Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The Arxula adeninivoransATAL gene encoding transaldolase-gene characterization and biotechnological exploitation

  • 94 Accesses

  • 10 Citations

Abstract

The yeast Arxulaadeninivorans provides an attractive expression platform and can be exploited as gene source for biotechnologically interesting proteins. In the following study, a striking example for the combination of both aspects is presented. The transaldolase-encoding A. adeninivorans ATAL gene, including its promoter and terminator elements, was isolated and characterized. The gene includes a coding sequence of 963 bp encoding a putative 321 amino acid protein of 35.0 kDa. The enzyme characteristics analyzed from isolates of native strains and recombinant strains overexpressing the ATAL gene revealed a molecular mass of ca. 140 kDa corresponding to a tetrameric structure, a pH optimum of ca. 5.5, and a temperature optimum of 20°C. The preferred substrates for the enzyme include d-erythrose-4-phosphate and d-fructose-6-phosphate, whereas d-glyceraldehyde is not converted. The ATAL expression level under salt-free conditions was observed to increase in media supplemented with 5% NaCl rendering the ATAL promoter attractive for moderate heterologous gene expression under high-salt conditions. Its suitability was assessed for the expression of a human serum albumin (HSA) reporter gene.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Böer E, Wartmann T, Dlubatz K, Gellissen G, Kunze G (2004a) Characterization of the Arxula adeninivorans AHOG1 gene and the encoded mitogen-activated protein kinase. Curr Genet 46:269–276

  2. Böer E, Wartmann T, Luther B, Manteuffel R, Bode R, Gellissen G, Kunze G (2004b) Characterization of the AINV gene and the encoded invertase from the dimorphic yeast Arxula adeninivorans. Antonie van Leeuwenhoek 86:121–134

  3. Böer E, Gellissen G, Kunze G (2005a) Arxula adeninivorans. In: Gellissen G (ed) Production of recombinant proteins. Wiley, Weinheim, pp 89–110

  4. Böer E, Mock HP, Bode R, Gellissen G, Kunze G (2005b) An extracellular lipase from the dimorphic yeast Arxula adeninivorans: molecular cloning of the ALIP1 gene and characterization of the purified recombinant enzyme. Yeast 22:523–535

  5. Böer E, Wartmann T, Schmidt S, Bode R, Gellissen G, Kunze G (2005c) Characterization of the AXDH gene and the encoded xylitol dehydrogenase from the dimorphic yeast Arxula adeninivorans. Antonie van Leeuwenhoek 87:233–243

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  7. Bui DM, Kunze I, Förster S, Wartmann T, Horstmann C, Manteuffel R, Kunze G (1996) Cloning and expression of an Arxula adeninivorans glucoamylase gene in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 44:610–619

  8. Dohmen RJ, Strasser AM, Höner CB, Hollenberg CP (1991) An efficient transformation procedure enabling long-term storage of competent cells of various yeast genera. Yeast 7:691–692

  9. Gellissen G (2005) Production of recombinant proteins. Wiley, Weinheim

  10. Gellissen G, Kunze G, Gaillardin C, Cregg JM, Berardi E, Veenhuis M, van der Klei I (2005) New yeast expression platforms based on methylotrophic Hansenula polymorpha and Pichia pastoris and on dimorphic Arxula adeninivorans and Yarrowia lipolytica—a comparison. FEMS Yeast Res 5:1079–1096

  11. Gienow U, Kunze G, Schauer F, Bode R, Hofemeister J (1990) The yeast genus Trichosporon spec. LS3; molecular characterization of genomic complexity. Zentralbl Mikrobiol 145:3–12

  12. Horecker BL, Smyrniotis PZ (1955) Purification and properties of yeast transaldolase. J Biol Chem 212:811–825

  13. Horecker BL, Tsolas O (1990) Purification and crystallization of transaldolase from Candida utilis. Methods Enzymol 182:788–793

  14. Johansson B, Hahn-Hägerdal B (2002) The non-oxidative pentose phosphate pathway controls the fermentation rate of xylulose but not of xylose in Saccharomyces cerevisiae TMB3001. FEMS Yeast Res 2:277–282

  15. Kaur P, Lingner A, Singh B, Böer E, Polajeva J, Steinborn G, Bode R, Gellissen G, Satyanarayana T, Kunze G (2007) APHO1 from the yeast Arxula adeninivorans encodes an acid phosphatase of broad substrate specificity. Antonie van Leeuwenhoek 91:45–55

  16. Kobori Y, Myles DC, Whitesides GM (1992) Substrate specificity and carbohydrate synthesis using transketolase. J Org Chem 57:5899–5907

  17. Lachaise F, Martin G, Drougard C, Perl A, Vuillaume M, Wegnez M, Sarasin A, Daya-Grosjean L (2001) Relationship between posttranslational modification of transaldolase and catalase deficiency in UV-sensitive repair-deficient Xeroderma pigmentosum fibroblsts and SV40-transformed human cells. Free Radic Biol Med 15:1365–1373

  18. Middelhoven JW, Hoogkamer-Te Niet MC, Kreger van Rij NJW (1984) Trichosporon adeninovorans sp. nov., a yeast species utilizing adenine, xanthine, uric acid, putrescine and primary n-alkylamines as the sole source of carbon, nitrogen and energy. Antonie van Leeuwenhoek 50:369–387

  19. Middelhoven WJ, de Jonge IM, Winter M (1991) Arxula adeninivorans, a yeast assimilating many nitrogenous and aromatic compounds. Antonie van Leeuwenhoek 60:129–137

