Skip to main content

Advertisement

SpringerLink
Log in

Display of a sucrose isomerase on the cell surface of Yarrowia lipolytica for synthesis of isomaltulose from sugar cane by-products

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

Isomaltulose (α-d-glucopyranosyl-1,6-d-fructofuranose) is an important industrial and raw food material, which can be synthesised from the by-products of sugar cane processing through sucrose isomerization conversion. In this study, we constructed a surface display vector of sucrose isomerase from Pantoeadispersa (pSIase) by a glycosylphosphatidylinositol (GPI)-cell wall protein (CWP) anchor signal sequence and successfully displayed pSIase on the cell surface of Yarrowia lipolytica, thereby increasing the conversion efficiency of isomaltulose. The highest activity of the displayed pSIase reached 2910.3 U/g of cell dry weight. Compared with the free pSIase, the displayed enzyme showed good stability at a broad range of temperatures (20–45 °C). The half-life at 40 °C increased from 62 to 141 min and the deactivation constants (kd) reached 4.91 × 10−3 min−1. Using low-cost cane molasses as the substrate, the isomaltulose conversion rate remained at 85% even after 9 batches were processed, which is a highly desired outcome for industrial use.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Adachi N, Takahashi C, Ono-Murota N, Yamaguchi R, Tanaka T, Kondo A (2013) Direct L- lysine production from cellobiose by Corynebacterium glutamicum displaying beta-glucosidase on its cell surface. Appl Microbiol Biotechnol 97(16):7165–7172

    Article  CAS  Google Scholar 

  • Ahn SJ, Yoo JH, Lee HC, Kim SY, Noh BS, Kim JH, Lee JK (2003) Enhanced conversion of sucrose to isomaltulose by a mutant of Erwinia rhapontici. Biotechnol Lett 25:1179–1183

    Article  CAS  Google Scholar 

  • Aroonnual A, Nihira T, Seki T, Panbangred W (2007) Role of several key residues in the catalytic activity of sucrose isomerase from Klebsiella pneumoniae NK33-98-8. Enzyme Microb Technol 40:1221–1227

    Article  CAS  Google Scholar 

  • Cartarius R, Krause T, Vogel H (2001) Degradable surfactants via heterogeneously catalyzed reductive amination of isomaltulose-examination of catalyst deactivation in a continuous gradient free reaction vessel. Chem Eng Technol 24:55A–59A

    Google Scholar 

  • Cha J, Jung JH, Park SE, Cho MH, Seo DH, Ha SJ, Yoon JW, Lee OH, Kim YC, Park CS (2009) Molecular cloning and functional characterizationof a sucrose isomerase (isomaltulose synthase) gene from Enterobacter sp. FMB-1. J Appl Microbiol. 107:1119–1130

    Article  CAS  Google Scholar 

  • Cheetham PS (1984) The extraction and mechanism of novel isomaltulose synthesizing enzyme from Erwinia rhapontici. Biochem J 220(1):213–220

    Article  CAS  Google Scholar 

  • Cheetham PSJ, Imber CE, Isherwood J (1982) The formation of isomaltulose by immobilized Erwinia rhapontici. Nature 299:628–631

    Article  CAS  Google Scholar 

  • Chen YP, Hwang IE, Lin CJ, Wang HJ, Tseng CP (2012) Enhancing the stability of xylanase from Cellulomonas fimi by cell-surface display on Escherichia coli. J Appl Microbiol 112(3):455–463

    Article  CAS  Google Scholar 

  • Feldhaus M, Siegel R (2004) Flow cytometric screening of yeast surface display libraries. Methods Mol Biol 263:311–332

    CAS  PubMed  Google Scholar 

  • Fleddermann M, Rauh-Pfeiffer A, Demmelmair H, Holdt L, Teupser D, Koletzko B (2016) Effects of a Follow-on formula containing isomaltulose (PalatinoseTM) on metabolic response, acceptance, tolerance and safety ininfants: a randomized-controlled trial. PLoS One 11(3):e151614

  • Hamada S (2002) Role of sweeteners in the etiology and prevention of dentalcaries. Pure Appl Chem 74:1293–1300

    Article  CAS  Google Scholar 

  • Hiraishi T, Yamashita K, Sakono M, Nakanishi J, Tan LT, Sudesh K, Abe H, Maeda M (2012) Display of functionally active PHB depolymerase on Escherichia coli cell surface. Macromol Biosci 12(2):218–224

    Article  CAS  Google Scholar 

  • Jolivalt C, Madzak C, Brault A, Caminade E, Malosse C, Mougin C (2005) Expression of laccase lllb from the white-rot fungus Trametes versicolor in the yeast Yarrowia lipolytica for environmental applications. Appl Micro biol Biotechnol 66(4):450–456

    Article  CAS  Google Scholar 

  • Kawaguti HY, Sato HH (2007) Palatinose production by free and Ca-alginate gel immobilized cells of Erwinia sp. Biochem Eng J 36:202–208

