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Enhanced levansucrase expression in Bacillus subtilis via regulatory element optimization for high-yield lactosucrose production from sucrose and lactose.

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Title: Enhanced levansucrase expression in Bacillus subtilis via regulatory element optimization for high-yield lactosucrose production from sucrose and lactose.
Authors: He H; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China., Yang Y; Beijing Vocational College of Agriculture, Department of Food and Bioengineering, Beijing 102442, China., Wei Q; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China., Liu X; Shandong Haizhibao Ocean Technology Co., Ltd, Weihai, Shandong 264333, China., Xu W; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China., Zhang W; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China., Ni D; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Shandong Haizhibao Ocean Technology Co., Ltd, Weihai, Shandong 264333, China. Electronic address: daweini@jiangnan.edu.cn., Mu W; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
Source: Bioresource technology [Bioresour Technol] 2026 Sep; Vol. 456, pp. 134954. Date of Electronic Publication: 2026 May 22.
Publication Type: Journal Article
Language: English
Journal Info: Publisher: Elsevier Applied Science Country of Publication: England NLM ID: 9889523 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-2976 (Electronic) Linking ISSN: 09608524 NLM ISO Abbreviation: Bioresour Technol Subsets: MEDLINE
Imprint Name(s): Original Publication: Barking, Essex, England : New York, N.Y. : Elsevier Applied Science ; Elsevier Science Pub. Co., 1991-
MeSH Terms: Bacillus subtilis*/genetics , Bacillus subtilis*/metabolism , Bacillus subtilis*/enzymology , Hexosyltransferases*/metabolism , Hexosyltransferases*/genetics , Sucrose*/metabolism , Trisaccharides*/biosynthesis , Lactose*/metabolism , Regulatory Sequences, Nucleic Acid*/genetics, Hydrogen-Ion Concentration ; Gene Expression Regulation, Bacterial ; Fermentation ; Temperature ; Promoter Regions, Genetic
Abstract: Lactosucrose is a functional oligosaccharide with prebiotic properties and broad applications in the food and nutraceutical industries. In this study, a systematic strategy was developed to enhance lactosucrose production by optimizing levansucrase expression in Bacillus subtilis. Six levansucrases were screened, and levansucrase from Brenneria goodwinii (BgLS) exhibited the highest solubility and whole-cell catalytic activity. Whole-cell reaction conditions were optimized, identifying pH 6.0 and 50 °C as optimal. BgLS expression was further enhanced through translational regulation by engineering synthetic 5'-untranslated regions (5'-UTRs) and N-terminal coding sequences (NCSs), followed by combinatorial optimization. Subsequent transcriptional optimization using endogenous promoters further increased enzyme expression. The best engineered strain achieved a fermentation activity of 190.7 U/mL and produced lactosucrose with a 46.5% conversion rate from 200 g/L lactose and 300 g/L sucrose. This work demonstrates an efficient platform for high-level lactosucrose biosynthesis and highlights the effectiveness of multi-level gene expression regulation.
(Copyright © 2026 Elsevier Ltd. All rights reserved.)
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Contributed Indexing: Keywords: Bacillus subtilis; Lactosucrose; Levansucrase; Translational and transcriptional regulation; Whole-cellbiocatalysis
Substance Nomenclature: EC 2.4.1.10 (levansucrase)
EC 2.4.1.- (Hexosyltransferases)
57-50-1 (Sucrose)
0 (Trisaccharides)
J2B2A4N98G (Lactose)
Entry Date(s): Date Created: 20260523 Date Completed: 20260609 Latest Revision: 20260609
Update Code: 20260610
DOI: 10.1016/j.biortech.2026.134954
PMID: 42176820
Database: MEDLINE
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  Data: Enhanced levansucrase expression in Bacillus subtilis via regulatory element optimization for high-yield lactosucrose production from sucrose and lactose.
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  Data: <searchLink fieldCode="AU" term="%22He+H%22">He H</searchLink>; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.<br /><searchLink fieldCode="AU" term="%22Yang+Y%22">Yang Y</searchLink>; Beijing Vocational College of Agriculture, Department of Food and Bioengineering, Beijing 102442, China.<br /><searchLink fieldCode="AU" term="%22Wei+Q%22">Wei Q</searchLink>; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.<br /><searchLink fieldCode="AU" term="%22Liu+X%22">Liu X</searchLink>; Shandong Haizhibao Ocean Technology Co., Ltd, Weihai, Shandong 264333, China.<br /><searchLink fieldCode="AU" term="%22Xu+W%22">Xu W</searchLink>; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.<br /><searchLink fieldCode="AU" term="%22Zhang+W%22">Zhang W</searchLink>; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.<br /><searchLink fieldCode="AU" term="%22Ni+D%22">Ni D</searchLink>; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Shandong Haizhibao Ocean Technology Co., Ltd, Weihai, Shandong 264333, China. Electronic address: daweini@jiangnan.edu.cn.<br /><searchLink fieldCode="AU" term="%22Mu+W%22">Mu W</searchLink>; State Key Laboratory of Food Science and Resources, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
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  Data: <i>Publisher: </i><searchLink fieldCode="PB" term="%22Elsevier+Applied+Science%22">Elsevier Applied Science </searchLink><i>Country of Publication: </i>England <i>NLM ID: </i>9889523 <i>Publication Model: </i>Print-Electronic <i>Cited Medium: </i>Internet <i>ISSN: </i>1873-2976 (Electronic) <i>Linking ISSN: </i><searchLink fieldCode="IS" term="%2209608524%22">09608524 </searchLink><i>NLM ISO Abbreviation: </i>Bioresour Technol <i>Subsets: </i>MEDLINE
