Engineering Of Lactic Acid Bacteria For Delivery Of Therapeutic Proteins And Peptides
T. V. Plavec, A. Berlec
Published 2019 · Biology, Medicine
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Lactic acid bacteria (LAB) have a long-term history of use in food industry and are becoming attractive for use in therapy on account of their safety, intrinsic beneficial health effects, and considerable biotechnological potential. The established systems for engineering are combined with novel approaches, such as CRISPR-Cas, to enable the use of LAB as vectors for delivery of various therapeutic molecules. The latter are either secreted or surface displayed and can be used for the treatment or prevention of numerous conditions: inflammatory bowel diseases, infections, autoimmune diseases, and even cancer. This review presents some recent data on engineering of LAB, with the emphasis on the most commonly used genera Lactococcus and Lactobacillus. Their use for the delivery of therapeutic proteins is discussed, while a special focus is given to the delivery of therapeutic peptides. Therapeutically relevant improvements of engineered LAB, such as containment systems, ability to visualize bacteria, or target specific host cells are also addressed. Future engineering of LAB for therapy will adopt the capabilities of synthetic biology, with first examples already emerging.
This paper references
Development of a Stress-Inducible Controlled Expression (SICE) system in Lactococcus lactis for the production and delivery of therapeutic molecules at mucosal surfaces.
B. Benbouziane (2013)
Lactococcus lactis Expressing either Staphylococcus aureus Fibronectin-Binding Protein A or Listeria monocytogenes Internalin A Can Efficiently Internalize and Deliver DNA in Human Epithelial Cells
S. Innocentin (2009)
Display of recombinant proteins at the surface of lactic acid bacteria: strategies and applications
C. Michon (2016)
Mucosal and Systemic Immune Responses Induced by Recombinant Lactobacillus spp. Expressing the Hemagglutinin of the Avian Influenza Virus H5N1
Z. Wang (2011)
Engineering robust and tunable spatial structures with synthetic gene circuits
Wentao Kong (2017)
Signal Peptide and Propeptide Optimization for Heterologous Protein Secretion in Lactococcus lactis
Y. Le Loir (2001)
New thermosensitive plasmid for gram-positive bacteria.
E. Maguin (1992)
Targeted expression of anthrax protective antigen by Lactobacillus gasseri as an anthrax vaccine.
M. Mohamadzadeh (2010)
Fluorescent protein vectors for promoter analysis in lactic acid bacteria and Escherichia coli
Tomás García-Cayuela (2012)
2017b) CRISPR-Cas9(D10A) nickase-assisted genome editing in Lactobacillus casei
X Song (2017)
Berlec A (2015a) Heterologous surface display
P Zadravec (2015)
Effect of a mouth rinse formulation with human trefoil factor 1-secreting Lactococcus lactis in experimental oral mucositis in hamsters.
P. Rottiers (2009)
Oral delivery of Lactococcus lactis that secretes bioactive heme oxygenase-1 alleviates development of acute colitis in mice
Suguru Shigemori (2015)
Lactococcus lactis-based vaccines: Current status and future perspectives
Mohammed Bahey-El-Din (2011)
Genetic Tools for the Enhancement of Probiotic Properties.
Laura Ortiz-Velez (2017)
Immunization with r
B Yagnik (2017)
Design of a protein-targeting system for lactic acid bacteria.
Y. Dieye (2001)
Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis
S. Shigemori (2016)
Probiotic strains detect and suppress cholera in mice
Ning Mao (2018)
Microbially produced glucagon-like
A Narbad (2016)
Implementation of the agmatine-controlled expression system for inducible gene expression in Lactococcus lactis
D. Linares (2015)
Construction and Immunological Evaluation of Dual Cell Surface Display of HIV-1 Gag and Salmonella enterica Serovar Typhimurium FliC in Lactobacillus acidophilus for Vaccine Delivery
A. Kajikawa (2012)
Mucosal vaccination with recombinant poxvirus vaccines protects ferrets against symptomatic CDV infection.
