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Purification and characterization of [Formula: see text]-mannanase from Bacillus pumilus (M27) and its applications

Ahmet Adiguzel1, Hayrunnisa Nadaroglu2, Gulsah Adiguzel3

  • 1Faculty of Science, Department of Molecular Biology and Genetic, Ataturk University, 25240 Erzurum, Turkey.

Journal of Food Science and Technology
|August 6, 2015
PubMed

Insights

A novel thermo alkaline mannanase from Bacillus pumilus exhibits high stability across a broad pH and temperature range. This purified enzyme effectively clarifies fruit juices, particularly apple juice, demonstrating significant potential for industrial applications.

Area of Science:

  • Biochemistry
  • Enzymology
  • Food Science

Background:

  • Mannanase enzymes are crucial for breaking down mannans, complex carbohydrates found in plant cell walls.
  • Thermo alkaline mannanases are of particular interest due to their stability under harsh industrial processing conditions.
  • Bacillus pumilus is a known source of various industrial enzymes.

Purpose of the Study:

  • To purify and characterize a thermo alkaline mannanase from Bacillus pumilus (M27).
  • To evaluate the enzyme's stability under different pH and temperature conditions.
  • To assess the enzyme's efficacy in clarifying fruit juices.

Main Methods:

  • Enzyme purification using ammonium sulphate precipitation, DEAE-Sephadex ion exchange chromatography, and Sephacryl S200 gel filtration.
  • Enzyme characterization including subunit analysis via SDS-PAGE, determination of optimal pH and temperature, and stability assays.
  • Fruit juice clarification assays using purified mannanase on orange, apricot, grape, and apple juices.

Main Results:

  • A 111-fold purified mannanase with 36% yield was obtained.
  • The enzyme consists of two subunits (35 kDa and 55 kDa) and exhibits optimal activity at pH 8 and 60°C.
  • The mannanase retained activity over a wide pH range (3-11) and up to 90°C, with significant clarifying efficiency on apple juice (154%).

Conclusions:

  • The purified thermo alkaline mannanase from Bacillus pumilus is a robust enzyme with broad operational stability.
  • Its high clarifying efficiency on apple juice suggests potential applications in the food and beverage industry.
  • Further investigation into metal ion effects and industrial-scale application is warranted.
Keywords:
Bacillus pumilus (M27)CharacterizationClarificationMannanasePurification

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