Bio/Technology
9, 976 - 981 (1991)
doi:10.1038/nbt1091-976
Commercial Levels of Chymosin Production by AspergillusNigel S. Dunn-Coleman1, *, Peggy Bloebaum1, Randy M. Berka1, Elizabeth Bodie1, Nancy Robinson1, Gale Armstrong1, Michael Ward1, Melinda Przetak1, Gina L. Carter1, Regina LaCost1, Lori J. Wilson1, Katherine H. Kodama1, Enrique F. Baliu1, Ben Bower1, Michael Lamsa1
& Henry Heinsohn1
1Genencor International Inc., 180 Kimball Way, South San Francisco, CA 94080.
*Corresponding author We have increased the production of bovine chymosin in Aspergillus niger var. awamori to more than one gram per liter of secreted authentic enzyme by combining a mutagenesis protocol with a novel robotic screening program. Analysis of the superior chymosin producing strains indicated that they have enhanced capabilities to secrete extracellular proteins.
REFERENCES
- van Brunt, J. 1986. Fungi: The perfect hosts? Bio/Technology 4: 1057−1422. | ChemPort |
- Berka, R.M.andBarnett, C. 1989. The development of gene expression systems for filamentous fungi. Biotech. Adv. 7: 127−154. | Article | ISI | ChemPort |
- Christensen, T., Woeldike, H., Boel, E., Mortensen, S.B., Hjortshoej, K., Thim, L.andHansen, M.T. 1988. High level expression of recombinant genes in Aspergillus oryzae. Bio/Technology 6: 1419−1422. | ISI | ChemPort |
- Cullen, D., Gray, G.L., Wilson, L.J., Hayenga, K.J., Lamsa, M.H., Rey, M.W., Norton, S. and Berka, R.M. 1987. Controlled expression and secretion of bovine chymosin in Aspergillus nidulans. Bio/Technology 5: 369−376. | ISI | ChemPort |
- Gwynne, D.I., Buxton, F.P., Williams, S.A., Garven, S. and Davies, R.W. 1987. Genetically engineered secretion of active human interferon and a bacterial endoglucanase from Aspergillus nidulans. Bio/Technology 5: 713−719. | ISI | ChemPort |
- Upshall, A., Kumar, A.A., Bailey, M.C., Parker, M.D., Favreau, M.A., Lewison, K.P., Joeseph, M.L., Maraganore, M. and McKnight, G.L. 1987. Secretion of active human tissue plasminogen from the fungus Aspergillus nidulans. Bio/Technology 5: 1301−1304. | ISI | ChemPort |
- Harkki, J., Uusitalo, J., Bailey, M., Penttila, M. and Knowles, J.K.C. 1989. A novel fungal expression system: secretion of active calf chymosin from the filamentous fungus Trichoderma reesei. Bio/Technology 7: 169−174. | ISI | ChemPort |
- Foltmann, B. 1981. Gastric protease: Structure, function evolution and mechanism of action. Essays Biochem. 17: 53−84.
- Pedersen, V.B., Christensen, K.A. and Foltmann, B. 1979. Investigations on the activation of bovine prochymosin. Eur. J. Biol. Chem. 94: 573−580. | ChemPort |
- Ward, M. 1989 Production of calf chymosin by Aspergillus awamori, p. 288−294 In: Genetics and molecular Biology of Industrial Microorganisms. C. L. Hershberger, S.W. Queener and G. Hageman(Eds.). American Society for Microbiology, Washington, D.C.
- Ward, M. 1989. Heterologous gene expression in Aspergillus. In: Proceedings of the EMBO-Alko Workshop on Molecular Biology of Filamentous Fungi. H. Nevalainen and M. Penttila (Eds.). Foundation for Biotechnical and Industrial Fermentation Research 6: 119−128. | ChemPort |
- Ward, M., Wilson, L.J., Kodama, K.H., Rey, M.W. and Berka, R.M. 1990. Improved production of chymosin in Aspergillus by expression as a glucoamylase-chymosin fusion. Bio/Technology 8: 435−440. | PubMed | ISI | ChemPort |
- Lamsa, M. and Bloebaum, P. 1990. Mutation and screening to increase chymosin yield in a genetically engineered strain of Aspergillus awamori. J. Ind. Microbiol. 5: 229−238. | ISI | ChemPort |
- Berka, R.M., Ward, M., Wilson, L.J., Hayenga, K.J., Kodama, K.H., Carlomagno, L.P. and Thompson, S.A. 1990. Molecular cloning and deletion of the aspergillopepsin A gene from Aspergillus awamori. Gene. 86: 2153−162. | Article |
- Allen, K.E., McNally, M.T., Lowendorf, H.S., Slayman, C.W. and Free, S.J. 1989. Deoxyglucose-resistant mutants of Neurospora crassa: Isolation, mapping, and biochemical characterization. J. Bacteriol. 171: 53−58. | PubMed | ISI | ChemPort |
- Rowlands, R.T. and Turner, G. 1973. Nuclear and extranuclear inheritance of oligomycin resistance in Aspergillus nidulans. Mol. Gen. Genet. 126: 201−216. | PubMed | ISI | ChemPort |
- van Hartingsveldt, W., Mattern, I.E., van Zieijl, C.M.J., Pouwels, P.H. and van den Hondel, C.A.M.J.J. 1987. Development of a homologous transformation system for Aspergillus niger based on the pyrG gene. Mol. Gen. Genet. 206: 71−75. | PubMed | ChemPort |
- Campbell, E.I., Unkles, S.E., Macro, J.A., Van den Hondel, C., Contreras, R. and Kinghorn, J.R. 1989. Improved transformation efficiency of Aspergillus niger using the homologous niaD gene for nitrate reductase. Curr. Genet. 16: 53−65. | PubMed | ISI | ChemPort |
- Kelly, J.M. and Hynes, M. 1985. Transformation of Aspergillus niger by the amdS gene from Aspergillus nidulans. EMBO J. 4: 475−479. | PubMed | ISI | ChemPort |
- Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, N.Y.
- Carlton, B.C. and Brown, B.J. 1981. Gene mutation, 222−232. In: Manual of Methods for General Bacteriology. P. Gerhardt(Ed.). American Society for Microbiology. Washington, D.C.
- Ohnishi, S.T. and Barr, J.K. 1978. A simplified method of quantitating proteins using the biuret and phenol reagents. Anal. Biochem. 86: 193−197. | PubMed | ISI | ChemPort |
- Laemmili, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680−685. | PubMed |
- Sleep, D., Belfield, G.P., Ballance, D.J., Steven, J., Jones, S., Evans, L.R., Moir, P.D. and Goodey, A.R. 1991. Saccharomyces cerevisiae strains that overexpress heterologous proteins. Bio/Technology 9: 183−186. | PubMed | ISI | ChemPort |
- van den Berg, J.A., van der Laken, K.J., van Ooyen, J.J., Renniers, T.C.H.M., Rietveld, K. and Schaap, A., Brake, A.J., Bishop, R.J., Schultz, K., Moyer, D., Richman, M. and FShuster, J.R. 1990. Kluyveromyces as a host for heterologous gene expression: Expression and secretion of prochymosin. Bio/Technology 8: 135−139. | PubMed | ChemPort |
|