J. Welter (1999)
Tunable protein degradation in bacteria
D. E. Cameron (2014)
Reversal of Diabetes in NOD Mice by Clinical-Grade Proinsulin and IL-10–Secreting Lactococcus lactis in Combination With Low-Dose Anti-CD3 Depends on the Induction of Foxp3-Positive T Cells
T. Takiishi (2016)
in recent - onset NOD mice
S Robert (2015)
A xylose-inducible expression system for Lactococcus lactis.
A. Miyoshi (2004)
Versatile Cas9-Driven Subpopulation Selection Toolbox for Lactococcus lactis
Simon van der Els (2018)
Microbially produced glucagon-like
T Arora (2016)
Construction of a new shuttle vector for Zymomonas mobilis
N. Misawa (1986)
A Novel Lactococcal Vaccine Expressing a Peptide from the M2 Antigen of H5N2 Highly Pathogenic Avian Influenza A Virus Prolongs Survival of Vaccinated Chickens
Kaleb A. Reese (2013)
Lactococcus lactis anchoring avian infectious
Enhanced heterologous protein productivity by genome reduction in Lactococcus lactis NZ9000
D. Zhu (2017)
Oral Delivery of Glutamic Acid Decarboxylase (GAD)-65 and IL10 by Lactococcus lactis Reverses Diabetes in Recent-Onset NOD Mice
S. Robert (2014)
Heterologous surface display on lactic acid bacteria: non-GMO alternative?
Petra Zadravec (2015)
Heterologous Protein Secretion in Lactobacilli with Modified pSIP Vectors
Ingrid Lea Karlskås (2014)
Zirex: a Novel Zinc-Regulated Expression System for Lactococcus lactis
Dongdong Mu (2013)
Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus siamensis Form an “Operational Group B. amyloliquefaciens” within the B. subtilis Species Complex
B. Fan (2017)
Mucosal Vaccination with Recombinant Lactobacillus casei-Displayed CTA1-Conjugated Consensus Matrix Protein-2 (sM2) Induces Broad Protection against Divergent Influenza Subtypes in BALB/c Mice
Mohammed Y. E. Chowdhury (2014)
Lactic acid bacteria — promising vaccine vectors: possibilities, limitations, doubts
K. Szatraj (2017)
In Vivo and In Vitro Detection of Luminescent and Fluorescent Lactobacillus reuteri and Application of Red Fluorescent mCherry for Assessing Plasmid Persistence
Shokoufeh Karimi (2016)
New Thermosensitive Delivery Vector and Its Use To Enable Nisin-Controlled Gene Expression in Lactobacillus gasseri
T. Neu (2003)
High efficiency recombineering
RA Britton (2012)
Anti‐cancer effect of lactic acid bacteria expressing antioxidant enzymes or IL‐10 in a colorectal cancer mouse model
S. Del Carmen (2017)
Synthetic Biology Approaches to Engineer Probiotics and Members of the Human Microbiota for Biomedical Applications.
Josef R. Bober (2018)
Lactic Acid Bacteria for Delivery of Endogenous or Engineered Therapeutic Molecules
P. Bron (2018)
Expression of green fluorescent protein in Lactococcus lactis.
P. Fernández de Palencia (2000)
Oral administration of a catalase
V Azevedo (2008)
Development of Recombinant Lactococcus lactis Displaying Albumin-Binding Domain Variants against Shiga Toxin 1 B Subunit
Petra Zadravec (2016)
In vitro and in vivo characterization of DNAdelivery using recombinant Lactococcus lactis expressing a mutated form of L. monocytogenes
M de Azevedo (2012)
Recombineering with overlapping single-stranded DNA oligonucleotides: Testing a recombination intermediate
Daiguan Yu (2003)
Microbially produced glucagon-like peptide 1 improves glucose tolerance in mice
T. Arora (2016)
In vivo imaging of Lactococcus lactis, Lactobacillus plantarum and Escherichia coli expressing infrared fluorescent protein in mice
Aleš Berlec (2015)
Isolation of strong constitutive promoters from Lactococcus lactis subsp. lactis N8.
