Abstract
This study focused on the characterization of mannanase from Bacillus circulans NT 6.7 for mannooligosaccharides (MOS) production. The enzyme from B. circulans NT 6.7 was produced using defatted copra meal as a carbon source. The mannanase was purified by ultrafiltration and column chromatography of Q-Sepharose. The purified protein (M1) was a dimeric protein with a 40 kDa subunit. The purified M1 exhibited optimum pH and temperature at pH 6.0 and 60 °C, respectively. It was activated by Mn2+, Mg2+, and Cu2+, and as inhibited by EDTA (45–65 %). The purified enzyme exhibited high specificity to beta-mannan: konjac (glucomannan), locust bean gum (galactomannan), ivory nut (mannan), guar gum (galactomannan) and defatted copra meal (galactomannan). The defatted copra meal could be hydrolyzed by purified M1 into mannooligosaccharides which promoted beneficial bacteria, especially Lactobacillus group, and inhibited pathogenic bacteria; Shigella dysenteria DMST 1511, Staphylococcus aureus TISTR 029, and Salmonella enterica serovar Enteritidis DMST 17368. Therefore, the mannanase from B. circulans NT 6.7 would be a novel source of enzymes for the mannooligosaccharides production as prebiotics.
Highlights
In 2004, “prebiotic” was defined as ‘‘selectively fermented ingredients that allow specific changes, both in the composition and/or activity in the gastrointestinal microbiota that confers benefits upon host well-being and health” (Gibson et al 2004)
Defatted copra meal can be used as a substrate for mannanase production and mannooligosaccharide preparation
The purified M1 was specified to mannan substrates; konjac mannan, locust bean gum, ivory nut, guar gum and defatted copra meal
Summary
In 2004, “prebiotic” was defined as ‘‘selectively fermented ingredients that allow specific changes, both in the composition and/or activity in the gastrointestinal microbiota that confers benefits upon host well-being and health” (Gibson et al 2004). Pathogenic bacteria (Clostridia bacteroides, Escherichia coli and Salmonella) produce toxins which can affect human gastrointestinal tracts, resulting in diarrhea, vomiting and nausea (Manning 2004). Some prebiotics, such as inulin, galactooligosaccharides (GOS), fructooligosaccharides (FOS), xylooligosaccharides (XOS), and mannooligosaccharides (MOS), are widely used in the market. Mannooligosaccharides are derived from the cell wall of the yeast Saccharomyces cerevisiae is commercially available as a feed supplement (Ferket et al 2002). They can be produced from plant mannan, such as konjac, ivory nut, locust bean, palm kernel, coffee bean, and copra meal. Mannan consists of repeating β-1, 4 mannose units and a few α-1, 6-galactose units attached to a β-1, 4 mannose backbone (Mohammad et al 1996)
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