    Article  CAS  Google Scholar 

  • Kuroda K, Matsui K, Higuchi S, Kotaka A, Sahara H, Hata Y, Ueda M (2009) Enhancement of display efficiency in yeast display system by vector engineering and gene disruption. Appl Microbiol Biotechnol 82(4):713–719

    Article  CAS  Google Scholar 

  • Lee HC, Kim JH, Kim SY, Lee JK (2008) Isomaltose production by modification of the fructose binding site on the basis of the predicted structure of sucrose isomerase from “Protaminobacter rubrum”. Appl Environ Microbiol. 74:5183–5194

    Article  CAS  Google Scholar 

  • Li S, Cai H, Qing Y, Ren B, Xu H, Zhu H, Yao J (2011) Cloning and characterization of a sucrose isomerase from Erwinia rhapontici NX-5 for isomaltulose hyperproduction. Appl Biochem Biotechnol. 163:52–63

    Article  CAS  Google Scholar 

  • Maeda RN, Barcelos CA, Santa Anna LM, Pereira N (2013) Cellulase production by Penicillium funiculosum and its application in the hydrolysis of sugar cane bagasse for second generation ethanol production by fed batch operation. J Biotechnol. 163:38–44

    Article  CAS  Google Scholar 

  • Ooshima T, Izumitani A, Minami T, Fujiwara T, Nakajima Y, Hamada S (1991) Trehalulose does not induce dental caries in rats infected with mutans Streptococci. Caries Res 25:277–282

    Article  CAS  Google Scholar 

  • Salvucci ME (2003) Distinct sucrose isomerases catalyze trehalulose synthesis in whiteflies, Bemisia argentifolii, and Erwinia rhapontici. Comp Biochem Physiol B 135:385–395

    Article  Google Scholar 

  • Schüürmann J, Quehl P, Festel G, Jose J (2014) Bacterial whole-cell biocatalysts by surface display of enzymes: toward industrial application. Appl Microbiol Biotechnol 98:8031–8046

    Article  Google Scholar 

  • Ueda M, Tanaka A (2000) Genetic immobilization of proteins on the yeast cell surface. Biotechnol Adv 18:121–140

    Article  CAS  Google Scholar 

  • Veronese T, Perlot P (1998) Proposition for the biochemical mechanism occurring inthesucrose isomerase active site. FEBS Lett 441(3):348–352

    Article  CAS  Google Scholar 

  • Veronese T, Perlot P (1999) Mechanism of sucrose conversion by the sucrose isomerase of Serratia plymuthica ATCC 15928. Enzyme Microb Technol 24:263–269

    Article  CAS  Google Scholar 

  • Watzlawick H, Mattes R (2009) Gene cloning, protein characterization and alteration of product selectivity for the trehalulose hydrolase and trehalulose synthase from “Pseudomonas mesoacidophila” MX-45. Appl Environ Microbiol 75:7026–7036

    Article  CAS  Google Scholar 

  • Wu L, Birch RG (2004) Characterization of Pantoea dispersa UQ68J: producer of a highly efficient sucrose isomerase for isomaltulose biosynthesis. J Appl Microbiol 97:93–103

    Article  Google Scholar 

  • Wu L, Birch RG (2005) Characterization of the highly efficient sucrose isomerase from UQ-68J and cloning of the sucrose isomerase gene. Appl Environ Microbiol 71(3):1581–1590

    Article  CAS  Google Scholar 

  • Yue LX, Chi ZM, Wang L, Liu J, Madzak C, Li J, Wang XH (2008) Construction of a new plasmid for surface display on cells of Yarrowia lipolytica. J Microbiol Methods 72:116–123

    Article  CAS  Google Scholar 

  • Yuzbasheva EY, Yuzbashev TV, Perkovskaya NI, Mostova EB, Vybornaya TV, Sukhozhenko AV, Toropygin IY, Sineoky SP (2015) Cell surface display of Yarrowia lipolytica lipase Lip2p using the cell wall protein YlPir1p, its characterization and application as a whole-cell biocatalyst. Appl Biochem Biotechnol 75:3888–3900

    Article  Google Scholar 

Download references

Acknowledgements

This study was supported by the grants from Central Public interset Scientific Institution Basal Research Funds, CAFS (no. 2017GH07) and 2017 green manufacturing projects of china (no. Z135060009002).

Author information

Authors and Affiliations

  1. Laboratory of Enzyme Engineering, Yellow Sea Fisheries Research Institute, Qingdao, 266071, People’s Republic of China

    Yuan Zheng, Zhipeng Wang, Xiaofeng Ji & Jun Sheng

Authors
  1. Yuan Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhipeng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaofeng Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jun Sheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun Sheng.

Ethics declarations

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zheng, Y., Wang, Z., Ji, X. et al. Display of a sucrose isomerase on the cell surface of Yarrowia lipolytica for synthesis of isomaltulose from sugar cane by-products. 3 Biotech 9, 179 (2019). https://doi.org/10.1007/s13205-019-1713-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-019-1713-9

Keywords

Search

Navigation