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. 1977 Jan;33(1):59–64. doi: 10.1128/aem.33.1.59-64.1977

Production and Purification of Thermostable Amylase and Protease of Thermomonospora viridis

Mary E Upton 1, William M Fogarty 1
PMCID: PMC170574  PMID: 16345190

Abstract

Maximum yields of amylase were produced by the thermophilic actinomycete Thermomonospora viridis in a modified Simpson and McCoy medium containing 1.5% corn starch and 0.5% mycological peptone with an initial pH 7.0. Best yields of amylase were obtained after incubation for 48 h, when the pH of the medium had risen to 8.2. Amylase was purified 313-fold by precipitation with n-propyl alcohol, dialysis against tap water, adsorption on Ca3(PO4)2, and fractionation on Sephadex G-100. Protease was produced in nutrient broth containing 0.5% starch and 1.0% corn steep liquor and at an initial pH 7.0. Maximum yields of protease were produced after 42 h. The protease was purified 54-fold by precipitation with n-propyl alcohol, dialysis against tap water, adsorption on Ca3(PO4)2, and fractionation on Sephadex G-200.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. CAMPBELL L. L., Jr Purification and properties of an alpha-amylase from facultative thermophilic bacteria. Arch Biochem Biophys. 1955 Jan;54(1):154–161. doi: 10.1016/0003-9861(55)90018-7. [DOI] [PubMed] [Google Scholar]
  2. COLEMAN G., ELLIOTT W. H. Studies on alpha-amylase formation by Bacillus subtilis. Biochem J. 1962 May;83:256–263. doi: 10.1042/bj0830256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. HARTMAN P. A., WELLERSON R., Jr, TETRAULT P. A. Bacillus stearothermophilus. I. Thermal and pH stability of the amylase. Appl Microbiol. 1955 Jan;3(1):7–10. doi: 10.1128/am.3.1.7-10.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. KUSTER E., LOCCI R. Studies on peat and peat microorganism. I. Taxonomic studies on thermophilic Actinomycetes isolated from peat. Arch Mikrobiol. 1963;45:188–197. doi: 10.1007/BF00408439. [DOI] [PubMed] [Google Scholar]
  5. Kuo M. J., Hartman P. A. Isolation of amylolytic strains of Thermoactinomyces vulgaris and production of thermophilic actinomycete amylases. J Bacteriol. 1966 Sep;92(3):723–726. doi: 10.1128/jb.92.3.723-726.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. MASSEY V. The relation of diaphorase and cytochrome reductase. Biochim Biophys Acta. 1960 Jan 15;37:310–314. doi: 10.1016/0006-3002(60)90238-9. [DOI] [PubMed] [Google Scholar]
  8. Mizusawa K., Ichishima E., Yoshida F. Production of thermostable alkaline proteases by thermophilic Streptomyces. Appl Microbiol. 1969 Mar;17(3):366–371. doi: 10.1128/am.17.3.366-371.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Morihara K., Tsuzuki H., Oka T. Comparison of the specificities of various neutral proteinases from microorganisms. Arch Biochem Biophys. 1968 Mar 11;123(3):572–588. doi: 10.1016/0003-9861(68)90179-3. [DOI] [PubMed] [Google Scholar]
  10. Ong P. S., Gaucher G. M. Protease production by thermophilic fungi. Can J Microbiol. 1973 Jan;19(1):129–133. doi: 10.1139/m73-019. [DOI] [PubMed] [Google Scholar]
  11. SIMPSON F. J., McCOY E. The amylases of five streptomycetes. Appl Microbiol. 1953 Sep;1(5):228–236. doi: 10.1128/am.1.5.228-236.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. STARK E., TETRAULT P. A. Isolation of bacterial, cell-free, starch saccharifying enzymes from the medium at 70 C. J Bacteriol. 1951 Aug;62(2):247–249. doi: 10.1128/jb.62.2.247-249.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wang H. L., Hesseltine C. W. Studies on the extracellular proteolytic enzymes of Rhizopus oligosporus. Can J Microbiol. 1965 Aug;11(4):727–732. doi: 10.1139/m65-096. [DOI] [PubMed] [Google Scholar]

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