Size Exclusion Chromatography Method for Purification of Nicotinamide Mononucleotide (NMN) from Bacterial Cells.
George Cătălin MarinescuRoua-Gabriela PopescuAnca DinischiotuPublished in: Scientific reports (2018)
Over 12% of the world's health resources are spent on treating diabetes, as high blood glucose is the third cause of mortality worldwide. Insulin resistance is the basis of the most common form of diabetes: type 2 diabetes. Recent animal studies report successful attempts at reversing type 2 diabetes by the administering of the NAD+ precursor nicotinamide mononucleotide (NMN). However, the current high price of this molecule urges for more efficient and cost-effective production methods. This work proposes a method for purifying NMN by Size Exclusion Chromatography (SEC) on silica with a covalently attached coating of poly(2-hydroxyethyl aspartamide) (PolyHEA) stationary phase using an isocratic elution with a denaturing mobile phase (50 mM formic acid) from a complex molecular mixture such as a fermentation broth. The eluted peaks were identified by UV-Vis analysis and confirmed with ESI+ mass spectrometry and a HPLC reversed-phase method. The proposed SEC method is simple, patent-free, directly applicable for industrial production with a minimum scale up effort. The need for multiple chromatographic steps is eliminated and the lysate filtration and clarification steps are simplified. Substantial reduction in NMN production costs and increased purity of NMN to the level suitable for usage in humans are expected.
Keyphrases
- type diabetes
- glycemic control
- mass spectrometry
- blood glucose
- liquid chromatography
- insulin resistance
- high performance liquid chromatography
- tandem mass spectrometry
- cardiovascular disease
- ms ms
- simultaneous determination
- healthcare
- induced apoptosis
- high speed
- gas chromatography
- capillary electrophoresis
- cardiovascular events
- weight loss
- public health
- high resolution
- cell cycle arrest
- high fat diet
- adipose tissue
- coronary artery disease
- heavy metals
- mental health
- skeletal muscle
- metabolic syndrome
- risk assessment
- recombinant human
- pi k akt
- social media
- blood pressure
- lactic acid