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Identification of a Proteinaceous Component in the Leaf of Moringa Oleifera lam. with Effects on High Serum Creatinine.

Sahoo S, Raghavendra KM, Biswas S - Indian J Pharm Sci (2014)

Bottom Line: Apart from bactericidal effects, the parts of this plant have been effectively used in the treatment of circulatory, respiratory, endocrine, digestive as well as neural disorders.The crude extract was partially purified initially and eventually purified to completion as well.In light of the fact that increased level of serum creatinine levels have adverse downstream effects on the heart, lungs and other organs, this communication assumes significance because it suggests a way of reducing the level of serum creatinine as an emergency measure.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Pilani-K. K. Birla Goa Campus, Zuarinagar, Goa-403 726, India.

ABSTRACT
Moringa oleifera Lam. has been an important plant in the history of mankind, both for its nutritional and medicinal uses. Apart from bactericidal effects, the parts of this plant have been effectively used in the treatment of circulatory, respiratory, endocrine, digestive as well as neural disorders. Till date, though, there has been no reported activity of the involvement of any proteinaceous extract from M. oleifera on high levels of serum creatinine. To address this issue, blood samples with high levels of serum creatinine (2 mg/dl and above) were treated with leaf extract from M. oleifera. The crude extract was partially purified initially and eventually purified to completion as well. All these proteinaceous fractions were used to treat samples with high levels of serum creatinine as mentioned above. While the treatment of serum sample having high creatinine with crude extract and partially purified protein fractions showed a decrease of approximately 20% in the levels of serum creatinine over a period of 24 h, the samples treated with purified protein fraction reduced the serum creatinine level by 50%. In light of the fact that increased level of serum creatinine levels have adverse downstream effects on the heart, lungs and other organs, this communication assumes significance because it suggests a way of reducing the level of serum creatinine as an emergency measure. Further, the identification and characterisation of this proteinaceous component and possible in vivo experiments would provide a major tool for the treatment of downstream complications associated with increased serum creatinine via a new sources, albeit a natural one.

No MeSH data available.


SDS-PAGE profile of purification of crude extract of Moringa oleifera Lam. Lane 1: Crude extract, Lane 2: Ammonium sulphate pellet, Lane 3: NEX-GEN-PinkADD prestained protein ladder 10-175 kDa, Lane 4: Purified fraction after gel filtration, Lane 5: Purified fraction after ion exchange purification. The figures beside lane 3 indicate molecular weight in kDa.
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Figure 1: SDS-PAGE profile of purification of crude extract of Moringa oleifera Lam. Lane 1: Crude extract, Lane 2: Ammonium sulphate pellet, Lane 3: NEX-GEN-PinkADD prestained protein ladder 10-175 kDa, Lane 4: Purified fraction after gel filtration, Lane 5: Purified fraction after ion exchange purification. The figures beside lane 3 indicate molecular weight in kDa.

Mentions: The crude M. oleifera leaf extract, as prepared earlier, was estimated to contain 3 mg/ml of total protein. This crude extract was subjected to ammonium sulphate precipitation. It was standardised to achieve maximum separation of components at 50% saturation level. Both the supernatant and the resuspended pellet were dialysed overnight against extraction buffer separately and stored as fractions S (supernatant) and P (pellet) for further purification. S and P were separately differentiated into further fractions by gel filtration chromatography using a Superdex 200 prep grade column from GE Healthcare LifeSciences (Uppsala, Sweden). Checking for purity of samples, it was observed that one particular protein B65 (around 65 kDa) was the dominant protein in the pellet fractions. However, purification was not total, but only partial after the gel filtration step. Hence, the gel filtration fractions were subjected to further purification by ion-exchange chromatography using a HiTrap Q sepharose prepacked column from GE Healthcare LifeSciences (Uppsala, Sweden). The resulting fractions showed almost complete purification of the protein B65 (fig. 1).


