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

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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.

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Acknowledgements

This work was supported by the Commission on Higher Education, the Ministry of Education, Royal Thai Government, and the ASEAN-EU University Network Programme (AUNP).

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Bangkok, Thailand

    Sudathip Titapoka, Suttipun Keawsompong & Sunee Nitisinprasert

  2. Department of Food Science and Technology, Division of Food Biotechnology, BOKU – University of Natural Resources and Applied Life Sciences, Vienna, Austria

    Dietmar Haltrich

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  1. Sudathip Titapoka
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  2. Suttipun Keawsompong
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  3. Dietmar Haltrich
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  4. Sunee Nitisinprasert
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Correspondence to Sunee Nitisinprasert.

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Titapoka, S., Keawsompong, S., Haltrich, D. et al. Selection and characterization of mannanase-producing bacteria useful for the formation of prebiotic manno-oligosaccharides from copra meal. World J Microbiol Biotechnol 24, 1425–1433 (2008). https://doi.org/10.1007/s11274-007-9627-9

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