Summary
An enzyme with α-galactosidase activity and an apparent molecular weight of 82 kDa was purified from culture medium of Aspergillus niger. The N-terminal amino acid sequence of the purified protein shows similarity to the N-terminal amino acid sequence of α-galactosidases from several other organisms. Oligonucleotides, based on the N-terminal amino acid sequence, were used as probes to clone the corresponding gene from a λ EMBL3 gene library of A. niger. The cloned gene (aglA) was shown to be functional by demonstrating that the 82 kDa α-galactosidase is absent from a strain with a disruption of the agIA gene, and is over-produced in strains containing multiple copies of the aglA gene. Enzyme activity assays revealed that the 82 kDa α-galactosidase A represents a minor extracellular α-galactosidase activity in A. niger.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bahl OP, Agrawal KML (1972) Degradation of complex carbohydrates. α-galactosidase, β-galactosidase, and β-N-acetylglucosaminidase from A. niger. Methods Enzymol 28:728–734
Barbesgaard P (1977) Industrial enzymes produced by members of the genus Aspergillus. In: Smith JE, Pateman JA (eds) Genetics and physiology of Aspergillus. Br Myc Soc Symp Series no. 1. Academic Press, London, pp 391–404
Bennett JW (1985) Molds, manufacturing and molecular genetics. In: Timberlake WE (ed) Molecular genetics of filamentous fungi. Alan R Liss, New York, pp 345–366
Bennett JW, Lasure LL (1991) Growth media. In: Bennett JW, Lasure LL (eds) More gene manipulation in fungi. Academic Press, San Diego, California, pp 441–458
Bos CJ (1986) Induced mutation and somatic recombination as tools for genetic analysis and breeding of imperfect fungi. Thesis, Agricultural University, Wageningen, The Netherlands
Chen EY, Seeburg PH (1985) Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA 4:165–170
Civas A, Eberhard R, Le Dizet P, Petek F (1984a) Glycosidases induced in Aspergillus tamarii. Mycelial α-d-galactosidases. Biochem J 219:849–855
Civas A, Eberhard R, Le Dizet P, Petek F (1984b) Glycosidases induced in Aspergillus tamarii. Secreted α-d-galactosidase and β-d-mannanase. Biochem J 219:857–863
Dey PM, Pridham JB (1972) Biochemistry of α-galactosidases. Adv Enzymol 36:91–130
Frischauf AM, Lehrach H, Poustka A, Murray N (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842
Gurr SJ, Unkles SE, Kinghorn JR (1988) The structure and organization of nuclear genes of filamentous fungi. In: Kinghorn JR (ed) Gene structure in eukaryotic microbes SGM special publication, vol 23. IRL Press, Oxford, pp 93–139
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
van Hartingsveldt W, Mattern IE, van Zeijl CMJ, Pouwels PH, van den Hondel CAMJJ (1987) Development of a homologous transformation system for Aspergillus niger based on the pyrG gene. Mol Gen Genet 206:71–75
von Heijne G (1983) Patterns of aminoacids near signal sequence cleavage sites. Eur J Biochem 133:17–21
von Heijne G, Abrahmsen L (1989) Species-specific variation in signal peptide design. FEBS Lett 244:439–446
van den Hondel CAME, Punt PJ, Gorcom RMF (1991) Heterologous gene expression in filamentous fungi. In: Benett JW, Lasure LL (eds) More gene manipulation in fungi. Academic Press, San Diego, California, pp 396–428
Hynes MJ, Corrick CM, King JA (1983) Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol Cell Biol 3:1430–1439
Kornreich R, Desnick RJ, Bishop DF (1989) Nucleotide sequence of the human α-galactosidase A gene. Nucleic Acids Res 17:3301–3302
Overbeeke N, Fellinger AM, Toonen MY, van Wassenaar D, Verrips CT (1989) Cloning and nucleotide sequence of the α-galactosidase cDNA from Cyamopsis tetragonoloba (guar). Plant Mol Biol 13:541–550
Penttila M, Lethovaara P, Nevalainen H, Bhikhabbai R, Knowles J (1986) Homology between cellulase genes from Trichoderma reesei: complete nucleotide sequence of the endoglucanase gene I. Gene 45:253–263
Punt PJ, Oliver RP, Dingemanse MA, Pouwels P, van den Hondel CAMJJ (1987) Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli. Gene 56:117–124
Punt PJ, Dingemanse MA, Kuyvenhoven A, Soede RDM, Pouwels PH, van den Hondel CAMJJ (1990) Functional elements in the promoter region of the Aspergillus nidulans gpdA gene encoding glyceraldehyde-3-phosphate dehydrogenase. Gene 93:101–109
de Ruiter-Jacobs YMJT, Broekhuijsen M, Unkles SE, Campbell El, Kinghorn JR, Contreras R, Pouwels PH, van den Hondel CAMJJ (1989) A gene transfer system based on the homologous pyrG gene and efficient expression of bacterial genes in Aspergillus oryzae. Curr Genet 16:159–163
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual, 2nd. edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, USA
Sumner-Smith M, Bozzato RP, Skipper N, Wayne Davies R, Hopper JE (1985) Analysis of the inducible MEL1 gene of Saccharomyces carlsbergensis and its secreted product, alpha-galactosidase (melibiase). Gene 36:333–340
Svensson B, Larson K, Svendsen I, Boel E (1983) The complete amino acid sequence of the glycoprotein, glucoamylase GI, from Aspergillus niger. Carlsberg Res Commun 48:529–544
Turner G, Brown J, Kerry-Williams S, Bailey AM, Ward M, Punt PJ, van den Hondel CAMJJ (1989) Analysis of the oliC promoter of Aspergillus nidulans. In: Nevalainen H, Penttila M (eds) Proceeding of the EMBO-Alko workshop on Molecular Biology of Filamentous Fungi. Foundation for Biotechnical and Industrial Fermentation Research, Helsinki, pp 101–109
Wernars K (1986) DNA-mediated transformation of the filamentous fungus Aspergillus nidulans. Thesis, Agricultural University, Wageningen, The Netherlands
Yanish-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Author information
Authors and Affiliations
Additional information
Communicated by W. Gajewski
Rights and permissions
About this article
Cite this article
den Herder, I.F., Mateo Rosell, A.M., van Zuilen, C.M. et al. Cloning and expression of a member of the Aspergillus niger gene family encoding α-galactosidase. Molec. Gen. Genet. 233, 404–410 (1992). https://doi.org/10.1007/BF00265437
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00265437