Overexpression and characterization of Aspergillus awamori wild-type and mutant glucoamylase secreted by the methylotrophic yeast Pichia pastoris: Comparison with wild-type recombinant glucoamylase produced using Saccharomyces cerevisiae and Aspergillus niger as hosts

Protein Expression and Purification

Volumn 9, Issue 2, 1997, Pages 159-170

  Fierobe, Henri Pierre ^a   Mirgorodskaya, Ekaterina ^b   Frandsen, Torben P ^a   Roepstorff, Peter ^b   Svensson, Birte ^a  

^a CARLSBERG LABORATORY   (Denmark)

^b UNIVERSITY OF SOUTHERN DENMARK   (Denmark)

Author keywords

[No Author keywords available]

Indexed keywords

ASPERGILLUS AWAMORI; ASPERGILLUS NIGER; PICHIA; PICHIA PASTORIS; SACCHAROMYCES CEREVISIAE;

EID: 0031105124     PISSN: 10465928     EISSN: None     Source Type: Journal    
DOI: 10.1006/prep.1996.0689     Document Type: Article

References (48)

1. Aleshin A. E., Golubev A., Firsov L. M., Honzatko R. B. Crystal structure of glucoamylase fromAspergillus awamoriX100. J. Biol. Chem. 267:1992;19291-19298.
2. Aleshin A. E., Firsov L. M., Honzatko R. B. Refined structure for the complex of acarbose with glucoamylase fromAspergillus awamoriX100. J. Biol. Chem. 269:1994;15631-15639.
3. Stoffer B., Aleshin A. E., Firsov L. M., Svensson B., Honzatko R. B. Refined structure for the complex of D-glucoAspergillus awamoriX100. FEBS Lett. 358:1995;57-61.
4. Olsen K., Svensson B., Christensen U. Stopped-flow fluorescence and steady-state kinetic studies of ligand-binding reactions of glucoamylase fromAspergillus niger. Eur. J. Biochem. 209:1992;777-784.
5. Olsen K., Christensen U., Sierks M. R., Svensson B. Reaction mechanism of Trp120 → Phe and wild-type glucoamylases fromAspergillus niger. Biochemistry. 32:1993;9686-9693.
6. B. Svensson, B. Stoffer, T. P. Frandsen, M. Søgaard, M. R. Sierks, K. W. Rodenburg, B. W. Sigurskjold, C. Dupont, 1994, Basic molecular features, mechanism and specificity of protein-carbohydrate interactions in amylolytic enzymes, Proceedings of Alfred Benzon Symposium 36, 202, 213, Munksgaard, Copenhagen.
7. Svensson B., Frandsen T. P., Matsui I., Juge N., Fierobe H.-P., Stoffer B., Rodenburg K. W. Mutational analysis of catalytic mechanism and specificity in amylolytic enzymes. Petersen S. B., Svensson B., Pedersen S. Carbohydrate Bioengineering. 1995;125-145 Elsevier Science, Amsterdam.
8. Boel E., Hjort I., Svensson B., Norris F., Norris K. E., Fiil N. P. Glucoamylases G1 and G2 fromAspergillus niger. EMBO J. 3:1984;1097-1102.
9. Nunberg J. H., Meade J. H., Cole G., Lawyer F. C., McCabe P., Schweickart V., Tal R., Wittman V. P., Flatgaard J. E., Innis M. Molecular cloning and characterization of the glucoamylase gene ofAspergillus awamori. Mol. Cell. Biol. 4:1984;2306-2315.
10. Svensson B., Larsen K., Svendsen I., Boel E. The complete amino acid sequence of the glycoprotein glucoamylase G1, fromAspergillus niger. Carlsberg Res. Commun. 48:1983;529-544.
11. Svensson B., Larsen K., Gunnarsson A. Characterization of a glucoamylase G2 fromAspergillus niger. Eur. J. Biochem. 154:1986;497-502.
