In vivo interpretation of model predicted inhibition in acrylate pathway engineered Lactococcus lactis

2020 ◽  
Vol 117 (12) ◽  
pp. 3785-3798
Author(s):  
Sowmiya Balasubramanian ◽  
Priyadharshini Chandrasekran ◽  
Anitha J. R. Yesudhas ◽  
Padmapriya Ganapathyraman ◽  
Mark A. Eiteman ◽  
...  
Keyword(s):  
Lactococcus Lactis ◽  
Acrylate Pathway

scholarly journals In vivo interpretation of model predicted inhibition in acrylate pathway engineered Lactococcus lactis

Authorea ◽  
2020 ◽  
Author(s):  
Sowmiya Balasubramanian ◽  
Priyadharshini Chandrasekran ◽  
Vijayalaksmi Kandasamy ◽  
Mark Eiteman ◽  
Ramalingam Subramanian
Keyword(s):  
Lactococcus Lactis ◽  
Acrylate Pathway

Varying influence of the autolysin, N-acetyl muramidase, and the cell envelope proteinase on the rate of autolysis of six commercial Lactococcus lactis cheese starter bacteria grown in milk

2000 ◽  
Vol 67 (4) ◽  
pp. 585-596 ◽  
Author(s):  
SELVARANI GOVINDASAMY-LUCEY ◽  
PRAMOD K. GOPAL ◽  
PATRICK A. SULLIVAN ◽  
CHRISTOPHER J. PILLIDGE
Keyword(s):  
Lactococcus Lactis ◽  
Cell Walls ◽  
Cell Envelope ◽  
Laboratory Strain ◽  
Proteolytic Cleavage ◽  
Zymogram Analysis ◽  
Sds Page ◽  
Derivatives Of ◽  
Free Strains

The autolysin, N-acetyl muramidase (AcmA), of six commercial Lactococcus lactis subsp. cremoris starter strains and eight Lc. lactis subsp. cremoris derivatives or plasmid-free strains was shown by renaturing SDS-PAGE (zymogram analysis) to be degraded by the cell envelope proteinase (lactocepin; EC 3.4.21.96) after growth of strains in milk at 30 °C for 72 h. Degradation of AcmA was less in starter strains and derivatives producing lactocepin I/III (intermediate specificity) than in strains producing lactocepin I. This supports previous observations on AcmA degradation in derivatives of the laboratory strain Lc. lactis subsp. cremoris MG1363 (Buist et al. Journal of Bacteriology180 5947–5953 1998). In contrast to the MG1363 derivatives, however, the extent of autolysis in milk of the commercial Lc. lactis subsp. cremoris starter strains in this study did not always correlate with lactocepin specificity and AcmA degradation. The distribution of autolysins within the cell envelope of Lc. lactis subsp. cremoris starter strains and derivatives harvested during growth in milk was compared by zymogram analysis. AcmA was found associated with cell membranes as well as cell walls and some cleavage of AcmA occurred independently of lactocepin activity. An AcmA product intermediate in size between precursor (46 kDa) and mature (41 kDa) forms of AcmA was clearly visible on zymograms, even in the absence of lactocepin I activity. These results show that autolysis of commercial Lc. lactis subsp. cremoris starter strains is not primarily determined by AcmA activity in relation to lactocepin specificity and that proteolytic cleavage of AcmA in vivo is not fully defined.

The antibacterial activity expressed in vitro and in vivo by Lactococcus lactis subsp. lactis CNRZ 1427 against Salmonella sp.

2015 ◽  
Vol 4 (5) ◽  
pp. 240-246
Author(s):  
Menad Najett ◽  
Chougrani Fadelaa ◽  
Moghtet Snoussi ◽  
Cheriguene Abderrahim
Keyword(s):  
Antibacterial Activity ◽  
Lactococcus Lactis ◽  
Bile Salts ◽  
Therapeutic Agent ◽  
Lactococcus Lactis Subsp ◽  
Beneficial Effects ◽  
Gastrointestinal Conditions ◽  
Therapeutic Possibilities

  The present study focused on the beneficial effects of Lactococcus lactis subsp lactis CNRZ 1427 with possible use as a therapeutic agent against Sal-monella sp.; also we have proposed different therapeutic possibilities of our situation against a pathogen Salmonella sp. We have conducted two tests In Vitro and In Vivo; where it is noted that treatment in the presence of this lac-tic strain is effective since it causes a remarkable decrease of the pathogen agent. At the end, the effectiveness of this lactic strain was confirmed by testing for resistance to gastrointestinal conditions (pH, bile salts and diges-tive enzymes).

scholarly journals The Plasmodium falciparum circumsporozoite protein produced in Lactococcus lactis is pure and stable

2019 ◽  
Vol 295 (2) ◽  
pp. 403-414 ◽  
Author(s):  
Susheel K. Singh ◽  
Jordan Plieskatt ◽  
Bishwanath Kumar Chourasia ◽  
Vandana Singh ◽  
Judith M. Bolscher ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Lactococcus Lactis ◽  
Malaria Vaccine ◽  
Vaccine Development ◽  
Expression System ◽  
Surface Protein ◽  
Circumsporozoite Protein ◽  
Full Length

The Plasmodium falciparum circumsporozoite protein (PfCSP) is a sporozoite surface protein whose role in sporozoite motility and cell invasion has made it the leading candidate for a pre-erythrocytic malaria vaccine. However, production of high yields of soluble recombinant PfCSP, including its extensive NANP and NVDP repeats, has proven problematic. Here, we report on the development and characterization of a secreted, soluble, and stable full-length PfCSP (containing 4 NVDP and 38 NANP repeats) produced in the Lactococcus lactis expression system. The recombinant full-length PfCSP, denoted PfCSP4/38, was produced initially with a histidine tag and purified by a simple two-step procedure. Importantly, the recombinant PfCSP4/38 retained a conformational epitope for antibodies as confirmed by both in vivo and in vitro characterizations. We characterized this complex protein by HPLC, light scattering, MS analysis, differential scanning fluorimetry, CD, SDS-PAGE, and immunoblotting with conformation-dependent and -independent mAbs, which confirmed it to be both pure and soluble. Moreover, we found that the recombinant protein is stable at both frozen and elevated-temperature storage conditions. When we used L. lactis–derived PfCSP4/38 to immunize mice, it elicited high levels of functional antibodies that had the capacity to modify sporozoite motility in vitro. We concluded that the reported yield, purity, results of biophysical analyses, and stability of PfCSP4/38 warrant further consideration of using the L. lactis system for the production of circumsporozoite proteins for preclinical and clinical applications in malaria vaccine development.

Recombinant Lactococcus lactis NZ3900 expressing bioactive human FGF21 reduced body weight of Db/Db mice through the activity of brown adipose tissue

2020 ◽  
Vol 11 (1) ◽  
pp. 67-78 ◽  
Author(s):  
W.-Y. Cao ◽  
M. Dong ◽  
Z.-Y. Hu ◽  
J. Wu ◽  
Y.-C. Li ◽  
...  
Keyword(s):  
Body Weight ◽  
Antibiotic Resistance ◽  
Adipose Tissue ◽  
Oral Administration ◽  
Brown Adipose Tissue ◽  
Lactococcus Lactis ◽  
Metabolic Diseases ◽  
Brown Adipose ◽  
Human Fgf21