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  Data: <i>Original Publication</i>: Barking, Essex, England : New York, N.Y. : Elsevier Applied Science ; Elsevier Science Pub. Co., 1991-
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  Data: <searchLink fieldCode="MM" term="%22Bacillus+subtilis%22">Bacillus subtilis*</searchLink>/<searchLink fieldCode="MM" term="%22Bacillus+subtilis+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Bacillus+subtilis%22">Bacillus subtilis*</searchLink>/<searchLink fieldCode="MM" term="%22Bacillus+subtilis+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Bacillus+subtilis%22">Bacillus subtilis*</searchLink>/<searchLink fieldCode="MM" term="%22Bacillus+subtilis+enzymology%22">enzymology</searchLink> <br /><searchLink fieldCode="MM" term="%22Hexosyltransferases%22">Hexosyltransferases*</searchLink>/<searchLink fieldCode="MM" term="%22Hexosyltransferases+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Hexosyltransferases%22">Hexosyltransferases*</searchLink>/<searchLink fieldCode="MM" term="%22Hexosyltransferases+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Sucrose%22">Sucrose*</searchLink>/<searchLink fieldCode="MM" term="%22Sucrose+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Trisaccharides%22">Trisaccharides*</searchLink>/<searchLink fieldCode="MM" term="%22Trisaccharides+biosynthesis%22">biosynthesis</searchLink> <br /><searchLink fieldCode="MM" term="%22Lactose%22">Lactose*</searchLink>/<searchLink fieldCode="MM" term="%22Lactose+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Regulatory+Sequences%2C+Nucleic+Acid%22">Regulatory Sequences, Nucleic Acid*</searchLink>/<searchLink fieldCode="MM" term="%22Regulatory+Sequences%2C+Nucleic+Acid+genetics%22">genetics</searchLink><br /><searchLink fieldCode="MH" term="%22Hydrogen-Ion+Concentration%22">Hydrogen-Ion Concentration</searchLink> ; <searchLink fieldCode="MH" term="%22Gene+Expression+Regulation%2C+Bacterial%22">Gene Expression Regulation, Bacterial</searchLink> ; <searchLink fieldCode="MH" term="%22Fermentation%22">Fermentation</searchLink> ; <searchLink fieldCode="MH" term="%22Temperature%22">Temperature</searchLink> ; <searchLink fieldCode="MH" term="%22Promoter+Regions%2C+Genetic%22">Promoter Regions, Genetic</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Lactosucrose is a functional oligosaccharide with prebiotic properties and broad applications in the food and nutraceutical industries. In this study, a systematic strategy was developed to enhance lactosucrose production by optimizing levansucrase expression in Bacillus subtilis. Six levansucrases were screened, and levansucrase from Brenneria goodwinii (BgLS) exhibited the highest solubility and whole-cell catalytic activity. Whole-cell reaction conditions were optimized, identifying pH 6.0 and 50 °C as optimal. BgLS expression was further enhanced through translational regulation by engineering synthetic 5'-untranslated regions (5'-UTRs) and N-terminal coding sequences (NCSs), followed by combinatorial optimization. Subsequent transcriptional optimization using endogenous promoters further increased enzyme expression. The best engineered strain achieved a fermentation activity of 190.7 U/mL and produced lactosucrose with a 46.5% conversion rate from 200 g/L lactose and 300 g/L sucrose. This work demonstrates an efficient platform for high-level lactosucrose biosynthesis and highlights the effectiveness of multi-level gene expression regulation.<br /> (Copyright © 2026 Elsevier Ltd. All rights reserved.)
– Name: Abstract
  Label: Competing Interests
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  Data: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
– Name: SubjectMinor
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  Data: <i>Keywords: </i>Bacillus subtilis; Lactosucrose; Levansucrase; Translational and transcriptional regulation; Whole-cellbiocatalysis
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  Label: Substance Nomenclature
  Group: ID
  Data: EC 2.4.1.10 (levansucrase)<br />EC 2.4.1.- (Hexosyltransferases)<br />57-50-1 (Sucrose)<br />0 (Trisaccharides)<br />J2B2A4N98G (Lactose)
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  Data: 10.1016/j.biortech.2026.134954
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  Data: 42176820
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        Value: 10.1016/j.biortech.2026.134954
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      – Code: eng
        Text: English
    PhysicalDescription:
      Pagination:
        StartPage: 134954
    Subjects:
      – SubjectFull: Hydrogen-Ion Concentration
        Type: general
      – SubjectFull: Gene Expression Regulation, Bacterial
        Type: general
      – SubjectFull: Fermentation
        Type: general
      – SubjectFull: Temperature
        Type: general
      – SubjectFull: Promoter Regions, Genetic
        Type: general
      – SubjectFull: Bacillus subtilis genetics
        Type: general
      – SubjectFull: Bacillus subtilis metabolism
        Type: general
      – SubjectFull: Bacillus subtilis enzymology
        Type: general
      – SubjectFull: Hexosyltransferases metabolism
        Type: general
      – SubjectFull: Hexosyltransferases genetics
        Type: general
      – SubjectFull: Sucrose metabolism
        Type: general
      – SubjectFull: Trisaccharides biosynthesis
        Type: general
      – SubjectFull: Lactose metabolism
        Type: general
      – SubjectFull: Regulatory Sequences, Nucleic Acid genetics
        Type: general
    Titles:
      – TitleFull: Enhanced levansucrase expression in Bacillus subtilis via regulatory element optimization for high-yield lactosucrose production from sucrose and lactose.
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            – D: 01
              M: 09
              Text: 2026 Sep
              Type: published
              Y: 2026
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