12. Harris E. M. S., Aleshin A. E., Firsov L. M., Honzatko R. B. Refined structure for the complex of 1-deoxynojirimycin with glucoamylase fromAspergillus awamoriX100. Biochemistry. 32:1993;1618-1626.
13. Williamson G., Belshaw N. J., Williamson M. P. OAspergillus. Biochem. J. 282:1992;423-428.
14. Frandsen T. P., Dupont C., Lehmbeck J., Stoffer B., Sierks M. R., Honzatko R. B., Svensson B. Site directed mutagenesis of the catalytic base glutamic acid 400 in glucoamylase fromAspergillus niger. Biochemistry. 33:1994;13808-13816.
15. Clarke A. J., Svensson B. Identification of an essential tryptophanyl residue in the primary structure of glucoamylase G2 fromAspergillus niger. Carlsberg Res. Commun. 49:1984;559-566.
16. Sierks M. R., Ford C., Reilly P. J., Svensson B. Site-directed mutagenesis at the active site Trp120 ofAspergillus awamori. Prot. Eng. 2:1989;621-625.
17. Frandsen T. P., Christensen T., Stoffer B., Lehmbeck J., Dupont C., Honzatko R. B., Svensson B. Mutational analysis of the roles in catalysis and substrate recognition of arginines 54 and 305, aspartic acid 309, and tryptophan 317 located at subsites 1 and 2 in glucoamylase fromAspergillus niger. Biochemistry. 34:1995;10162-10169.
18. Berland C. R., Sigurskjold B. W., Stoffer B., Frandsen T. P., Svensson B. Thermodynamics of inhibitor binding to mutant forms of glucoamylase fromAspergillus niger. Biochemistry. 34:1995;10153-10161.
19. Chen H.-M., Ford C., Reilly P. J. Substitution of asparagine residues inAspergillus awamori. Biochem. J. 301:1994;275-281.
20. Fierobe H.-P., Stoffer B. B., Frandsen T. P., Svensson B. Mutational modulation of substrate bond-type specificity and thermostability of glucoamylase fromAspergillus awamori. Biochemistry. 35:1996;8696-8704.
21. Chen H.-M., Ford C., Reilly P. J. Identification and elimination by site-directed mutagenesis of thermolabile aspartyl bonds inAspergillus awamori. Prot. Eng. 8:1995;575-582.
22. Sierks M. R., Svensson B. Protein engineering of the relative specificity of glucoamylase fromAspergillus awamori. Prot. Eng. 7:1994;1479-1484.
23. Christensen T., Woeldike H., Boel E., Mortensen S. B., Hjortshoej K., Thim L., Hansen M. T. High level expression of recombinant genes inAspergillus oryzae. Bio/technology. 6:1988;1419-1422.
24. Svensson B., Pedersen T. G., Svendsen I., Sakai T., Ottesen M. Characterization of two forms of glucoamylase fromAspergillus niger. Carlsberg Res. Commun. 47:1982;55-69.
25. Romanos M. Advances in the use ofPichia pastoris. Curr. Opin. Biotechnol. 6:1995;527-533.
26. Cregg J. M., Tschopp J. F., Stillman C., Siegel R., Akong M., Craig W. S., Buckholz R. G., Madden K. R., Kellaris P. A., Davis G. R., Smiley B. L., Cruze J., Torregrossa R., Velicelebi G., Thill G. P. High level expression and efficient assembly of hepatitis B surface antigen in the methylotrophic yeast,Pichia pastoris. Bio/technology. 5:1987;479-485.
27. Tschopp J. F., Brust P., Cregg J. M., Stillman C. A., Gingeras T. R. Expression of thelacZPichia pastoris. Nucleic Acids Res. 15:1987;3859-3875.
28. Clare J. J., Rayment F. B., Ballantine S. P., Sreekhrishna K., Romanos M. A. High level expression of tetanus toxin fragment c inPichia pastoris. Bio/technology. 9:1991;455-460.
