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The Differential Proteome of the Probiotic Lactobacillus acidophilus NCFM Grown on the Potential Prebiotic Cellobiose Shows Upregulation of Two β -Glycoside Hydrolases.

van Zanten GC, Sparding N, Majumder A, Lahtinen SJ, Svensson B, Jacobsen S - Biomed Res Int (2015)

Bottom Line: Probiotics, prebiotics, and combinations thereof, that is, synbiotics, are known to exert beneficial health effects in humans; however interactions between pro- and prebiotics remain poorly understood at the molecular level.Many of these proteins were associated with energy metabolism, including the cellobiose related glycoside hydrolases phospho-β-glucosidase (LBA0881) and phospho-β-galactosidase II (LBA0726).Several of the upregulated or downregulated identified proteins associated with utilization of cellobiose indicate the presence of carbon catabolite repression and regulation of enzymes involved in carbohydrate metabolism.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg C, Denmark ; Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Søltofts Plads, Building 224, 2800 Kongens Lyngb, Denmark.

ABSTRACT
Probiotics, prebiotics, and combinations thereof, that is, synbiotics, are known to exert beneficial health effects in humans; however interactions between pro- and prebiotics remain poorly understood at the molecular level. The present study describes changes in abundance of different proteins of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) when grown on the potential prebiotic cellobiose as compared to glucose. Cytosolic cell extract proteomes after harvest at late exponential phase of NCFM grown on cellobiose or glucose were analyzed by two dimensional difference gel electrophoresis (2D-DIGE) in the acidic (pH 4-7) and the alkaline (pH 6-11) regions showing a total of 136 spots to change in abundance. Proteins were identified by MS or MS/MS from 81 of these spots representing 49 unique proteins and either increasing 1.5-13.9-fold or decreasing 1.5-7.8-fold in relative abundance. Many of these proteins were associated with energy metabolism, including the cellobiose related glycoside hydrolases phospho-β-glucosidase (LBA0881) and phospho-β-galactosidase II (LBA0726). The data provide insight into the utilization of the candidate prebiotic cellobiose by the probiotic bacterium NCFM. Several of the upregulated or downregulated identified proteins associated with utilization of cellobiose indicate the presence of carbon catabolite repression and regulation of enzymes involved in carbohydrate metabolism.

No MeSH data available.


Related in: MedlinePlus

(a) Schematic presentation of cellobiose entry and hydrolysis by L. acidophilus NCFM. Superscript letters T and P indicate upregulation by transcriptomics [19] or increase in abundance by proteomics, respectively (LBA0725: PTS II, LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, LBA0881: phospho-β-glucosidase, LBA0884: PTS IIC LBA0726: phospho-β-galactosidase II, and LBA0885: β-glucosidase). (b) Schematic presentation of gene clusters encoding glycoside hydrolases (LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0881: phospho-β-glucosidase, and LBA0885: β-glucosidase), shown as light grey arrows and PTSs (LBA0725: PTS II, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, and LBA0884: PTS IIC), shown as dark grey arrows, predicted to be cellobiose specific. Transcription antiterminator (LBA0724), hypothetical proteins (LBA0878, LBA0880, and LBA0883), and transcriptional regulators (LBA0875, LBA0882, and LBA0886) are shown as white arrows.
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fig3: (a) Schematic presentation of cellobiose entry and hydrolysis by L. acidophilus NCFM. Superscript letters T and P indicate upregulation by transcriptomics [19] or increase in abundance by proteomics, respectively (LBA0725: PTS II, LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, LBA0881: phospho-β-glucosidase, LBA0884: PTS IIC LBA0726: phospho-β-galactosidase II, and LBA0885: β-glucosidase). (b) Schematic presentation of gene clusters encoding glycoside hydrolases (LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0881: phospho-β-glucosidase, and LBA0885: β-glucosidase), shown as light grey arrows and PTSs (LBA0725: PTS II, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, and LBA0884: PTS IIC), shown as dark grey arrows, predicted to be cellobiose specific. Transcription antiterminator (LBA0724), hypothetical proteins (LBA0878, LBA0880, and LBA0883), and transcriptional regulators (LBA0875, LBA0882, and LBA0886) are shown as white arrows.

