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Purification and characterization of an alkali-thermostable β-mannanase from Bacillus nealsonii PN-11 and its application in mannooligosaccharides preparation having prebiotic potential

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Abstract

An alkali-thermostable β-mannanase from Bacillus nealsonii PN-11 was purified 38.96-fold to homogeneity with specific activity of 2,288.90 ± 27.80 U mg−1 protein and final recovery of 8.92 ± 0.09 %. The purified β-mannanase was an extracellular monomeric protein with a molecular mass of 50 kDa on SDS–PAGE. The first 20 N-terminal amino acid sequence of mannanase enzyme was MVVKKLSSFILILLLVTSAL. The optimal temperature and pH for enzyme were 65 °C and 8.8, respectively. It was completely stable at 60 °C for 3 h and retained >50 ± 1.0 % activity at 70 °C up to 3 h. The β-mannanase was highly stable between pH 5–10 and retained >85 % of the initial activity for 3 h. The metal ions Ni+2, Co+2, Zn+2 and Mg+2 enhanced the enzyme activity. The enzyme remained stable after 3 h of preincubation with most of the tested organic solvents. According to substrate specificity study, the purified mannanase had high specificity to locust bean gum which was degraded mainly to mannooligosaccharides (MOS) like mannotriose, mannotetraose and mannopentose. These MOS enhanced the growth of Lactobacillus casei but inhibited the growth of Salmonella enterica indicating potential prebiotic properties. The properties of the purified β-mannanase from B. nealsonii PN-11 make this enzyme attractive for biotechnological applications.

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Acknowledgments

Prakram Singh Chauhan is thankful to Council of Scientific and Industrial Research (CSIR), New Delhi, India, for providing a Senior Research Fellowship.

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None.

Compliance with Ethics Requirements

All ethical considerations were complied with while working with bacterial strains.

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Authors and Affiliations

  1. Department of Microbiology, BMS Block, Panjab University, Sector-14, Chandigarh, 160014, India

    Prakram Singh Chauhan, Prince Sharma & Naveen Gupta

  2. Department of Microbiology, Guru Nanak Khalsa College, City Centre Road, Yamunanagar, 135001, Haryana, India

    Neena Puri

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  1. Prakram Singh Chauhan
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  2. Prince Sharma
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  4. Naveen Gupta
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Correspondence to Naveen Gupta.

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Chauhan, P.S., Sharma, P., Puri, N. et al. Purification and characterization of an alkali-thermostable β-mannanase from Bacillus nealsonii PN-11 and its application in mannooligosaccharides preparation having prebiotic potential. Eur Food Res Technol 238, 927–936 (2014). https://doi.org/10.1007/s00217-014-2170-7

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