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Lactic acid bacteria heterologous expression

Adelene Ai-Lian Song, Lionel L A In, Swee Hua Erin Lim, Raha Abdul Rahim
Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors...
April 4, 2017: Microbial Cell Factories
Pamela Mancha-Agresti, Mariana Martins Drumond, Fillipe Luiz Rosa do Carmo, Monica Morais Santos, Janete Soares Coelho Dos Santos, Franco Venanzi, Jean-Marc Chatel, Sophie Yvette Leclercq, Vasco Azevedo
Lactococcus lactis is well documented as a promising candidate for development of novel oral live vaccines. It has been broadly engineered for heterologous expression, as well as for plasmid expression vector delivery, directly inside eukaryotic cells, for DNA vaccine, or as therapeutic vehicle. This work describes the characteristics of a new plasmid, pExu (extra chromosomal unit), for DNA delivery using L. lactis and evaluates its functionality both by in vitro and in vivo assays. This plasmid exhibits the following features: (1) a theta origin of replication and (2) an expression cassette containing a multiple cloning site and a eukaryotic promoter, the cytomegalovirus (pCMV)...
March 17, 2017: Molecular Therapy. Methods & Clinical Development
Agnieszka K Szczepankowska, Katarzyna Szatraj, Przemysław Sałański, Agnieszka Rózga, Roman K Górecki, Jacek K Bardowski
Lactic acid bacteria (LAB) are Gram-positive, nonpathogenic microorganisms that are gaining much interest as antigen producers for development of live vaccine vectors. Heterologous proteins of different origin have been successfully expressed in various LAB species, including Lactococcus lactis. Recombinant L. lactis strains have been shown to induce specific local and systemic immune responses against various antigens. Our study aimed at constructing a L. lactis strain expressing haemagglutinin of a Polish avian H5H1 influenza isolate and examining its effect on animals...
2017: BioMed Research International
S Langa, J L Arqués, M Medina, J M Landete
AIMS: The aim of this study was the coproduction in a single strain of the Gram-negative bacteriocin colicin V with other bacteriocins from lactic acid bacteria (LAB). METHODS AND RESULTS: Colicin V was expressed in Lactococcus and Enterococcus strains by replacing the colicin V leader peptide by the leader peptide and promoter of d-alanyl-d-alanine carboxypeptidase from Lactobacillus reuteri CECT925 in pNZ8048 (pNZ:LR-colV). The antimicrobial activity of colicin V against the indicator organism Escherichia coli DH5α in transformed strains was checked by agar diffusion assay and SDS-PAGE analysis...
March 6, 2017: Journal of Applied Microbiology
Zhi-Qiang Xiong, Qiao-Hui Wang, Ling-Hui Kong, Xin Song, Guang-Qiang Wang, Yong-Jun Xia, Hui Zhang, Yong Sun, Lian-Zhong Ai
Bile salt hydrolase (BSH) plays an essential role in the cholesterol-removing effect of lactic acid bacteria, which hydrolyze conjugated bile salts to amino acid and deconjugated bile salts. However, Lactobacillus casei lacks the bsh gene, which may make it highly sensitive to bile salt stress. We wanted to improve the BSH activity of L. casei for various food-industry applications (e.g., milk fermentation). Plate assay testing indicated that Lactobacillus plantarum AR113 has the highest BSH activity. We cloned and sequenced 4 bsh genes from the genome of L...
February 2017: Journal of Dairy Science
Caroline Fritsch, André Jänsch, Matthias A Ehrmann, Simone Toelstede, Rudi F Vogel
A high variety of plants that are used for food production contain esterified hydroxycinnamic acids. As their free forms display several benefits, like an enhanced absorption in human intestinal tract, anti-oxidative and anti-carcinogenic effects, an improved protein solubility and reduced discoloration, the microbial ability to cleave the ester bond is highly desired. In order to examine potential fermentation strains for this purpose, six different lactic acid bacteria and one bifidobacterial strain were screened for their ability to degrade esterified hydroxycinnamic acids because these strains are commonly used for fermentation of plant-based foods...
February 2017: Current Microbiology
Sirintra Themsakul, Namfon Suebwongsa, Baltasar Mayo, Marutpong Panya, Viraphong Lulitanond
The ability to serve as a delivery vehicle for various interesting biomolecules makes lactic acid bacteria (LAB) very useful in several applications. In the medical field, recombinant LAB expressing pathogenic antigens at different cellular locations have been used to elicit both mucosal and systemic immune responses. Expression-secretion vectors (ESVs) with a signal peptide (SP) are pivotal for protein expression and secretion. In this study, the genome sequence of Lactobacillus casei ATCC334 was explored for new SPs using bioinformatics tools...