D. Zhu (2015)
Intragastric administration of a superoxide dismutase-producing recombinant Lactobacillus casei BL23 strain attenuates DSS colitis in mice.
Laurie Watterlot (2010)
Mucosal vaccine delivery of antigens tightly bound to an adjuvant particle made from food-grade bacteria.
M. V. van Roosmalen (2006)
Orally administered L. lactis secreting an anti-TNF Nanobody demonstrate efficacy in chronic colitis
K. Vandenbroucke (2010)
LACTOBACILLUS CASEI のラット膀胱発癌に対する影響
K. Tomita (1994)
Recombinant Lactococcus lactis expressing porcine insulin-like growth factor I ameliorates DSS-induced colitis in mice
Shujie Liu (2016)
In vitro and in vivo characterization of DNA delivery using recombinant Lactococcus lactis expressing a mutated form of L. monocytogenes Internalin A
M. D. de Azevedo (2012)
Isolation of a novel IS3 group insertion element and construction of an integration vector for Lactobacillus spp.
D. Walker (1994)
Heme Oxygenase-1 Ameliorates Dextran Sulfate Sodium-induced Acute Murine Colitis by Regulating Th17/Treg Cell Balance*
Liya Zhang (2014)
Lactobacilli-Expressed Single-Chain Variable Fragment (scFv) Specific for Intercellular Adhesion Molecule 1 (ICAM-1) Blocks Cell-Associated HIV-1 Transmission across a Cervical Epithelial Monolayer1
Caren J. Chancey (2006)
Relevance and application of sortase and sortase-dependent proteins in lactic acid bacteria
E. Call (2013)
Induction of ovalbumin-specific tolerance by oral administration of Lactococcus lactis secreting ovalbumin.
I. Huibregtse (2007)
Oral vaccination of mice against tetanus with recombinant Lactococcus lactis
K. Robinson (1997)
Engineering recombinant Lactococcus lactis as a delivery vehicle for BPC-157 peptide with antioxidant activities
Katja Škrlec (2018)
Effect of recombinant Lactococcus lactis producing myelin peptides on neuroimmunological changes in rats with experimental allergic encephalomyelitis.
Kaja Kasarełło (2016)
Immunization with r‐Lactococcus lactis expressing outer membrane protein A of Shigella dysenteriae type‐1: evaluation of oral and intranasal route of administration
B. Yagnik (2017)
Lactobacillus casei secreting alpha-MSH induces the therapeutic effect on DSS-induced acute colitis in Balb/c Mice.
Sun-Woo Yoon (2008)
Recombinant Lactococcus lactis
S Liu (2016)
Cell Wall Anchoring of the Campylobacter Antigens to Lactococcus lactis
Patrycja A. Kobierecka (2016)
receiving induction chemotherapy
Y Lin (2016)
S Del Carmen (2017)
The European LABDEL project and its relevance to the prevention and treatment of allergies
C. Daniel (2007)
Improvement of LysM-Mediated Surface Display of Designed Ankyrin Repeat Proteins (DARPins) in Recombinant and Nonrecombinant Strains of Lactococcus lactis and Lactobacillus Species
Petra Zadravec (2015)
LG Bermudez-Humaran (2017)
Translocation of a Thioether-Bridged Azurin Peptide Fragment via the Sec Pathway in Lactococcus lactis
A. Kuipers (2009)
Lipoteichoic acid mediates binding of a Lactobacillus S-layer protein
Eva Bönisch (2018)
10 years of the nisin-controlled gene expression system (NICE) in Lactococcus lactis
I. Mierau (2005)
A phase I trial with transgenic bacteria expressing interleukin-10 in Crohn's disease.
H. Braat (2006)
p19-Targeting ILP Protein Blockers of IL-23/Th-17 Pro-Inflammatory Axis Displayed on Engineered Bacteria of Food Origin
Katja Škrlec (2018)
Oral Delivery of Pentameric Glucagon-Like Peptide-1 by Recombinant Lactobacillus in Diabetic Rats
Yin Lin (2016)
Development of a versatile Cas9-driven subpopulation-selection toolbox in Lactococcus lactis
S van der Els (2018)
Gene expression systems for lactic acid bacteria.
