Skip to main content
Log in

Antibiotic-free production of sucrose isomerase in Bacillus subtilis by genome integration

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript


Sucrose isomerase (SIase) catalyzes the hydrolysis and isomerization of sucrose to form isomaltulose, a valuable functional sugar widely used in the food industry. However, the lack of safe and efficient heterologous expression systems hinders SIase production and application. In this study, we achieved antibiotic-free SIase expression in Bacillus subtilis through genome integration. Using CRISPR/Cas9 system, SIase expression cassettes were integrated into various genomic loci, including amyE and ctc, both individually and in combination, resulting in single-copy and muti-copy integration strains. Engineered strains with a maltose-inducible promoter effectively expressed and secreted SIase. Notably, multi-copy strain exhibited enhanced SIase production, achieving 4.4 U/mL extracellular activity in shake flask cultivations. Furthermore, crude enzyme solution from engineered strain transformed high concentrations sucrose into high yields of isomaltulose, reaching a maximum yield of 94.6%. These findings demonstrate antibiotic-free SIase production in B. subtilis via genome integration, laying the foundation for its industrial production and application.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The data that support the findings of the present study are available from the corresponding author on reasonable request.


Download references


This research was funded by Scientific Research Fund of Liaoning Provincial Education Department (Grant Number LJKFZ20220213), Dalian Polytechnic University College Student Innovation and Entrepreneurship Training Program Project (Grant Number 202310152083), National Natural Science Foundation of China (Grant Number 32072160), and Key Research Project of the Education Department of Liaoning Province (Grant Number LJKZZ20220061).

Author information

Authors and Affiliations



Investigation, data curation, writing-original draft, Mingyu Li and Ming Xu; Investigation, data curation, Xinrui Bai, Xiang Wan, and Meng Zhao; Investigation, data curation, supervision, conceptualization, writing-review and editing, funding acquisition, Conggang Wang; writing-review and editing, Xianzhen Li, Xiaoyi Chen, and Fan Yang. All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Conggang Wang or Fan Yang.

Ethics declarations

Competing interest

The authors have no relevant financial or non-financial interests to disclose.

Ethical approval

This study does not contain any experiments involving human or animal subjects.

Consent to participate

Informed consent was obtained from all individual participants included in this study.

Consent for publication

The authors agreed to publish this paper in Biotechnology Letters.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 28 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, M., Xu, M., Bai, X. et al. Antibiotic-free production of sucrose isomerase in Bacillus subtilis by genome integration. Biotechnol Lett (2024).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: