New strategy for enhancement of microbial viability in simulated gastric conditions based on display of starch-binding domain on cell surface.
The C-terminal region of the peptidoglycan hydrolase (CPH) of Lactococcus lactis IL1403 fused to the linker region and the starch-binding domain (SBD) of the *-amylase of Streptococcus bovis 148 was produced intracellularly in Escherichia coli. The fusion protein (CPH-SBD) was able to bind to the cell surface of Lactobacillus casei NRRL B-441 and to corn starch. Therefore, adhesion of cells to corn starch was mediated by the fusion protein. At a cell density of 10(9) cfu/ml and a starch concentration of 5 mg/ml, CPH-SBD-displaying L. casei cells aggregated with corn starch, whereas the free cells of L. casei did not form any aggregates with corn starch. After incubation in simulated gastric juice (pH 3.0, 1 h), the survival percentages of free cells, amylose-coated free cells, and free cells mixed with corn starch were 0.074%, 7.2%, and 3.1% respectively. When CPH-SBD-displaying bacteria aggregated with corn starch, their survival percentage was 8% higher than that of free cells mixed with corn starch. The survival of the amylose-coated CPH-SBD-displaying L. casei cells was comparable to that of amylose-coated free cells, whereas the survival percentage of amylose-coated aggregates of CPH-SBD-displaying bacteria with corn starch was 28% higher than that of amylose-coated mixture of free cells with corn starch. These results demonstrate the potential usefulness of the cell-surface display technique for enhancement of the delivery of viable microorganisms to the intestinal tract.
Tarahomjoo
Shirin
S
Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
Katakura
Yoshio
Y
Shioya
Suteaki
S
D001665:Binding Sites; D032701:Biomimetics / Q000379:methods; D002448:Cell Adhesion; D002462:Cell Membrane / Q000378:metabolism / Q000382:microbiology*; D002470:Cell Survival; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005750:Gastric Juice / Q000382:microbiology*; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D019936:Probiotics; D013213:Starch / Q000378:metabolism*; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
D013213:Starch; D000516:alpha-Amylases; D009238:N-Acetylmuramoyl-L-alanine Amidase