Identification of a Proteinaceous Component in the Leaf of Moringa Oleifera lam. with Effects on High Serum Creatinine.

Sahoo S, Raghavendra KM, Biswas S - Indian J Pharm Sci (2014)

SDS-PAGE profile of purification of crude extract of Moringa oleifera Lam. Lane 1: Crude extract, Lane 2: Ammonium sulphate pellet, Lane 3: NEX-GEN-PinkADD prestained protein ladder 10-175 kDa, Lane 4: Purified fraction after gel filtration, Lane 5: Purified fraction after ion exchange purification. The figures beside lane 3 indicate molecular weight in kDa.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4007259&req=5

Figure 1: SDS-PAGE profile of purification of crude extract of Moringa oleifera Lam. Lane 1: Crude extract, Lane 2: Ammonium sulphate pellet, Lane 3: NEX-GEN-PinkADD prestained protein ladder 10-175 kDa, Lane 4: Purified fraction after gel filtration, Lane 5: Purified fraction after ion exchange purification. The figures beside lane 3 indicate molecular weight in kDa.
Mentions: The crude M. oleifera leaf extract, as prepared earlier, was estimated to contain 3 mg/ml of total protein. This crude extract was subjected to ammonium sulphate precipitation. It was standardised to achieve maximum separation of components at 50% saturation level. Both the supernatant and the resuspended pellet were dialysed overnight against extraction buffer separately and stored as fractions S (supernatant) and P (pellet) for further purification. S and P were separately differentiated into further fractions by gel filtration chromatography using a Superdex 200 prep grade column from GE Healthcare LifeSciences (Uppsala, Sweden). Checking for purity of samples, it was observed that one particular protein B65 (around 65 kDa) was the dominant protein in the pellet fractions. However, purification was not total, but only partial after the gel filtration step. Hence, the gel filtration fractions were subjected to further purification by ion-exchange chromatography using a HiTrap Q sepharose prepacked column from GE Healthcare LifeSciences (Uppsala, Sweden). The resulting fractions showed almost complete purification of the protein B65 (fig. 1).

Bottom Line: Apart from bactericidal effects, the parts of this plant have been effectively used in the treatment of circulatory, respiratory, endocrine, digestive as well as neural disorders.The crude extract was partially purified initially and eventually purified to completion as well.In light of the fact that increased level of serum creatinine levels have adverse downstream effects on the heart, lungs and other organs, this communication assumes significance because it suggests a way of reducing the level of serum creatinine as an emergency measure.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Pilani-K. K. Birla Goa Campus, Zuarinagar, Goa-403 726, India.

ABSTRACT
Moringa oleifera Lam. has been an important plant in the history of mankind, both for its nutritional and medicinal uses. Apart from bactericidal effects, the parts of this plant have been effectively used in the treatment of circulatory, respiratory, endocrine, digestive as well as neural disorders. Till date, though, there has been no reported activity of the involvement of any proteinaceous extract from M. oleifera on high levels of serum creatinine. To address this issue, blood samples with high levels of serum creatinine (2 mg/dl and above) were treated with leaf extract from M. oleifera. The crude extract was partially purified initially and eventually purified to completion as well. All these proteinaceous fractions were used to treat samples with high levels of serum creatinine as mentioned above. While the treatment of serum sample having high creatinine with crude extract and partially purified protein fractions showed a decrease of approximately 20% in the levels of serum creatinine over a period of 24 h, the samples treated with purified protein fraction reduced the serum creatinine level by 50%. In light of the fact that increased level of serum creatinine levels have adverse downstream effects on the heart, lungs and other organs, this communication assumes significance because it suggests a way of reducing the level of serum creatinine as an emergency measure. Further, the identification and characterisation of this proteinaceous component and possible in vivo experiments would provide a major tool for the treatment of downstream complications associated with increased serum creatinine via a new sources, albeit a natural one.

No MeSH data available.