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One-Step Bioprocess of Inulin to Product Inulo-Oligosaccharides Using Bacillus subtilis Secreting an Extracellular Endo-Inulinase

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Abstract

Inulo-oligosaccharides (IOSs), a novel food additive and health product, represent a promising alternative to antibiotics. As prebiotics, IOSs can be obtained from inulin by endo-inulinase-mediated hydrolysis. Nonetheless, enzymatic catalysis is not feasible industrially because of the required catalytic conditions and cost. In this study, a 2331-bp optimized gene inuQ (from Pseudomonas mucidolens) encoding endo-inulinase was cloned into shuttle vector PHY300PLK and transfected into Bacillus subtilis WB800-R, with the simultaneous deletion of gene sacC encoding levanase. The maximal IOS yield after hydrolysis of the crude extract of inulin was 67.84 ± 0.72 g/L for a recombinant strain with the signal peptide nprB from alkaline protease and promoter P43. The conversion rate reached 75.38%. For the major IOSs, the degree of polymerization was between 3 and 5. This study offers a simple and efficient one-step bioprocess for IOS production from inulin through secretion of an extracellular heterologous endo-inulinase by B. subtilis.

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Funding

This work was financially supported by the National Natural Science Foundation of China (Grant 21376203), the Promotion Program of Achievements in Scientific Research for Industrial Production of Higher Education of Jiangsu Province (Grant JHB2011-54), Qinglan Project of Higher Education of Jiangsu Province, Overseas Research & Training Program for University Prominent Young & Middle-aged Teachers and Presidents of Jiangsu Province, Jiangsu Agricultural Key Technology Research and Development Program (Grant BE2012394).

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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing Tech University, Nanjing, 211816, People’s Republic of China

    Ruifan Jiang, Yibin Qiu, Weiwei Huang & Hong Xu

  2. School of Marine and Bioengineering, Yancheng Institute of Technology, Yancheng, 224051, People’s Republic of China

    Ruifan Jiang, Li Zhang, Feng Xue, Hao Ni, Difen Mei & Jian Gao

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  1. Ruifan Jiang
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  2. Yibin Qiu
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  4. Li Zhang
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  7. Difen Mei
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  8. Jian Gao
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  9. Hong Xu
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Correspondence to Jian Gao or Hong Xu.

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Jiang, R., Qiu, Y., Huang, W. et al. One-Step Bioprocess of Inulin to Product Inulo-Oligosaccharides Using Bacillus subtilis Secreting an Extracellular Endo-Inulinase. Appl Biochem Biotechnol 187, 116–128 (2019). https://doi.org/10.1007/s12010-018-2806-3

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