October 2016: FEMS Microbiology Letters
Zhi Li, Xuan Liu, Yi Li, Xiqian Lan, Polly Hangmei Leung, Jiashen Li, Gang Li, Maobin Xie, Yanxia Han, Xiaofen Lin
Antimicrobial peptides, produced by innate immune system of hosts in response to invading pathogens, are capable of fighting against a spectrum of bacteria, viruses, fungi, parasites and cancer cells. Here, a recombinant silkworm AMP Bmattacin2 from heterologous expression is studied, indicating a broad spectrum of antibacterial activity and showing selective killing ability towards skin and colon cancer cells over their normal cell counterparts. For the purpose of biomedical application, the electrospinning fabrication technique is employed to load Bmattacin2 into PLLA nanofibrous membrane...
2016: Scientific Reports
Sarah O'Flaherty, Todd R Klaenhammer
Clostridium botulinum and Bacillus anthracis produce potent toxins that cause severe disease in humans. New and improved vaccines are needed for both of these pathogens. For mucosal vaccine delivery using lactic acid bacteria, chromosomal expression of antigens is preferred over plasmid-based expression systems, as chromosomal expression circumvents plasmid instability and the need for antibiotic pressure. In this study, we constructed three strains of Lactobacillus acidophilus NCFM expressing from the chromosome (i) the nontoxic host receptor-binding domain of the heavy chain of Clostridium botulinum serotype A neurotoxin (BoNT/A-Hc), (ii) the anthrax protective antigen (PA), and (iii) both the BoNT/A-Hc and the PA...
October 15, 2016: Applied and Environmental Microbiology
Han Jiang, Ping Li, Qing Gu
Bacteriocin, which is produced by lactic acid bacteria (LAB), has the potential to act as natural preservatives in the food industry. To develop strategies to overproduce such peptides, plantaricin NC8, a class IIb LAB bacteriocin that consists of two peptides, PLNC8α and PLNC8β, was successfully heterologously expressed in Escherichia coli BL21 (DE3). PLNC8α and PLNC8β peptides were expressed as His6-tag fusion proteins and were separated by Ni(2+) chelating affinity chromatography. To get the PLNC8α and PLNC8β peptides without extra amino acids in the N-terminus, the fusion proteins were cleaved by enterokinase and further purified using the Ni-NTA Sefinose™ Resin Kit...
November 2016: Protein Expression and Purification
Xiao Yan Chen, Clemens Levy, Michael G Gänzle
Exopolysaccharides from lactic acid bacteria may improve texture and shelf life of bread. The effect of exopolysaccharides on bread quality, however, depends on properties of the EPS and the EPS producing strain. This study investigated structure-function relationships of EPS in baking application. The dextransucrase DsrM and the reuteransucrase GtfA were cloned from Weissella cibaria 10M and Lactobacillus reuteri TMW1.656, respectively, and heterologously expressed in Escherichia coli. Site-directed mutagenesis of GtfA was generates reuterans with different glycosidic bonds...
December 19, 2016: International Journal of Food Microbiology
Thibault Allain, Nahla M Mansour, May M A Bahr, Rebeca Martin, Isabelle Florent, Philippe Langella, Luis G Bermúdez-Humarán
Food-grade lactic acid bacteria, such as lactobacilli, represent good candidates for the development of mucosal vectors. Indeed, they are generally recognized as safe microorganisms and some strains display beneficial effects (probiotics). In this study, we described a new lactobacilli in vivo expression (LIVE) system for the production and delivery of therapeutic molecules at mucosal surfaces. The versatility and functionality of this system was successfully validated in several lactobacilli species; furthermore, we assessed in vivo LIVE system in two different mouse models of human pathologies: (i) a model of therapy against intestinal inflammation (inflammatory bowel diseases) and (ii) a model of vaccination against dental caries...
July 2016: FEMS Microbiology Letters
Suttipong Sak-Ubol, Peenida Namvijitr, Phornsiri Pechsrichuang, Dietmar Haltrich, Thu-Ha Nguyen, Geir Mathiesen, Vincent G H Eijsink, Montarop Yamabhai
BACKGROUND: Heterologous production of hydrolytic enzymes is important for green and white biotechnology since these enzymes serve as efficient biocatalysts for the conversion of a wide variety of raw materials into value-added products. Lactic acid bacteria are interesting cell factories for the expression of hydrolytic enzymes as many of them are generally recognized as safe and require only a simple cultivation process. We are studying a potentially food-grade expression system for secretion of hydrolytic enzymes into the culture medium, since this enables easy harvesting and purification, while allowing direct use of the enzymes in food applications...
