Production and purification of mannan oligosaccharide with epithelial tight junction enhancing activity

PeerJ

article

Chatchai Nopvichai¹  Thanapon Charoenwongpaiboon¹  Navaporn Luengluepunya¹  Kazuo Ito²  Chatchai Muanprasat³  Rath Pichyangkura¹ 

[1] Department of Biochemistry, Faculty of Science, Chulalongkorn University;Graduate School of Science, Osaka City University;Department of Physiology, Faculty of Science, Mahidol University

关键词: Tight junction integration;    Mannan oligosaccharide;    Mannanase;   

DOI  :  10.7717/peerj.7206

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

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).

【 授权许可】

CC BY