Fibroblast growth factor 21 (FGF21), a metabolism regulator, has an important effect on metabolic diseases, such as obesity and diabetes. It is also expressed in mice, and the murine source has high homology with human FGF21. Recently, it has been extensively studied and has become a potential drug target for the treatment of metabolic diseases. As it is a protein-based hormone, FGF21 cannot be easily and quickly absorbed into the blood through oral administration. Moreover, it has a 0-2 h half-life in vivo, as shown in a previous study, thus its efficacy lasts for a short period of time when used to treat metabolic diseases, limiting its clinical applications. To avoid these limitations, we used Lactococcus lactis, a food-grade bacterium, as the host to express FGF21. It could be used successfully for the expression and long-term effect of FGF21 in vivo. Instead of antibiotic resistance genes, the LacF gene was used as a selection marker in the NZ3900/PNZ8149 expression system, which is safe and could reduce the antibiotic resistance crisis. In this study, we a constructed human FGF21 expressing L. lactis strain and administered it to Db/Db mice by gavage. Compared with the control group, the body weight of mice in the experimental group was significantly reduced, and the overall homeostasis was improved in mice treated with human FGF21. Moreover, the activity of brown adipose tissue was enhanced. These results revealed that oral administration of FGF21 through heterologous expression in L. lactis appears to be an effective approach for its clinical application.

Protective effects of a novel probiotic strain, Lactococcus lactis ML2018, in colitis: in vivo and in vitro evidence

2019 ◽  
Vol 10 (2) ◽  
pp. 1132-1145 ◽  
Author(s):  
Meiling Liu ◽  
Xiuxia Zhang ◽  
Yunpeng Hao ◽  
Jinhua Ding ◽  
Jing Shen ◽  
...  
Keyword(s):  
Immune Responses ◽  
Lactococcus Lactis ◽  
Intestinal Microbiota ◽  
Inflammatory Bowel Diseases ◽  
Probiotic Strain ◽  
Protective Effects ◽  
Bowel Diseases ◽  
Inflammatory Bowel

Multiple articles have confirmed that an imbalance of the intestinal microbiota is closely related to aberrant immune responses of the intestines and to the pathogenesis of inflammatory bowel diseases (IBDs).

scholarly journals Intra- and Interspecies Conjugal Transfer of Tn916-Like Elements from Lactococcus lactis In Vitro and In Vivo

2009 ◽  
Vol 75 (19) ◽  
pp. 6352-6360 ◽  
Author(s):  
Joanna Boguslawska ◽  
Joanna Zycka-Krzesinska ◽  
Andrea Wilcks ◽  
Jacek Bardowski
Keyword(s):  
Antibiotic Resistance ◽  
Lactococcus Lactis ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Raw Milk ◽  
M Gene ◽  
Transfer Resistance

ABSTRACT Tetracycline-resistant Lactococcus lactis strains originally isolated from Polish raw milk were analyzed for the ability to transfer their antibiotic resistance genes in vitro, using filter mating experiments, and in vivo, using germfree rats. Four of six analyzed L. lactis isolates were able to transfer tetracycline resistance determinants in vitro to L. lactis Bu2-60, at frequencies ranging from 10−5 to 10−7 transconjugants per recipient. Three of these four strains could also transfer resistance in vitro to Enterococcus faecalis JH2-2, whereas no transfer to Bacillus subtilis YBE01, Pseudomonas putida KT2442, Agrobacterium tumefaciens UBAPF2, or Escherichia coli JE2571 was observed. Rats were initially inoculated with the recipient E. faecalis strain JH2-2, and after a week, the L. lactis IBB477 and IBB487 donor strains were introduced. The first transconjugants were detected in fecal samples 3 days after introduction of the donors. A subtherapeutic concentration of tetracycline did not have any significant effect on the number of transconjugants, but transconjugants were observed earlier in animals dosed with this antibiotic. Molecular analysis of in vivo transconjugants containing the tet(M) gene showed that this gene was identical to tet(M) localized on the conjugative transposon Tn916. Primer-specific PCR confirmed that the Tn916 transposon was complete in all analyzed transconjugants and donors. This is the first study showing in vivo transfer of a Tn916-like antibiotic resistance transposon from L. lactis to E. faecalis. These data suggest that in certain cases food lactococci might be involved in the spread of antibiotic resistance genes to other lactic acid bacteria.

Sign in / Sign up

Export Citation Format

Share Document