Advertisement

3 Biotech

, 9:179 | Cite as

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

  • Yuan Zheng
  • Zhipeng Wang
  • Xiaofeng Ji
  • Jun ShengEmail author
Original Article
  • 17 Downloads

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.

Keywords

Sugar cane Isomaltulose Sucrose isomerase Cell surface display 

Notes

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).

Compliance with ethical standards

Conflict of interest

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

References

  1. 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):7165–7172CrossRefGoogle Scholar
  2. 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 25:1179–1183CrossRefGoogle Scholar
  3. 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 40:1221–1227CrossRefGoogle Scholar
  4. 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–59AGoogle Scholar
  5. 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. 107:1119–1130CrossRefGoogle Scholar
  6. Cheetham PS (1984) The extraction and mechanism of novel isomaltulose synthesizing enzyme from Erwinia rhapontici. Biochem J 220(1):213–220CrossRefGoogle Scholar
  7. Cheetham PSJ, Imber CE, Isherwood J (1982) The formation of isomaltulose by immobilized Erwinia rhapontici. Nature 299:628–631CrossRefGoogle Scholar
  8. 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):455–463CrossRefGoogle Scholar
  9. Feldhaus M, Siegel R (2004) Flow cytometric screening of yeast surface display libraries. Methods Mol Biol 263:311–332PubMedGoogle Scholar
  10. 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):e151614CrossRefGoogle Scholar
  11. Hamada S (2002) Role of sweeteners in the etiology and prevention of dentalcaries. Pure Appl Chem 74:1293–1300CrossRefGoogle Scholar
  12. 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):218–224CrossRefGoogle Scholar
  13. 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):450–456CrossRefGoogle Scholar
  14. Kawaguti HY, Sato HH (2007) Palatinose production by free and Ca-alginate gel immobilized cells of Erwinia sp. Biochem Eng J 36:202–208CrossRefGoogle Scholar
  15. 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):713–719CrossRefGoogle Scholar
  16. 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. 74:5183–5194CrossRefGoogle Scholar
  17. 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. 163:52–63CrossRefGoogle Scholar
  18. 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. 163:38–44CrossRefGoogle Scholar
  19. 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 25:277–282CrossRefGoogle Scholar
  20. Salvucci ME (2003) Distinct sucrose isomerases catalyze trehalulose synthesis in whiteflies, Bemisia argentifolii, and Erwinia rhapontici. Comp Biochem Physiol B 135:385–395CrossRefGoogle Scholar
  21. Schüürmann J, Quehl P, Festel G, Jose J (2014) Bacterial whole-cell biocatalysts by surface display of enzymes: toward industrial application. Appl Microbiol Biotechnol 98:8031–8046CrossRefGoogle Scholar
  22. Ueda M, Tanaka A (2000) Genetic immobilization of proteins on the yeast cell surface. Biotechnol Adv 18:121–140CrossRefGoogle Scholar
  23. Veronese T, Perlot P (1998) Proposition for the biochemical mechanism occurring inthesucrose isomerase active site. FEBS Lett 441(3):348–352CrossRefGoogle Scholar
  24. Veronese T, Perlot P (1999) Mechanism of sucrose conversion by the sucrose isomerase of Serratia plymuthica ATCC 15928. Enzyme Microb Technol 24:263–269CrossRefGoogle Scholar
  25. 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 75:7026–7036CrossRefGoogle Scholar
  26. Wu L, Birch RG (2004) Characterization of Pantoea dispersa UQ68J: producer of a highly efficient sucrose isomerase for isomaltulose biosynthesis. J Appl Microbiol 97:93–103CrossRefGoogle Scholar
  27. 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):1581–1590CrossRefGoogle Scholar
  28. 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 72:116–123CrossRefGoogle Scholar
  29. 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 75:3888–3900CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Laboratory of Enzyme EngineeringYellow Sea Fisheries Research InstituteQingdaoPeople’s Republic of China

Personalised recommendations