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The Construction of a Probiotic Lactic Acid Bacterium Expressing Acid-Resistant Phytase Enzyme

Year: 2015 Volume: 9 Issue: 8 963 - 968 Pages

Abstract: The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.

The Impact of the Cell-Free Solution of Lactic Acid Bacteria on Cadaverine Production by Listeria monocytogenes and Staphylococcus aureus in Lysine-Decarboxylase Broth

Year: 2015 Volume: 9 Issue: 3 309 - 317 Pages

Abstract: The influences of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on cadaverine and other biogenic amines production by Listeria monocytogenes and Staphylococcus aureus were investigated in lysine decarboxylase broth (LDB) using HPLC. Cell free solutions were prepared from Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus acidilactici and Streptococcus thermophiles. Two different concentrations that were 50% and 25% CFS and the control without CFSs were prepared. Significant variations on biogenic amine production were observed in the presence of L. monocytogenes and S. aureus (P < 0.05). The function of CFS on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine formation by L. monocytogenes and S. aureus in control were 500.9 and 948.1 mg/L, respectively while the CFSs of LAB induced 4-fold lower cadaverine production by L. monocytogenes and 7-fold lower cadaverine production by S. aureus. The CFSs resulted in strong decreases in cadaverine and putrescine production by L. monocytogenes and S. aureus, although remarkable increases were observed for histamine, spermidine, spermine, serotonin, dopamine, tyramine and agmatine in the presence of LAB in lysine decarboxylase broth.

Utilization of Sugarcane Bagasses for Lactic Acid Production by acid Hydrolysis and Fermentation using Lactobacillus sp

Year: 2012 Volume: 6 Issue: 6 510 - 515 Pages

Abstract: Sugarcane bagasses are one of the most extensively used agricultural residues. Using acid hydrolysis and fermentation, conversion of sugarcane bagasses to lactic acid was technically and economically feasible. This research was concerned with the solubility of lignin in ammonium hydroxide, acid hydrolysis and lactic acid fermentation by Lactococcus lactis, Lactobacillus delbrueckii, Lactobacillus plantarum, and Lactobacillus casei. The lignin extraction results for different ammonium hydroxide concentrations showed that 10 % (v/v) NH4OH was favorable to lignin dissolution. Acid hydrolysis can be enhanced with increasing acid concentration and reaction temperature. The optimum glucose and xylose concentrations occurred at 121 ○C for 1 hour hydrolysis time in 10% sulphuric acid solution were 32 and 11 g/l, respectively. In order to investigate the significance of medium composition on lactic acid production, experiments were undertaken whereby a culture of Lactococcus lactis was grown under various glucose, peptone, yeast extract and xylose concentrations. The optimum medium was composed of 5 g/l glucose, 2.5 g/l xylose, 10 g/l peptone and 5 g/l yeast extract. Lactococcus lactis represents the most efficient for lactic acid production amongst those considered. The lactic acid fermentation by Lactococcus lactis after 72 hours gave the highest yield of 1.4 (g lactic acid per g reducing sugar).