May 12, 2016: Microbial Cell Factories
Erica Pontonio, Jennifer Mahony, Raffaella Di Cagno, Mary O'Connell Motherway, Gabriele Andrea Lugli, Amy O'Callaghan, Maria De Angelis, Marco Ventura, Marco Gobbetti, Douwe van Sinderen
BACKGROUND: Among the oligosaccharides that may positively affect the gut microbiota, xylo-oligosaccharides (XOS) and arabinoxylan oligosaccharides (AXOS) possess promising functional properties. Ingestion of XOS has been reported to contribute to anti-oxidant, anti-bacterial, immune-modulatory and anti-diabetic activities. Because of the structural complexity and chemical heterogeneity, complete degradation of xylan-containing plant polymers requires the synergistic activity of several enzymes...
May 3, 2016: Microbial Cell Factories
Namfon Suebwongsa, Viraphong Lulitanond, Baltasar Mayo, Panjamaporn Yotpanya, Marutpong Panya
There is an increasing interest to develop various lactic acid bacteria (LAB) species as mucosal delivery vehicles, for which the development of a variety of cloning and expression systems for these bacteria is of primary importance. This study reports the complete nucleotide sequence of the cryptic plasmid pRCEID7.6 derived from the chicken probiotic LAB strain Lactobacillus casei TISTR1341. Sequence analysis and comparison showed that pRCEID7.6 is composed of nine putative open reading frames. The replicon origin of pRCEID7...
2016: SpringerPlus
Patrycja A Kobierecka, Barbara Olech, Monika Książek, Katarzyna Derlatka, Iwona Adamska, Paweł M Majewski, Elżbieta K Jagusztyn-Krynicka, Agnieszka K Wyszyńska
Campylobacter jejuni is the most frequent cause of human food-borne gastroenteritis and chicken meat is the main source of infection. Recent studies showed that broiler chicken immunization against Campylobacter should be the most efficient way to lower the number of human infections by this pathogen. Induction of the mucosal immune system after oral antigen administration should provide protective immunity to chickens. In this work we tested the usefulness of Lactococcus lactis, the most extensively studied lactic acid bacterium, as a delivery vector for Campylobacter antigens...
2016: Frontiers in Microbiology
Bing-cun Ma, Xin Yang, Hong-ning Wang, Hai-peng Cao, Peng-wei Xu, Meng-die Ding, Hui Liu
To obtain adhesive and safe lactic acid bacteria (LAB) strains for expressing heterologous antigens, we screened LAB inhabitants in intestine of Tibetan chickens by analyzing their adhesion and safety properties and the selected LAB was engineered to express heterologous antigen (UTEpi C-A) based on chromosomal integration strategy. We demonstrated that a new Lactobacillu salivarius TCMM17 strain is strongly adhesive to chicken intestinal epithelial cells, contains no endogenous plasmids, is susceptible to tested antimicrobials, and shows no toxicities...
2016: Bioscience, Biotechnology, and Biochemistry
Juan J Jiménez, Dzung B Diep, Juan Borrero, Loreto Gútiez, Sara Arbulu, Ingolf F Nes, Carmen Herranz, Luis M Cintas, Pablo E Hernández
BACKGROUND: Bacteriocins produced by lactic acid bacteria (LAB) attract considerable interest as natural and nontoxic food preservatives and as therapeutics whereas the bacteriocin-producing LAB are considered potential probiotics for food, human and veterinary applications, and in the animal production field. Within LAB the lactobacilli are increasingly used as starter cultures for food preservation and as probiotics. The lactobacilli are also natural inhabitants of the gastrointestinal (GI) tract and attractive vectors for delivery of therapeutic peptides and proteins, and for production of bioactive peptides...
2015: Microbial Cell Factories
Tannaz Jalilsood, Ali Baradaran, Adelene Ai-Lian Song, Hooi Ling Foo, Shuhaimi Mustafa, Wan Zuhainis Saad, Khatijah Yusoff, Raha Abdul Rahim
BACKGROUND: Bacterial biofilms are a preferred mode of growth for many types of microorganisms in their natural environments. The ability of pathogens to integrate within a biofilm is pivotal to their survival. The possibility of biofilm formation in Lactobacillus communities is also important in various industrial and medical settings. Lactobacilli can eliminate the colonization of different pathogenic microorganisms. Alternatively, new opportunities are now arising with the rapidly expanding potential of lactic acid bacteria biofilms as bio-control agents against food-borne pathogens...
2015: Microbial Cell Factories
Gargi Pal, Sheela Srivastava
Most bacteriocins of lactic acid bacteria (LAB) are initially synthesized as pre-peptides with an N-terminal extension (leader peptides). Generally, the precursor peptides containing a double-glycine-type leader are processed by a dedicated ATP-binding cassette (ABC) transporter. The ABC transporter and an accessory protein lead to the cleavage of inactive pre-peptide with the concomitant export of the mature peptide across the cytoplasmic membrane. Plantaricins E, F, J, and K belong to class IIb 2-peptide bacteriocins and are synthesized as pre-peptides containing N-terminal G-G leader peptide...
August 2015: Archives of Microbiology
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