Abstract
Lactobacillus rhamnosus GG (ATCC 53103) and Bifidobacterium longum NCC 2705 are among the most studied probiotics. However, the first evidence of acyl hydrolase/lipase of two annotated proteins, one in each genome of these strains, is reported in this work. Signal peptide analysis has predicted that these proteins are exported to the extracellular medium. Both proteins were produced in Escherichia coli, purified and characterized. Molecular masses (without signal peptides) were 27 and 52.3 kDa for the proteins of L. rhamnosus and B. longum, respectively. Asymmetrical flow field-flow fractionation analysis has shown that both proteins are present as monomers in their native forms at pH 7. Both have shown enzymatic activity on pNP-laurate at pH 7 and 37°C. The enzyme from L. rhamnosus was characterized deeper, showing preference on pNP-esters with short chain fatty acids. In addition, a computational model of the 3D structure has allowed the prediction of the catalytic amino acids. The enzymatic activities using synthetic substrates were very low for both enzymes. The investigation of natural substrates and biological functions of these enzymes is still open.
Highlights
Probiotics are microorganisms that provide beneficial effects for health and are gaining great interest both in the prevention as well as in the treatment of different diseases
Putative extracellular lipases were searched in the genomes of Lactobacillus rhamnosus, strain GG (ATCC 53103) and Bifidobacterium longum NCC 2705
The theoretical molecular weights of these truncated forms are 27058.74 Da (LrLyp) and 52360.25 Da (BlLyp) from L. rhamnosus and B. longum, respectively; which is consistent with the molecular mass determined by SDSPAGE (Figure 1)
Summary
Probiotics are microorganisms that provide beneficial effects for health and are gaining great interest both in the prevention as well as in the treatment of different diseases. The WHO (FAO/WHO) defines probiotic as “live microorganisms that, when administered in adequate amounts, confer a health benefit on the host” (Hill et al, 2014). Lactobacillus rhamnosus GG (ATCC 53103) and Bifidobacterium longum NCC 2705 are among the most studied probiotics. Several evidences support the positive effect of these strains against obesity, diabetes, inflammatory bowel disease, cancer, mental disorders, infections, diarrhea, immune disorders, systemic inflammation and others (Brusaferro et al, 2018; Consortium, 2018; Capurso, 2019). Acyl-Hydrolase/Lipase, Lactobacillus rhamnosus, Biffidobacterium longum in Google Patents gave more than 1250 and 1050 patents related with B. longum NCC 2705 and L. rhamnosus GG, respectively. L. rhamnosus is one of the most used bacteria in the probiotic food industry, which includes the manufacture of milkbased products such as yogurt-type drinks, and, probiotic fruit juices, berry soups and products based in fermented cereals and soy (Saxelin, 2008)
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