Abstract
The glucoamylase gene of the yeast Arxula adeninivorans was expressed in Kluyveromyces lactis by using the GAP promoter from Saccharomyces cerevisiae and a multicopy plasmid vector. The transformants secreted 90.1% of the synthesized glucoamylase into the culture medium. The secreted glucoamylase activities are about 20 times higher in comparison to those of Saccharomyces cerevisiae transformants using the same promoter. Secreted glucoamylase possesses identical N-terminal amino acid sequences to those secreted by A. adeninivorans showing that cleavage of the N-terminal signal peptide takes place at the same site. Biochemical characteristics of glucoamylase expressed by K. lactis and A. adeninivorans are very similar.
Similar content being viewed by others
References
Baldari C, Murray JAH, Ghiara P, Cesareni G, Galeotti CL (1987) A novel leader peptide which allows efficient secretion of a fragment of human interleukin 1β in Saccharomyces cerevisiae. EMBO J 6: 229–234
Bergkamp RJM, Kool IM, Geerse RH, Planta RJ (1992) Multiplecopy integration of the α-galactosidase gene from Cyamopsis tetragonoloba into the ribosomal DNA of Kluyveromyces lactis. Curr Genet 21: 365–370
Bianchi MM (1992) Site-specific recombination of the circular 2μm-like plasmid pKD1 requires integrity of the recombinase A and of the partitioning genes B and C. J Bacteriol 174: 6703–6706
Bianchi MM, Santarelli R, Frontali L (1991) Plasmid functions involved in the stable propagation of pKD1 circular plasmid in Kluyveromyces lactis. Curr Genet 19: 155–161
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7: 1513 1523
Bradford MH (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
Breunig KD, Kuger P (1987) Functional homology between the yeast regulatory proteins GAL4 and LAC9: LAC9-mediated transcriptional activation in Kluyveromyces lactis involves protein binding to a regulatory sequence homologous to the GAL4 protein-binding site. Mol Cell Biol 7: 4400–4406
Bui MD. Kunze I. Förster S. Wartmann T, Horstmann C, Manteuffel R, Kunze G (1995) Cloning and expression of an Arxula adeninivorans glucoamylase gene in S. cerevisiae. Appl Microbiol Biotechnol (in press)
Büttner R, Bode R, Birnbaum D (1987) Purification and characterization of the extracellular glucoamylase from the yeast Trichosporon adeninovorans. J Basic Microbiol 27: 299–308
Chen XJ, Saliola M, Falcone C, Bianchi MM, Fukuhara H (1986) Sequence organization of the circular plasmid pKD1 from the yeast Kluyveromyces drosophilarum. Nucleic Acids Res 14: 4471–4481
Davis E, Larkins EA, Knight RH (1972) Polysomes from peas. An improved method for their isolation in the absence of ribonuclease inhibitors. Plant Physiol 50: 581–584
Dohmen RJ, Strasser WM, Hörner CB, Hollenberg CP (1991) An efficient transformation procedure enabling long-term storage of competent cells of various yeast genera. Yeast 7: 691–692
Fleer R (1992) Engineering yeast for high level expression. Curr Opin in Biotechnol 3: 486–496
Fleer R. Yeh P, Amellal N, Fournier A. Bacchetta F. Baduel P, Jung G, L’Hote H, Becquart J, Fukuhara H, Mayaux JF (1991) Stable multicopy vectors for high-level secretion of recombinant human serum albumin in Kluyveromyces yeast. Biotechnology 9: 968–975
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
Hanahan D (1983) Studies on transformation of E. coli with plasmids. J Mol Biol 166: 557–580
Middelhoven WJ, 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 of carbon, nitrogen and energy. Antonie Van Leeuwenhoek 50: 369–378
Middelhoven WJ, Jong IM de, Winter M de (1991) Arxula adeninivorans, a yeast assimilating many nitrogenous and aromatic compounds. Antonie Van Leeuwenhoek 60: 129–137
Romanos MA, Scorer CA, Clare JJ (1992) Foreign gene expression in yeast: a review. Yeast 8: 423–488
Rose MD, Winston F, Hieter P (1990) Methods in yeast genetics. A laboratory course manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning — a laboratory manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, NY
Sherman F, Fink GR, Hicks JB (1986) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Stark MJR, Milner JS (1989) Cloning and analysis of the Kluyveromyces TRP1 gene: a chromosomal locus flanked by genes encoding inorganic pyrophosphatase and histone H3. Yeast 5: 35–50
Stoltenburg R, Wartmann T, Kunze I, Kunze G (1995) Reliable method to prepare RNA from free and membrane-bound polysomes from different yeast species. Biotechnology 18: 564–568
Van den Berg JA, Van der Lanken KJ, Van Ooyen AJJ, Renniers CHM, Rietveld K, Schaap A, Brake AJ, Bishop RJ, Schultz K, Moyer D, Richman M, Shuster JR (1990) Kluyveromyces as a host for heterologous gene expression: expression and secretion of prochymosin. Bio-Techn 8: 135–139
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
Wartmann T, Kunze I, Bui MD, Manteuffel R, Kunze G (1995) Comparative biochemical and immunological studies of glucoamylase producing Arxula adeninivorans yeast strains. Microbiol Res 150: 113–120
Yeh P, Landais D, Lemaitre M, Maury I, Crenne JY, Becquart J, Murry-Brelier A, Boucher F, Montay G, Fleer R (1992) Design of yeast-secreted albumin derivatives for human therapy — biological and antiviral properties of a serumalbumin-CD4 genetic conjugate. Proc Natl Acad Sci USA 89: 1904–1908
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bui, D.M., Kunze, I., Horstmann, C. et al. Expression of the Arxula adeninivorans glucoamylase gene in Kluyveromyces lactis . Appl Microbiol Biotechnol 45, 102–106 (1996). https://doi.org/10.1007/s002530050655
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s002530050655