Mentions: Growth of NCFM on cellobiose elicited an increase in abundance of catabolite control protein A (CcpA; Table 1, spot 47). CcpA is a key enzyme in carbon catabolite repression, a mechanism that regulates enzymes involved in carbohydrate metabolism. Thus in the presence of preferred sugars (e.g., glucose) enzymes involved in utilization of less favorable sugars are repressed [34]. Carbohydrate phosphotransferase systems (PTSs) involved in uptake and phosphorylation of carbohydrates participate in the carbon catabolite repression [34]. When NCFM was grown on cellobiose, notably 2.4-fold upregulation was seen for phospho-β-glucosidase (LBA0881; Table 1, spot 17) located in a gene cluster with multiple PTSs predicted to be cellobiose specific (Figure 3). Interestingly a 7.0–7.4-fold increased abundance was observed for phospho-β-galactosidase II (LBA0726; Table 1, spots 16 and 18) encoded in a gene cluster together with another PTS (LBA0725; Figure 3) indicating a role in cellobiose utilization (Figure 3). This PTS LBA0725 has been reported to have 62% amino acid sequence identity to a PTS (ORF 1669) of L. gasseri 33323 of which expression was strongly induced by cellobiose [35]. In another low GC ratio, Gram-positive bacteria, Clostridium acetobutylicum, transcriptional analysis demonstrated that cellobiose induced expression of two PTSs and three glycoside hydrolases [25]. Transcriptional analysis of NCFM grown on cellobiose has reported upregulation of several genes within the loci LBA0724–LBA0726 and LBA0877–LBA0884 [26]. Taken together, the results of transcriptional and proteomics analyses indicate that NCFM contains more than one operon encoding protein involved in uptake and metabolism of cellobiose.


The Differential Proteome of the Probiotic Lactobacillus acidophilus NCFM Grown on the Potential Prebiotic Cellobiose Shows Upregulation of Two β -Glycoside Hydrolases.

van Zanten GC, Sparding N, Majumder A, Lahtinen SJ, Svensson B, Jacobsen S - Biomed Res Int (2015)

