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An unusual GH1 β-glucosidase from marine sediment with β-galactosidase and transglycosidation activities for superior galacto-oligosaccharide synthesis

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Abstract

A novel β-glucosidase, BglD1 with high β-galactosidase and transglycosidation activities, was screened and cloned from the deep-sea bacterium Bacillus sp. D1. BglD1 exhibited the maximal β-glucosidase and β-galactosidase activities at 55–60 °C and pH 5.5–6.0. The enzyme maintained approximately 50% of its original activity at 35 °C and pH 6.0 after 120-h incubation. When applied to synthesize galacto-oligosaccharides (GOS), BglD1 generated 118.3 g/L GOS (33.8% (w/w)) from 350 g/L lactose, with trisaccharide Gal-β(1 → 3)-Lac and disaccharide Gal-β(1 → 4)-Gal as the main components. Furthermore, BglD1 could hydrolyze lactose in milk and produce GOS simultaneously. Using milk as the substrate, BglD1 hydrolyzed 88.5% lactose and produced 3.3 g/L GOS after incubation at 30 °C for 1 h. To improve the transglycosidation activity, a mutant BglD1:E224T was generated based on the semi-rational design. The GOS yield of BglD1:E224T was 11.5% higher than that of BglD1 when using lactose solution as the substrate. Thus, BglD1 and the mutant could be used as beneficial alternatives of the existing β-galactosidases for the production of GOS.

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Acknowledgments

The authors are grateful to Prof. Ursula Kües (University of Goettingen, Goettingen, Germany) for constructive suggestions.

Funding

This work was financially supported by the National Key Research and Development Program (grant number: 2018YFC0311106) and the Chinese National Natural Science Foundation (grant number: 31870056).

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Author notes

  1. Pengjun Deng and Chunyu Meng contributed equally to this work.

Authors and Affiliations

  1. School of Life Sciences, Anhui University, Hefei, 230601, Anhui, China

    Pengjun Deng, Chunyu Meng, Yan Wu, Jie Xu, Xuecheng Zhang, Yazhong Xiao, Zemin Fang & Wei Fang

  2. Anhui Key Laboratory of Modern Biomanufacturing, Hefei, 230601, Anhui, China

    Pengjun Deng, Chunyu Meng, Yan Wu, Jie Xu, Xuecheng Zhang, Yazhong Xiao, Zemin Fang & Wei Fang

  3. Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, 230601, Anhui, China

    Pengjun Deng, Chunyu Meng, Yan Wu, Jie Xu, Xuecheng Zhang, Yazhong Xiao, Zemin Fang & Wei Fang

  4. Department of Chemistry & Biochemistry, Florida International University, Miami, FL, 33199, USA

    Xiaoqing Tang & Xiaotang Wang

Authors
  1. Pengjun Deng
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  2. Chunyu Meng
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  3. Yan Wu
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  4. Jie Xu
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  7. Yazhong Xiao
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  8. Xiaotang Wang
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  9. Zemin Fang
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  10. Wei Fang
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Contributions

PD carried out the major experiments and helped to revise the manuscript. CM constructed the mutant of BglD1. YW screened the positive clones. JX and XT performed the docking analysis. XZ and YX performed the analysis of sequence and structure. XW and ZF polished the manuscript. WF wrote and edited the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Zemin Fang or Wei Fang.

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Highlights

1. BglD1 possessed high β-galactosidase and transglycosidation activities.

2. BglD1 synthesized GOS from high concentration lactose.

3. BglD1 hydrolyzed milk lactose and produced GOS simultaneously.

4. BglD1 was easy to be engineered for transglycosylation improvement.

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Deng, P., Meng, C., Wu, Y. et al. An unusual GH1 β-glucosidase from marine sediment with β-galactosidase and transglycosidation activities for superior galacto-oligosaccharide synthesis. Appl Microbiol Biotechnol 104, 4927–4943 (2020). https://doi.org/10.1007/s00253-020-10578-z

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