology
Katarzyna Kaczmarek, Jerzy Wieckiewicz, Kazimierz Weglarczyk, Maciej Siedlar, Jarek Baran
Summary: This study investigated the use of genetically modified Lactococcus lactis bacteria for local delivery of human soluble TRAIL in CRC treatment. The results showed selective elimination of CRC cells and enhanced anti-tumor activity with oral administration of Metformin.
Article
Pharmacology & Pharmacy
Elisa Rioja-Blanco, Alberto Gallardo, Irene Arroyo-Solera, Patricia Alamo, Isolda Casanova, Ugutz Unzueta, Naroa Serna, Laura Sanchez-Garcia, Miquel Quer, Antonio Villaverde, Esther Vazquez, Xavier Leon, Lorena Alba-Castellon, Ramon Mangues
Summary: This study developed a novel protein nanoparticle, T22-DITOX-H6, which selectively delivers the diphtheria toxin cytotoxic domain to CXCR4(+) HNSCC cells. The nanoparticles effectively blocked tumor metastasis and recurrence without causing off-target toxicity.
Article
Chemistry, Multidisciplinary
Hector Lopez-Laguna, Ariana Rueda, Carlos Martinez-Torro, Lucia Sanchez-Alba, Jose Vicente Carratala, Jan Atienza-Garriga, Eloi Parlade, Julieta M. Sanchez, Naroa Serna, Eric Volta-Duran, Neus Ferrer-Miralles, David Reverter, Ramon Mangues, Antonio Villaverde, Esther Vazquez, Ugutz Unzueta
Summary: We have developed a novel protein assembly method based on a histidine-templated cysteine-coupling concept, which enables the spontaneous and efficient self-assembly of tagged proteins into monodisperse nanoparticles. This method can be used for one-step IMAC-based protein purification and for the formation of higher-order microparticulate materials as nanoparticle-releasing protein-only depots. The green biofabrication concept proposed here offers wide applicability for the further development of clinically usable protein materials.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2023)
Article
Oncology
Esperanza Medina-Gutierrez, Annabel Garcia-Leon, Alberto Gallardo, Patricia Alamo, Lorena Alba-Castellon, Ugutz Unzueta, Antonio Villaverde, Esther Vazquez, Isolda Casanova, Ramon Mangues
Summary: The overexpression of CXCR4 receptor in tumor tissue of most endometrial cancer (EC) patients, especially at advanced stages, provides a potential target for novel therapies. The study evaluated the efficacy of two CXCR4-targeted nanotoxins, T22-DITOX-H6 and T22-PE24-H6, in EC mouse models. The nanotoxins selectively induced apoptosis in CXCR4(+) EC cells, inhibited tumor growth, and improved survival without off-target toxicity.
Review
Biotechnology & Applied Microbiology
Eric Volta-Duran, Eloi Parlade, Naroa Serna, Antonio Villaverde, Esther Vazquez, Ugutz Unzueta
Summary: Protein-based nanocarriers are versatile and biocompatible drug delivery systems that can recruit multiple functions in a single modular polypeptide. However, their efficacy is hindered by the degradation of the internalized material in endo-lysosomes. This review discusses the mechanisms to evade lysosomal degradation and highlights the potential impact of peptide domains on the functionality of protein nanocarriers.
BIOTECHNOLOGY ADVANCES
(2023)
Article
Biotechnology & Applied Microbiology
Jose Vicente Carratala, Jan Atienza-Garriga, Hector Lopez-Laguna, Esther Vazquez, Antonio Villaverde, Julieta M. Sanchez, Neus Ferrer-Miralles
Summary: To improve the purification yield and purity of difficult proteins, a novel method has been developed using bile salts to supplement crude cell extracts, resulting in a substantial improvement in protein capture by affinity chromatography.
MICROBIAL CELL FACTORIES
(2023)
Article
Pharmacology & Pharmacy
Jose Luis Corchero, Marianna T. P. Favaro, Merce Marquez-Martinez, Jara Lascorz, Carlos Martinez-Torro, Julieta M. Sanchez, Hector Lopez-Laguna, Luis Carlos de Souza Ferreira, Esther Vazquez, Neus Ferrer-Miralles, Antonio Villaverde, Eloi Parlade
Summary: By using protein engineering techniques, recombinant proteins can be assembled into functional materials such as nanoparticles or secretory microparticles for drug delivery purposes. The use of histidine-rich tags and divalent cations allows the construction of these materials from pure polypeptide samples. The selection of the protein biofabrication platform is crucial in determining the biophysical properties of the resulting products.
Article
Pharmacology & Pharmacy
Adria Lopez-Cano, Neus Ferrer-Miralles, Julieta Sanchez, Jose Vicente Carratala, Xavier Rodriguez Rodriguez, Imma Ratera, Judith Guasch, Oscar Q. Q. Pich, Paula Bierge, Cristina Garcia-de-la-Maria, Jose M. M. Miro, Elena Garcia-Fruitos, Anna Aris
Summary: Antibiotic resistance has significantly increased, calling for the development of new antimicrobial drugs. Host Defense Peptides (HDPs) have the potential to be used as antimicrobial agents and immune regulators. The present study aims to design recombinant multidomain proteins based on HDPs to create a new generation of tailored antimicrobials. Three new antimicrobials were designed and tested, with D5L37D5L37 showing the highest efficacy against various healthcare-associated pathogens. This study highlights the potential of using HDP combinations for the development of new antimicrobial drugs.
