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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

Representative 2D-DIGE fluorescent gel images of Lactobacillus acidophilus NCFM soluble cytosolic proteins for (a) the acidic region pH 4–7 and (b) the alkaline region pH 6–11. 2D-DIGE Coomassie Brilliant Blue gel images showing numbers indicating spots picked for identification by mass spectrometry are shown for (c) the acidic region pH 4–7 and (d) the alkaline region pH 6–11.
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fig1: Representative 2D-DIGE fluorescent gel images of Lactobacillus acidophilus NCFM soluble cytosolic proteins for (a) the acidic region pH 4–7 and (b) the alkaline region pH 6–11. 2D-DIGE Coomassie Brilliant Blue gel images showing numbers indicating spots picked for identification by mass spectrometry are shown for (c) the acidic region pH 4–7 and (d) the alkaline region pH 6–11.

Mentions: The present investigation of cytosolic proteomes of NCFM grown on cellobiose, a potential synbiotic combination, and glucose identified a total of 136 differentially abundant spots in 2D-DIGE (Figure 1). Of these 108 (spots 1–108) were in the acidic region (pH 4–7, Figure 1(c)) and 28 spots in the alkaline region (pH 6–11, Figure 1(d)), respectively. This difference is in accordance with the cytosolic proteome distribution of NCFM [8, 29]. Among the 136 spots, 81 showed higher (1.5–13.9-fold) and 55 lower (1.5–7.8-fold) relative abundance. Proteins were identified with confidence by MS and/or MS/MS in 81 of 136 picked spots, representing 49 unique proteins. The 81 spots correspond to 64 and 17 identifications from the acidic and alkaline regions, respectively (Tables S1 and S2). The hypothetical protein LBA0890 (spots 54–56 and 124) was identified both in the acidic and alkaline proteome in the overlapping region (pI 6-7).


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)

Representative 2D-DIGE fluorescent gel images of Lactobacillus acidophilus NCFM soluble cytosolic proteins for (a) the acidic region pH 4–7 and (b) the alkaline region pH 6–11. 2D-DIGE Coomassie Brilliant Blue gel images showing numbers indicating spots picked for identification by mass spectrometry are shown for (c) the acidic region pH 4–7 and (d) the alkaline region pH 6–11.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4417578&req=5

fig1: Representative 2D-DIGE fluorescent gel images of Lactobacillus acidophilus NCFM soluble cytosolic proteins for (a) the acidic region pH 4–7 and (b) the alkaline region pH 6–11. 2D-DIGE Coomassie Brilliant Blue gel images showing numbers indicating spots picked for identification by mass spectrometry are shown for (c) the acidic region pH 4–7 and (d) the alkaline region pH 6–11.
Mentions: The present investigation of cytosolic proteomes of NCFM grown on cellobiose, a potential synbiotic combination, and glucose identified a total of 136 differentially abundant spots in 2D-DIGE (Figure 1). Of these 108 (spots 1–108) were in the acidic region (pH 4–7, Figure 1(c)) and 28 spots in the alkaline region (pH 6–11, Figure 1(d)), respectively. This difference is in accordance with the cytosolic proteome distribution of NCFM [8, 29]. Among the 136 spots, 81 showed higher (1.5–13.9-fold) and 55 lower (1.5–7.8-fold) relative abundance. Proteins were identified with confidence by MS and/or MS/MS in 81 of 136 picked spots, representing 49 unique proteins. The 81 spots correspond to 64 and 17 identifications from the acidic and alkaline regions, respectively (Tables S1 and S2). The hypothetical protein LBA0890 (spots 54–56 and 124) was identified both in the acidic and alkaline proteome in the overlapping region (pI 6-7).

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