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<p>β-(1,4)-Mannobiose is a functional oligosaccharide with multiple prebiotic properties. The limitation of existing production technologies has hindered its further development and applications. In this study, we developed an efficient and scalable method combining a three-step enzymatic hydrolysis process with microbial purification to produce high-purity mannobiose from locust bean gum (LBG). The enzymatic degradation was carried out using β-mannanases rManA and rMan113A, along with α-galactosidase rGal27A from the alkaliphilic <i>Bacillus</i> N16-5, sequentially breaking down the substrate into galactose, mannose, and mannobiose. <i>Lactobacillus casei</i> LH23 was then employed to selectively metabolize mannose and galactose, ensuring the exclusive production of mannobiose. In a 250 mL reaction system containing 5% LBG, mannobiose was obtained at a concentration of 25.99&#xa0;g/L with a conversion efficiency of 51.98%, representing 98% of the total sugar yield. This study highlights the significance of understanding microbial polysaccharide utilization and engineering specialized enzymes to enhance enzymatic hydrolysis strategies for targeted oligosaccharide production.</p> Graphical Abstracts <p></p>

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Mannobiose Production Through Synergistic Enzymatic Hydrolysis of Locust Bean Gum by Mannanases from Alkaliphilic Bacillus sp. N16-5

  • 期刊论文

  • 发表日期：

    2025年11月3日

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  • Zhiyue Men,
  • Bingju Pan,
  • Xinke Liu,
  • Hening Zhang,
  • Wenting Liu,
  • Nan Wang,
  • Xuegang Luo,
  • Lang Rao,
  • Junxun Li,
  • Yajian Song

摘要

β-(1,4)-Mannobiose is a functional oligosaccharide with multiple prebiotic properties. The limitation of existing production technologies has hindered its further development and applications. In this study, we developed an efficient and scalable method combining a three-step enzymatic hydrolysis process with microbial purification to produce high-purity mannobiose from locust bean gum (LBG). The enzymatic degradation was carried out using β-mannanases rManA and rMan113A, along with α-galactosidase rGal27A from the alkaliphilic Bacillus N16-5, sequentially breaking down the substrate into galactose, mannose, and mannobiose. Lactobacillus casei LH23 was then employed to selectively metabolize mannose and galactose, ensuring the exclusive production of mannobiose. In a 250 mL reaction system containing 5% LBG, mannobiose was obtained at a concentration of 25.99 g/L with a conversion efficiency of 51.98%, representing 98% of the total sugar yield. This study highlights the significance of understanding microbial polysaccharide utilization and engineering specialized enzymes to enhance enzymatic hydrolysis strategies for targeted oligosaccharide production.

Graphical Abstracts