Abstract
BackgroundMannanan oligosaccharide (MOS) is well-known as effective supplement food for livestock to increase their nutrients absorption and health status, but the structure and identification of bioactive MOS remain unclear. In this study, MOS production was accomplished, using enzymatic hydrolysis of pretreated coconut meal substrate with recombinant mannanase.MethodsThe mannanase gene was cloned from Bacillus subtilis cAE24, then expressed in BL21. Purified Mannanase exhibit stability over a wide range of pH and temperature from pH 6–8 and 4 °C to 70 °C, respectively. SEM analysis revealed that sonication could change the surface characteristic of copra meal, which gave better MOS yield, compared to untreated substrates. The separation and purification of each MOS were achieved using Biogel-P2 column chromatography. Determination of biological active MOS species was also investigated. T84 cells were cultured and treated with each of the purified MOS species to determine their tight junction enhancing activity.ResultsScanning electron microscope imaging showed that pretreatment using sonication could disrupt the surface of copra meal better than grinding alone, which can improve the production of MOS. Pentamer of MOS (M5) significantly increased tight junction integration of T84 cells measured with TEER (p < 0.0001).
Highlights
Mannan oligosaccharide (MOS) is an indigestible short chain polymer and a well-known supplement for increasing the life quality of pets and livestock
We reported the production of Mannanan oligosaccharide (MOS) from pretreated copra meal by enzymatic hydrolysis using recombinant endo-1,4-beta mannosidase derived from Bacillus subtilis c AE24
The amino acid sequence of RMase24 was blasted with NCBI database and aligned with mannanase sequences from various Bacillus subtilis strain
Summary
Mannan oligosaccharide (MOS) is an indigestible short chain polymer and a well-known supplement for increasing the life quality of pets and livestock. Several reports revealed that MOS supplement could improve growth performance and body weight in various animals (Ai et al, 2011; Dimitroglou et al, 2010; Genc et al, 2007; Mansour et al, 2012; Staykov et al, 2007). Many studies reported that feeding MOS to rabbits could increase length, density and improve the organisation of the ileac villi. This suggested a higher rate of intestinal nutrient uptake; improved the growth performance (Mourão et al, 2006). Pentamer of MOS (M5) significantly increased tight junction integration of T84 cells measured with TEER (p < 0.0001)
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