Article
Pharmacology & Pharmacy
Yaiza Nunez, Annabel Garcia-Leon, Aida Falgas, Naroa Serna, Laura Sanchez-Garcia, Ana Garrido, Jorge Sierra, Alberto Gallardo, Ugutz Unzueta, Esther Vazquez, Antonio Villaverde, Ramon Mangues, Isolda Casanova
Summary: Researchers developed a protein nanoparticle, T22-PE24-H6, which can selectively deliver the exotoxin A to CXCR4(+) leukemic cells, and showed potent antitumor activity without toxicity.
Article
Nanoscience & Nanotechnology
Patricia Alamo, Eloi Parlade, Marianna T. P. Favaro, Alberto Gallardo, Rosa Mendoza, Luis C. S. Ferreira, Nerea Roher, Ramon Mangues, Antonio Villaverde, Esther Vazquez
Summary: Secretory protein microparticles are bio-inspired materials that mimic secretory granules in the human endocrine system. They are self-contained protein delivery platforms that slowly release proteins. By using cationic Zn for assembly, these granules are expected to be nontoxic and suitable for clinical use. However, their biosafety and protein dosage threshold have not been confirmed.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Engineering, Biomedical
Eric Volta-Duran, Lorena Alba-Castellon, Naroa Serna, Isolda Casanova, Hector Lopez-Laguna, Alberto Gallardo, Alejandro Sanchez-Chardi, Antonio Villaverde, Ugutz Unzueta, Esther Vazquez, Ramon Mangues
Summary: Researchers have developed biofabricated protein nanoparticles for targeted drug delivery into cancer-associated fibroblasts. These nanoparticles can selectively penetrate PDGFR-fi+ stromal fibroblasts and accumulate in tumor tissues, leading to tumor destruction without side effects.
ACTA BIOMATERIALIA
(2023)
Article
Agriculture, Dairy & Animal Science
M. Tortades, E. Garcia-Fruitos, A. Aris, M. Terre
Summary: Colostrum, the first milk, is crucial for neonatal survival due to its high nutrient concentrations and bioactive substances. This study aimed to measure the concentrations of IGF-I, IgG, and LTF in colostrum and transition milk and assess their potential use in veterinary and nutraceutical applications. The results showed a decrease in the concentrations of these bioactive molecules from the first milking to the tenth, with greater concentrations observed in multiparous cows. Transition milk exhibited a 46% decrease in the analyzed bioactive molecules. Further research is needed to apply this knowledge in neonate farm management or pharmaceutical supplements.
Article
Medicine, Research & Experimental
Carlos Martinez-Torro, Lorena Alba-Castellon, Laura Imedio Carrasco-Diaz, Naroa Serna, Laura Imedio, Alberto Gallardo, Isolda Casanova, Ugutz Unzueta, Esther Vazquez, Ramon Mangues, Antonio Villaverde
Summary: Researchers have engineered two human proteins, Gasdermin D (GSDMD) and Mixed Lineage Kinase Domain-Like (MLKL), to self-assemble with an efficient ligand of the tumoral cell marker CXCR4 and additional peptide agents. These proteins formed stable nanoparticles of 46 and 54 nm in diameter, showing moderate cytotoxicity in cell culture. When administered in mouse models of colorectal cancer, the nanoscale forms of GSDMD and MLKL promoted lymphocyte infiltration and reduced tumor size without systemic toxicity.
BIOMEDICINE & PHARMACOTHERAPY
(2023)
Article
Biochemistry & Molecular Biology
Eloi Parlade, Julieta M. Sanchez, Hector Lopez-Laguna, Ugutz Unzueta, Antonio Villaverde, Esther Vazquez
Summary: Hexahistidine-tagged proteins can form self-containing protein depots at the microscale, which can be used as drug delivery systems. The nature of the protein and the cross-linking cation used impact the disintegration and secretory performance of the depots. Factors such as electrostatic charge and protein properties influence the release kinetics and the fraction of releasable protein. Understanding these factors can help in designing efficient and customizable drug delivery platforms.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Article
Materials Science, Multidisciplinary
Ariana Rueda, Julian I. Mendoza, Lorena Alba-Castellon, Eloi Parlade, Eric Volta-Duran, David Paez, Anna Avino, Ramon Eritja, Esther Vazquez, Antonio Villaverde, Ramon Mangues, Ugutz Unzueta
Summary: This study reports a simple and straightforward method for the one-step production and site-directed cysteine conjugation of disulfide-containing non-antibody thiolated carrier proteins. The method allows for the development of stable, homogeneous, and efficient nanomedicines.
SCIENCE CHINA-MATERIALS
(2023)
Article
Pharmacology & Pharmacy
Naroa Serna, Hector Lopez-Laguna, Patricia Aceituno, Mauricio Rojas-Pena, Eloi Parlade, Eric Volta-Duran, Carlos Martinez-Torro, Julieta M. Sanchez, Angela Di Somma, Jose Vicente Carratala, Andrea L. Livieri, Neus Ferrer-Miralles, Esther Vazquez, Ugutz Unzueta, Nerea Roher, Antonio Villaverde
Summary: Both nanostructure and multivalency enhance the biological activities of antimicrobial peptides (AMPs). In addition, slow-release drug formulations based on nanostructured and recombinant AMPs have the potential to fight against bacterial infections.