W. Vos (1999)
Lactic acid bacteria: reviewing the potential of a promising delivery live vector for biomedical purposes
O. Cano-Garrido (2015)
Construction and Characterization of Nisin-Controlled Expression Vectors for Use in Lactobacillus reuteri
Chi-Ming Wu (2006)
Efficacy of a Lactococcus lactis ΔpyrG vaccine delivery platform expressing chromosomally integrated hly from Listeria monocytogenes
Mohammed Bahey-El-Din (2010)
Serine protease inhibitors protect better than IL-10 and TGF-β anti-inflammatory cytokines against mouse colitis when delivered by recombinant lactococci
L. Bermúdez-Humarán (2015)
Microbial delivery vehicles for allergens
A Zahirovic (2018)
Development of Lactococcus lactis encoding
JG Gutierrez (2017)
Heterologous Expression and Delivery of Biologically Active Exendin-4 by Lactobacillus paracasei L14
Zhu Zeng (2016)
CRISPR-Cas9D10A Nickase-Assisted Genome Editing in Lactobacillus casei
X. Song (2017)
Oral immunization with live Lactococcus lactis expressing rotavirus VP8 subunit induces specific immune response in mice.
B. Marelli (2011)
CRISPR–Cas9-assisted recombineering in Lactobacillus reuteri
Jee-Hwan Oh (2014)
Living therapeutics: Scientists genetically modify bacteria to deliver drugs
Amy Maxmen (2017)
Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
Ali Baradaran (2016)
in NOD mice by clinical - grade proinsulin and IL - 10 - secreting Lactococcus lactis in combination with low - dose anti - CD 3 depends on the induction of Foxp 3 - positive T cells
M Toscano (2017)
A review on Lactococcus lactis: from food to factory
A. A. Song (2017)
Targeted expression of anthrax
V Cohran (2010)
Construction and characterization of nisin - controlled expression vectors for use in Lactobacillus reuteri
B Yagnik (2016)
Evasin‐displaying lactic acid bacteria bind different chemokines and neutralize CXCL8 production in Caco‐2 cells
Katja Škrlec (2017)
Positive role of cell wall anchored proteinase PrtP in adhesion of lactococci
Olivier Habimana (2006)
Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed That Editing Outcomes Can Vary Across Strains and Between Methods.
R. Leenay (2019)
Food-Grade Bacteria Expressing Elafin Protect Against Inflammation and Restore Colon Homeostasis
J. Motta (2012)
Protective immunity against influenza H5N1 virus challenge in chickens by oral administration of recombinant Lactococcus lactis expressing neuraminidase
Han Lei (2015)
Molecular characterization of the pH-inducible and growth phase-dependent promoter P170 of Lactococcus lactis.
S. M. Madsen (1999)
Protective effects of cathelicidin‐encoding Lactococcus lactis in murine ulcerative colitis
C. C. Wong (2012)
Critical Role of Antimicrobial Peptide Cathelicidin for Controlling Helicobacter pylori Survival and Infection
L. Zhang (2016)
Construction of a new shuttle vector for DNA delivery into mammalian cells using non-invasive Lactococcus lactis.
B. Yagnik (2016)
pSEUDO, a Genetic Integration Standard for Lactococcus lactis
João Carlos Pizani Pinto (2011)
Lactococcus lactis Anchoring Avian Infectious Bronchitis Virus Multi-Epitope Peptide EpiC Induced Specific Immune Responses in Chickens
Hai-peng Cao (2013)
A food-grade fimbrial adhesin FaeG expression system in Lactococcus lactis and Lactobacillus casei.
W. Lu (2016)
Intranasal immunization with recombinant Lactococci carrying human papillomavirus E7 protein and mouse interleukin-12 DNA induces E7-specific antitumor effects in C57BL/6 mice
Y. Li (2014)
JH Oh (2014)
HSP65 serves as an immunogenic carrier for a diabetogenic peptide P277 inducing anti-inflammatory immune response in NOD mice by nasal administration.