  20. Middelhoven WJ, Coenen A, Kraakman B, Gelpke MDS (1992) Degradation of some phenols and hydroxybenzoates by the imperfect ascomycetous yeasts Candida parapsilosis and Arxula adeninivorans: evidence for an operative gentisate pathway. Antonie van Leeuwenhoek 62:181–187

  21. Rösel H, Kunze G (1995) Cloning and characterization of a TEF gene for elongation factor 1α from the yeast Arxula adeninivorans. Curr Genet 28:360–366

  22. Rösel H, Kunze G (1998) Integrative transformation of the dimorphic yeast Arxula adeninivorans LS3 based on hygromycin B resistance. Curr Genet 33:157–163

  23. Samsonova IA, Kunze G, Bode R, Böttcher F (1996) A set of genetic markers for the chromosomes of the imperfect yeast Arxula adeninivorans. Yeast 12:1209–1217

  24. Sprenger GA, Schörken U, Sprenger G, Sahm H (1995) Transaldolase B of Escherichia coli K-12: cloning of its gene, talB, and characterization of the enzyme from recombinant strains. J Bacteriol 177:5930–5936

  25. Steinborn G, Gellissen G, Kunze G (2005) Assessment of Hansenula polymorpha and Arxula adeninivorans-derived rDNA-targeting elements for the design of Arxula adeninivorans expression vectors. FEMS Yeast Res 5:1047–1054

  26. Steinborn G, Wartmann T, Gellissen G, Kunze G (2007) Construction of an Arxula adeninivorans host-vector system based on trp1 complementation. J Biotechnol 127:392–401

  27. Terentiev Y, Gellissen G, Kunze G (2003) Arxula adeninivorans—a non-conventional dimorphic yeast of great biotechnological potential. Recent Res Devel Applied Microbiol Biotech 1:135–145

  28. Terentiev Y, Pico AH, Böer E, Wartmann T, Klabunde J, Breuer U, Babel W, Suckow M, Gellissen G, Kunze G (2004a) A wide-range integrative yeast expression vector system based on Arxula adeninivorans-derived elements. J Ind Microbiol Biotech 31:223–228

  29. Terentiev Y, Breuer U, Babel W, Kunze G (2004b) Non-conventional yeasts as producers of polyhydroxyalkanoates—genetic engineering of Arxula adeninivorans. Appl Microbiol Biotechnol 64:376–381

  30. Tsolas O, Horecker BL (1972) Transaldolase. In: Boyer PD (ed) The enzymes, 3rd edn, vol 7. Academic, New York, pp 259–280

  31. Van der Walt JP, Smith MT, Yamada Y (1990) Arxula gen. nov. (Candidaceae), a new anamorphic, arthroconidial yeast genus. Antonie van Leeuwenhoek 57:59–61

  32. Wahlbom CF, Otero RRC, van Zyl WH, Hahn-Hägerdal B, Jönsson LJ (2003) Molecular analysis of a Saccharomyces cerevisiae mutant with improved ability to utilize xylose shows enhanced expression of proteins involved in transport, initial xylose metabolism, and the pentose phosphate pathway. Appl Environ Microbiol 69:740–746

  33. Wartmann T, Krüger A, Adler K, Bui MD, Kunze I, Kunze G (1995) Temperature dependent dimorphism of the yeast Arxula adeninivorans LS3. Antonie van Leeuwenhoek 68:215–223

  34. Wartmann T, Böer E, Huarto-Pico A, Sieber H, Bartelsen O, Gellissen G, Kunze G (2002) High-level production and secretion of recombinant proteins by the dimorphic yeast Arxula adeninivorans. FEMS Yeast Res 2:363–369

  35. Wartmann T, Stoltenburg R, Böer E, Sieber H, Bartelsen O, Gellissen G, Kunze G (2003a) The ALEU2 gene—a new component for an Arxula adeninivorans-based expression platform. FEMS Yeast Res 3:223–232

  36. Wartmann T, Bellebna C, Böer E, Gellissen G, Kunze G (2003b) The constitutive AHSB4 promoter—a novel component of the Arxula adeninivorans-based expression platform. Appl Microbiol Biotechnol 62:528–535

  37. Wolf K (1996) Nonconventional yeasts in biotechnology. Springer, Berlin Heidelberg New York

  38. Yang XX, Wartmann T, Stoltenburg R, Kunze G (2000) Halotolerance of the yeast Arxula adeninivorans LS3. Antonie van Leeuwenhoek 77:303–311

Download references

Acknowledgements

We are grateful to Dr. I. Kunze for the helpful discussions and critical reading of the manuscript. We also thank I. Schmeling and H. Bohlmann for their excellent technical assistance. The research work was supported by grants from the “Ministry of Science and Research,” Sachsen/Anhalt (grant nos. 2067A/0025, 2463A/0086G), “Ministry of Economics,” Nordrhein-Westfalen (TPW-9910v08), and by Funds of Chemical Industry (GK).

Author information

Correspondence to Gotthard Kunze.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Fiki, A.E., Metabteb, G.E., Bellebna, C. et al. The Arxula adeninivoransATAL gene encoding transaldolase-gene characterization and biotechnological exploitation. Appl Microbiol Biotechnol 74, 1292–1299 (2007). https://doi.org/10.1007/s00253-006-0785-8

Download citation

Keywords

  • Arxula adeninivorans
  • ATAL
  • Heterologous gene expression
  • Transaldolase
  • Yeast