(a) Schematic presentation of cellobiose entry and hydrolysis by L. acidophilus NCFM. Superscript letters T and P indicate upregulation by transcriptomics [19] or increase in abundance by proteomics, respectively (LBA0725: PTS II, LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, LBA0881: phospho-β-glucosidase, LBA0884: PTS IIC LBA0726: phospho-β-galactosidase II, and LBA0885: β-glucosidase). (b) Schematic presentation of gene clusters encoding glycoside hydrolases (LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0881: phospho-β-glucosidase, and LBA0885: β-glucosidase), shown as light grey arrows and PTSs (LBA0725: PTS II, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, and LBA0884: PTS IIC), shown as dark grey arrows, predicted to be cellobiose specific. Transcription antiterminator (LBA0724), hypothetical proteins (LBA0878, LBA0880, and LBA0883), and transcriptional regulators (LBA0875, LBA0882, and LBA0886) are shown as white arrows.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: (a) Schematic presentation of cellobiose entry and hydrolysis by L. acidophilus NCFM. Superscript letters T and P indicate upregulation by transcriptomics [19] or increase in abundance by proteomics, respectively (LBA0725: PTS II, LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, LBA0881: phospho-β-glucosidase, LBA0884: PTS IIC LBA0726: phospho-β-galactosidase II, and LBA0885: β-glucosidase). (b) Schematic presentation of gene clusters encoding glycoside hydrolases (LBA0726: phospho-β-galactosidase II, LBA0874: phospho-β-galactosidase I, LBA0881: phospho-β-glucosidase, and LBA0885: β-glucosidase), shown as light grey arrows and PTSs (LBA0725: PTS II, LBA0876: PTS IIC, LBA0877: PTS IIA, LBA0879: PTS IIC, and LBA0884: PTS IIC), shown as dark grey arrows, predicted to be cellobiose specific. Transcription antiterminator (LBA0724), hypothetical proteins (LBA0878, LBA0880, and LBA0883), and transcriptional regulators (LBA0875, LBA0882, and LBA0886) are shown as white arrows.
Mentions: Growth of NCFM on cellobiose elicited an increase in abundance of catabolite control protein A (CcpA; Table 1, spot 47). CcpA is a key enzyme in carbon catabolite repression, a mechanism that regulates enzymes involved in carbohydrate metabolism. Thus in the presence of preferred sugars (e.g., glucose) enzymes involved in utilization of less favorable sugars are repressed [34]. Carbohydrate phosphotransferase systems (PTSs) involved in uptake and phosphorylation of carbohydrates participate in the carbon catabolite repression [34]. When NCFM was grown on cellobiose, notably 2.4-fold upregulation was seen for phospho-β-glucosidase (LBA0881; Table 1, spot 17) located in a gene cluster with multiple PTSs predicted to be cellobiose specific (Figure 3). Interestingly a 7.0–7.4-fold increased abundance was observed for phospho-β-galactosidase II (LBA0726; Table 1, spots 16 and 18) encoded in a gene cluster together with another PTS (LBA0725; Figure 3) indicating a role in cellobiose utilization (Figure 3). This PTS LBA0725 has been reported to have 62% amino acid sequence identity to a PTS (ORF 1669) of L. gasseri 33323 of which expression was strongly induced by cellobiose [35]. In another low GC ratio, Gram-positive bacteria, Clostridium acetobutylicum, transcriptional analysis demonstrated that cellobiose induced expression of two PTSs and three glycoside hydrolases [25]. Transcriptional analysis of NCFM grown on cellobiose has reported upregulation of several genes within the loci LBA0724–LBA0726 and LBA0877–LBA0884 [26]. Taken together, the results of transcriptional and proteomics analyses indicate that NCFM contains more than one operon encoding protein involved in uptake and metabolism of cellobiose.

Bottom Line: Probiotics, prebiotics, and combinations thereof, that is, synbiotics, are known to exert beneficial health effects in humans; however interactions between pro- and prebiotics remain poorly understood at the molecular level.Many of these proteins were associated with energy metabolism, including the cellobiose related glycoside hydrolases phospho-β-glucosidase (LBA0881) and phospho-β-galactosidase II (LBA0726).Several of the upregulated or downregulated identified proteins associated with utilization of cellobiose indicate the presence of carbon catabolite repression and regulation of enzymes involved in carbohydrate metabolism.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg C, Denmark ; Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, Søltofts Plads, Building 224, 2800 Kongens Lyngb, Denmark.

ABSTRACT
Probiotics, prebiotics, and combinations thereof, that is, synbiotics, are known to exert beneficial health effects in humans; however interactions between pro- and prebiotics remain poorly understood at the molecular level. The present study describes changes in abundance of different proteins of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) when grown on the potential prebiotic cellobiose as compared to glucose. Cytosolic cell extract proteomes after harvest at late exponential phase of NCFM grown on cellobiose or glucose were analyzed by two dimensional difference gel electrophoresis (2D-DIGE) in the acidic (pH 4-7) and the alkaline (pH 6-11) regions showing a total of 136 spots to change in abundance. Proteins were identified by MS or MS/MS from 81 of these spots representing 49 unique proteins and either increasing 1.5-13.9-fold or decreasing 1.5-7.8-fold in relative abundance. Many of these proteins were associated with energy metabolism, including the cellobiose related glycoside hydrolases phospho-β-glucosidase (LBA0881) and phospho-β-galactosidase II (LBA0726). The data provide insight into the utilization of the candidate prebiotic cellobiose by the probiotic bacterium NCFM. Several of the upregulated or downregulated identified proteins associated with utilization of cellobiose indicate the presence of carbon catabolite repression and regulation of enzymes involved in carbohydrate metabolism.

No MeSH data available.


Related in: MedlinePlus