Jin Liang (2010)
Oral Immunization with a Recombinant Lactococcus lactis–Expressing HIV-1 Antigen on Group A Streptococcus Pilus Induces Strong Mucosal Immunity in the Gut
Venkateswarlu Chamcha (2015)
Recombinant Lactococcus lactis NZ9000 secretes a bioactive kisspeptin that inhibits proliferation and migration of human colon carcinoma HT-29 cells
Bo Zhang (2016)
Antimicrobial peptides targeting Gram-negative pathogens, produced and delivered by lactic acid bacteria.
Katherine Volzing (2013)
Engineering Signal Peptides for Enhanced Protein Secretion from Lactococcus lactis
Daphne T W Ng (2012)
A food-grade fimbrial
WW Lu (2016)
Oral Administration of Recombinant Lactococcus lactis Expressing HSP65 and Tandemly Repeated P277 Reduces the Incidence of Type I Diabetes in Non-Obese Diabetic Mice
Yanjun Ma (2014)
Single plasmid systems for inducible dual protein expression and for CRISPR-Cas9/CRISPRi gene regulation in lactic acid bacterium Lactococcus lactis
A. Berlec (2018)
Oral vaccination against HPV E7 for treatment of cervical intraepithelial neoplasia grade 3 (CIN3) elicits E7-specific mucosal immunity in the cervix of CIN3 patients.
K. Kawana (2014)
Development of Lactococcus lactis encoding fluorescent proteins, GFP, mCherry and iRFP regulated by the nisin-controlled gene expression system
Elvis Martinez-Jaramillo (2017)
Optimization of signal peptide SP310 for heterologous protein production in Lactococcus lactis.
P. Ravn (2003)
Trimming of two major type 1 diabetes driving antigens, GAD65 and IA-2, allows for successful expression in Lactococcus lactis.
S. Robert (2015)
Modulation of allergic immune responses by mucosal application of recombinant lactic acid bacteria producing the major birch pollen allergen Bet v 1
C. Daniel (2006)
Expression of a hepatitis A virus antigen in Lactococcus lactis and Escherichia coli and evaluation of its immunogenicity
Aleš Berlec (2013)
2016b) Critical role
Modulation of allergic immune responses
A Mercenier (2006)
Cathelicidin‐encoding Lactococcus lactis promotes mucosal repair in murine experimental colitis
C. C. Wong (2017)
Recombinant Lactococcus lactis co-expressing OmpH of an M cell-targeting ligand and IBDV-VP2 protein provide immunological protection in chickens.
L. Liu (2018)
Gene expression systems for lactic acid bacteria.
W. C. De Vos (1999)
Oral administration of a catalase-producing Lactococcus lactis can prevent a chemically induced colon cancer in mice.
A. de Moreno de LeBlanc (2008)
Controlled expression of CluA in Lactococcus lactis and its role in conjugation.
R. Stentz (2004)
The new frontier of genome engineering with CRISPR-Cas9
J. Doudna (2014)
Principles of genetic circuit design
Jennifer A N Brophy (2014)
Berlec A (2015b) Improvement of LysM
P Zadravec (2015)
Characterization of the protective
Engineering BmpA as a carrier for surface display of IgG-binding domain on Lactococcus lactis.
P. Zadravec (2014)
T Takiishi (2017)
Microbial Delivery Vehicles for Allergens and Allergen-Derived Peptides in Immunotherapy of Allergic Diseases
Abida Zahirović (2018)
Immobilization of Lactococcus lactis to cellulosic material by cellulose‐binding domain of Cellvibrio japonicus
K. Kylä-Nikkilä (2010)
Berlec A (2015a) Heterologous surface display on lactic acid bacteria: non-GMO alternative? Bioengineered
P Zadravec (2015)
High efficiency recombineering in lactic acid bacteria
Jan-Peter van Pijkeren (2012)
Receptor Binding Domain of Escherichia coli F18 Fimbrial Adhesin FedF Can Be both Efficiently Secreted and Surface Displayed in a Functional Form in Lactococcus lactis
A. Lindholm (2004)
Oral delivery of IL-27 recombinant bacteria attenuates immune colitis in mice.
