Selection and characterization of mannanase-producing bacteria useful for the formation of prebiotic manno-oligosaccharides from copra meal

Abstract

In order to select bacterial strains effectively secreting mannanase activity for the production of prebiotic mannooligosaccharides, a two-step screening procedure was performed. Enriched cultures from isolation medium containing copra meal were primary screened on an isolation agar medium containing 1% locust bean gum (LBG), which resulted in 48 mannanase-producing bacterial isolates with significant clearing zones on the mannan-containing agar. However, only nine isolates showed appreciable mannanase activities against copra meal in their culture supernatants (0.054-0.185 U/mg of protein) as determined in a standard assay based on the detection of reducing sugars released from this substrate. The isolates CW2-3 and ST1-1 displayed the highest activity against LBG and copra meal, respectively. Copra mannan hydrolysates that were obtained by using crude mannanase from these nine isolates were further used for a secondary screening towards a growth-enhancing activity on Lactobacillus reuteri and inhibitory activity against Escherichia coli as well as Salmonella Enteritidis, resulting in 0.09-2.15 log CFU/ml enhancing activity and low inhibitory activity of 0.46-1.78 log CFU/ml as well as 0.37-1.72 log CFU/ml, respectively. The hydrolysate of CW2-3 mannanase showed the highest enhancing activity of 2.15 log CFU/ml while isolate ST1-1 was most effective with respect to growth inhibition against E. coli E010 and S. Enteritidis S003 with 0.76 and 1.61 log CFU/ml, respectively. Based on morphological, physical, biochemical and genetics properties, isolates CW2-3 and ST1-1 were identified as Klebsiella oxytoca and Acinetobacter sp., respectively. Crude mannanase activity from these two strains was characterized preliminarily. The pH optima of mannanase activity from Klebsiella oxytoca CW2-3 and Acinetobacter sp. ST1-1 were 7 and 6, respectively. The enzymes were stable at 4°C over a pH range of 3-6 and 3-10, respectively.

Application of Micro-Encapsulated Probiotic Bacillus NP5 and Prebiotic Mannan Oligosaccharide (MOS) to Prevent Streptococcosis on Tilapia Oreochromis niloticus

“…While, prebiotic is capable of giving beneficial effects to the host by modulation of beneficial bacteria selectively in the gastrointestinal tract of tilapia. According to Titapoka et al (2008), beneficial bacteria which have mannanase can utilize mannan oligosaccharide for growth and bacteria replication which contributes to the growth and digestibility of tilapia through exogenous digestive enzyme secretion. In addition, prebiotics are known can inhibit the adhesion of pathogenic bacteria in the intestine, improve growth, feed efficiency and composition of beneficial micro-flora in the intestine (Mahious et al, 2006).…”

Characterization of mannanase S1 from Klebsiella oxytoca KUB-CW2-3 and its application in copra mannan hydrolysis

“…Its β-mannanase yields were lower than those reported for some other bacterial and fungal sources [11][12][13] . However, copra hydrolysates produced by K. oxytoca KUB-CW2-3 clearly showed growth promoting properties for Lactobacillus reuteri but growth inhibitory effects for Salmonella Enteritidis growth, indicating valuable prebiotic properties of this copra hydrolysates 10 . So far, there has not been report on the property of the native enzyme and the action of mannan endo-1,4-β-mannosidase from K. oxytoca KUB-CW2-3.…”

“…oxytoca KUB-CW2-3 10 was cultivated in nutrient broth (NB; Pronadisa, Madrid, Spain) at 37°C for 12-15 h with shaking at 150 rpm. L. reuteri KUB-AC5, Escherichia coli E010, and Salmonella serovar Enteritidis S003 obtained from chicken intestine 14 were used as target strains to determine prebiotic properties of the oligosaccharide preparations.…”

“…These mannan endo-1,4-β-mannosidases produced by various species of bacteria, fungi, plants, and invertebrates have also a number of other applications in different sectors of food, feed, pharmaceutical, and pulp/paper industries 8 . Klebsiella oxytoca KUB-CW2-3 was isolated from fermented coconut waste and was shown to actively hydrolyse both locust bean gum and copra mannan 10 . Its β-mannanase yields were lower than those reported for some other bacterial and fungal sources [11][12][13] .…”

Comparative study between free and immobilized Penicillium chrysogenum mannanase: a local fungal isolate

“…As a trail, the resulting MOS were examined for their benefit effect for growing of lactobacteria as a specific nutrient. The aim of this study is investigating the immobilization of mannanase and to evaluate its biological activity as prebiotic agent for hydrolysate MOS productions (Tomotari and Indus 1990;Titapoka et al, 2008;Quigley, 2010;Gibson et al, 2004;Yopi et al, 2006;Ariandi et al, 2015) and efficiency of hydrolysis.…”