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Purification and characterization of �-d-xylosidase from Lactobacillus brevis grown on xylo-oligosaccharides.

Lyned, D. Lasrado and Muralikrishna, G. (2013) Purification and characterization of �-d-xylosidase from Lactobacillus brevis grown on xylo-oligosaccharides. Carbohydrate Polymers, 92. pp. 1978-1983.

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Abstract

In the recent years there has been a growing interest in the use of oligosaccharides as prebiotics to modulate gut microbiota with an aim to improve the gut health. Though xylo-oligosaccharides (XOS) have been increasingly used as prebiotics, information pertaining to the enzymes used by lactobacilli to degrade these substrates is scanty. Present investigation reports the purification and characterization of �-d-xylosidase from Lactobacillus brevis NCDC01 grown on XOS. Three sequential steps consisting of ultra-filtration, DEAE cellulose ion-exchange and Sephacryl S-100 gel filtration chromatographies were employed to purify the enzyme to apparent homogeneity and it was found to be monomeric on SDSPAGE with an apparent molecular mass of ∼58.0 kDa. The pH and temperature optima were 6.0 and 40 ◦C respectively. The enzyme remained stable over a pH range of 5.5–7.5 and up to 50 ◦C for 30 min. Under optimum pH and temperature with p-nitrophenyl �-d-xylopyranoside as a substrate, the enzyme exhibited a Km of 0.87 mM. The enzyme does not require any metal ion for activity or stability but is completely inhibited by Hg2+, Pb2+, p-chloromercuribenzoate (PCMB), oxalic acid and citric acid. This is perhaps the first report on the purification and characterization of �-d-xylosidase from Lactobacillus brevis NCDC01.

Item Type: Article
Uncontrolled Keywords: Xylooligosaccharides, Lactobacillus brevis NCDC01, �-d-Xylosidase, Prebiotics, Purification
Subjects: 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology
600 Technology > 08 Food technology > 16 Nutritive value > 05 Enzymes
Divisions: Dept. of Biochemistry
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 24 Jan 2013 05:59
Last Modified: 24 Jan 2013 05:59
URI: http://ir.cftri.com/id/eprint/11103

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