Miranda L. Hanson (2014)
Prophylactic effect of Lactobacillus oral vaccine expressing a Japanese cedar pollen allergen.
Kayo Ohkouchi (2012)
Bioluminescence Imaging Study of Spatial and Temporal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice
C. Daniel (2012)
Modified Lactic Acid Bacteria Detect and Inhibit Multiresistant Enterococci
Juan Borrero (2015)
AG013, a mouth rinse formulation of Lactococcus lactis secreting human Trefoil Factor 1, provides a safe and efficacious therapeutic tool for treating oral mucositis.
S. Caluwaerts (2010)
Exploiting the peptidoglycan-binding motif, LysM, for medical and industrial applications
Ganesh Ram R. Visweswaran (2014)
Recombinant interleukin 6 with M cell-targeting moiety produced in Lactococcus lactis IL1403 as a potent mucosal adjuvant for peroral immunization.
Hui-Shan Li (2015)
Genetic tools for the enhancement
R Britton (2017)
Biological containment of genetically modified Lactococcus lactis for intestinal delivery of human interleukin 10
L. Steidler (2003)
Lactococcus lactis: high-level expression of tetanus toxin fragment C and protection against lethal challenge.
J. Wells (1993)
New Expression System Tightly Controlled by Zinc Availability in Lactococcus lactis
D. Llull (2004)
Lactic Acid Bacteria with Concomitant IL-17, IL-23 and TNFα- Binding Ability for the Treatment of Inflammatory Bowel Disease.
Stasa Kosler (2017)
Ability of Lactobacillus kefiri LKF01 (DSM32079) to colonize the intestinal environment and modify the gut microbiota composition of healthy individuals.
M. Toscano (2017)
Anti-inflammatory properties of Lactobacillus gasseri expressing manganese superoxide dismutase using the interleukin 10-deficient mouse model of colitis.
I. Carroll (2007)
Anti-inflammatory properties of C-Peptide.
Jaime Haidet (2009)
LG Bermudez-Humaran (2017)
tool for treating oral mucositis
DE Cameron (2014)
Genetically modified bacteria enlisted in fight against disease
Sara Reardon (2018)
Dendritic cell targeting of Bacillus anthracis protective antigen expressed by Lactobacillus acidophilus protects mice from lethal challenge
M. Mohamadzadeh (2009)
Generation of Lactococcus lactis capable of coexpressing epidermal growth factor and trefoil factor to enhance in vitro wound healing
Evanna Huynh (2015)
Phase 1b, multicenter, single blinded, placebo‐controlled, sequential dose escalation study to assess the safety and tolerability of topically applied AG013 in subjects with locally advanced head and neck cancer receiving induction chemotherapy
S. Limaye (2013)
Shigella IpaB and IpaD displayed on L. lactis bacterium-like particles induce protective immunity in adult and infant mice
Shannon J. Heine (2015)
Applications of Genetically Modified Immunobiotics with High Immunoregulatory Capacity for Treatment of Inflammatory Bowel Diseases
S. Shigemori (2017)
High-level, inducible gene expression in Lactobacillus sakei and Lactobacillus plantarum using versatile expression vectors.
E. Sørvig (2005)
Programmable probiotics for detection of cancer in urine
Tal Danino (2015)
Characterization of the protective immune response to Yersinia pseudotuberculosis infection in mice vaccinated with an LcrV-secreting strain of Lactococcus lactis.
Catherine Daniel (2016)
Lactic acid bacteria as mucosal delivery vehicles: a realistic therapeutic option
M. Wang (2016)
Lactobacilli-expressed single-chain variable
RB Markham (2006)
Bacterium-Like Particles for Efficient Immune Stimulation of Existing Vaccines and New Subunit Vaccines in Mucosal Applications
Natalija Van Braeckel-Budimir (2013)
Surface display on lactic acid bacteria without genetic modification: strategies and applications
Ruifeng Mao (2016)
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