29. Barr K. A., Hopkins S. A., Sreekhrishna K. Protocol for efficient secretion of HSA developed fromPichia pastoris. Pharm. Eng. 12:1992;48-51.
30. Paifer E., Margolles E., Cremata J., Montesino R., Herrera L., Delgado J.-M. Efficient expression and secretion of recombinant alpha amylase inPichia pastoris. Yeast. 10:1994;1415-1459.
31. Søgaard M., Svensson B. Expression of cDNAs encoding barley α-amylase 1 and 2 in yeast and characterization of the secreted proteins. Gene (Amst.). 94:1990;173-179.
32. Innis M. A., Holland M. J., McCabe P. C., Cole G. E., Wittman V. P., Tal R., Watt W. K., Gelfand D. H., Holland J. P., Meade J. H. Expression, glycosylation, and secretion of anAspergillusSaccharomyces cerevisiae. Science. 228:1985;21-26.
33. Sambrook J., Fritsch E. F., Maniatis T. Molecular Cloning: A Laboratory Manual. 1989;Cold Spring Harbor Laboratory, Cold Spring Harbor.
34. Higuchi R., Krummel B., Saiki R. K. A general method forin vitro. Nucleic Acids Res. 16:1988;7351-7367.
35. Clarke A. J., Svensson B. The role of tryptophanyl residues in the function ofAspergillus niger. Carlsberg Res. Commun. 49:1984;111-122.
36. Stoffer B., Frandsen T. P., Busk P. K., Schneider P., Svendsen I., Svensson B. Production, purification and characterization of the catalytic domain of glucoamylase fromAspergillus niger. Biochem. J. 292:1993;197-202.
37. Karras M., Hillenkamp F. Laser desorption ionization of proteins with molecular masses exceeding 10000 Da. Anal. Chem. 60:1988;2299-2301.
38. Karras M., Ehring H., Nordhoff E., Stahl B., Strupat K., Hillenkamp F., Grehl M., Krebs B. Matrix-assisted laser desorption/ionization mass spectrometry with additives to 2,5-dihydroxybenzoic acid. Org. Mass. Spectrom. 28:1993;1476-1481.
39. Vestal M. L., Juhasz P., Martin S. A. Delayed extraction matrix-assisted laser desorption time-of-flight mass spectrometry. Rapid. Commun. Mass Spectrom. 9:1995;1044-1050.
40. Riddles P. W., Blakeley R. L., Zerner B. Reassessment of Ellman's reagent. Methods Enzymol. 91:1983;49-60.
41. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28:1956;350-356.
42. Palcic M., Skrydstrup T., Bock K., Le N., Lemieux R. U. Substrate recognition by amyloglucosidase: Evaluation of conformationally biased isomaltosides. Carbohydr. Res. 250:1993;87-92.
43. R. J. Leatherbarrow, 1987, Enzfitter, A Non-linear Regression Data Analysis Program for IBM PC, Elsevier Science, Amsterdam, The Netherlands.
44. Hiromi K., Nitta Y., Numata C., Ono S. Subsite affinities of glucoamylase: Examination of the validity of subsite theory. Biochim. Biophys. Acta. 302:1973;362-375.
45. Hiromi K., Ohnishi M., Tanaka A. Subsite structure and ligand binding mechanism of glucoamylase. Mol. Cell. Biochem. 51:1983;1-6.
46. Williamson G., Belshaw N. J., Noel T. R., Ring S. G., Williamson M. P. O. Eur. J. Biochem. 207:1992;661-670.
47. Neustroev K. N., Firsov L. M. Acid proteinase and the multiplicity of forms of glucoamylase fromAsp. awamori. Biokhimiya. 55:1990;776-785.
48. B. Stoffer, 1994, Structural and Functional Analysis of the Starch Hydrolyzing Enzyme Glucoamylase, Ph.D. Thesis, University of Copenhagen, Denmark.