Correlation between microbial protein thermostability and resistance to denaturation in aqueous-organic solvent 2-phase systems

RK Owusu, DA Cowan

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

The thermostability of cell-free protein extracts (from 10 microorganisms grown at 37–84°C) and of 10 fully or partially purified neutral proteases was determined in aqueous solution (Tris-HCl buffer 0.1 m. pH 7.0) using protein solubility and enzyme activity loss, respectively, as the criteria of denaturation. The stability of the protein extracts and purified proteins was also determined in an aqueous-water immiscible organic solvent two-phase system using the same criteria. There was a strong positive correlation between microorganism growth temperature and the thermostability of protein extracts in both aqueous and aqueous:organic two-phase systems. A correlation also existed between protease thermostability and the stability in the aqueous:organic two-phase system. Protein stability was higher in those aqueous:organic two-phase systems with a more hydrophobic organic phase. However, for pairs of organic solvents of similar hydrophobicity, the aqueous:organic two-phase system with the higher rated interfacial tension was more denaturing. Protein extract stability was sensitive to both aqueous:gas and aqueous:organic interface size depending on the nature of the organic phase.
LanguageEnglish
Pages568-574
JournalEnzyme and Microbial Technology
Volume11
Issue number9
DOIs
Publication statusPublished - 1 Sep 1989

Fingerprint

Denaturation
Organic solvents
Proteins
Microorganisms
Peptide Hydrolases
Enzyme activity
Growth temperature
Hydrophobicity
Surface tension
Buffers
Solubility
Gases
Water

Keywords

  • Thermophilic proteins
  • protein thermostability: protein stability
  • aqueous:organic solvent two-phase systems

Cite this

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title = "Correlation between microbial protein thermostability and resistance to denaturation in aqueous-organic solvent 2-phase systems",
abstract = "The thermostability of cell-free protein extracts (from 10 microorganisms grown at 37–84°C) and of 10 fully or partially purified neutral proteases was determined in aqueous solution (Tris-HCl buffer 0.1 m. pH 7.0) using protein solubility and enzyme activity loss, respectively, as the criteria of denaturation. The stability of the protein extracts and purified proteins was also determined in an aqueous-water immiscible organic solvent two-phase system using the same criteria. There was a strong positive correlation between microorganism growth temperature and the thermostability of protein extracts in both aqueous and aqueous:organic two-phase systems. A correlation also existed between protease thermostability and the stability in the aqueous:organic two-phase system. Protein stability was higher in those aqueous:organic two-phase systems with a more hydrophobic organic phase. However, for pairs of organic solvents of similar hydrophobicity, the aqueous:organic two-phase system with the higher rated interfacial tension was more denaturing. Protein extract stability was sensitive to both aqueous:gas and aqueous:organic interface size depending on the nature of the organic phase.",
keywords = "Thermophilic proteins, protein thermostability: protein stability, aqueous:organic solvent two-phase systems",
author = "RK Owusu and DA Cowan",
note = "Reference text: 1. AHERN TJ THE MECHANISM OF IRREVERSIBLE ENZYME INACTIVATION AT 100-DEGREES-C SCIENCE 228 : 1280 1985 2. AMELUNXEN RE MECHANISMS OF THERMOPHILY CRC CRITICAL REVIEWS IN MICROBIOLOGY 6 : 343 1978 3. ANTONINI E ENZYME CATALYZED-REACTIONS IN WATER-ORGANIC SOLVENT 2-PHASE SYSTEMS ENZYME AND MICROBIAL TECHNOLOGY 3 : 291 1981 4. BRINK LES OPTIMIZATION OF ORGANIC-SOLVENT IN MULTIPHASE BIOCATALYSIS BIOTECHNOLOGY AND BIOENGINEERING 27 : 1258 1985 5. BROCK TD LIFE AT HIGH-TEMPERATURES SCIENCE 230 : 132 1985 6. BUTLER LG ENZYMES IN NON-AQUEOUS SOLVENTS ENZYME AND MICROBIAL TECHNOLOGY 1 : 253 1979 7. CARREA G BIOCATALYSIS IN WATER-ORGANIC SOLVENT 2-PHASE SYSTEMS TRENDS IN BIOTECHNOLOGY 2 : 102 1984 8. CLAUS D DTSCH SAMMLUNG MIKRO : 1983 9. COWAN DA PURIFICATION AND SOME PROPERTIES OF AN EXTRACELLULAR PROTEASE (CALDOLYSIN) FROM AN EXTREME THERMOPHILE BIOCHIMICA ET BIOPHYSICA ACTA 705 : 293 1982 10. DANIEL RM A CORRELATION BETWEEN PROTEIN THERMOSTABILITY AND RESISTANCE TO PROTEOLYSIS BIOCHEMICAL JOURNAL 207 : 641 1982 11. DAVIES JT INTERFACIAL PHENOMEN : 1 1961 12. DONALDSON TL KINETICS OF PROTEIN DENATURATION AT GAS-LIQUID INTERFACES JOURNAL OF COLLOID AND INTERFACE SCIENCE 74 : 441 1980 13. FRANKS F CRC CRIT REV BIOCHEM 3 : 165 1975 14. FRENDSDORFF HK THE KINETICS OF PROTEIN DENATURATION .4. THE VISCOSITY AND GELATION OF UREA SOLUTIONS OF OVALBUMIN JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 75 : 5157 1953 15. FUKUSHIMA D DENATURATION OF SOYBEAN PROTEINS BY ORGANIC SOLVENTS CEREAL CHEMISTRY 46 : 156 1969 16. GRAHAM DE PROTEINS AT LIQUID INTERFACES .3. MOLECULAR-STRUCTURES OF ADSORBED FILMS JOURNAL OF COLLOID AND INTERFACE SCIENCE 70 : 427 1979 17. HICKEY CW ELECTRON-TRANSPORT SYSTEM OF AN EXTREMELY THERMOPHILIC BACTERIUM JOURNAL OF GENERAL MICROBIOLOGY 114 : 195 1979 18. HOLME J THERMAL DENATURATION AND AGGREGATION OF OVALBUMIN JOURNAL OF PHYSICAL CHEMISTRY 67 : 782 1963 19. HORBETT TA PROTEINS INTERFACES : 1 1987 20. HUDSON JA ISOLATION AND CHARACTERIZATION OF A NEW CALDOACTIVE FILAMENTOUS BACTERIUM FEMS MICROBIOLOGY LETTERS 22 : 149 1984 21. JAMES LK ADSORPTION OF ENZYMES AT INTERFACES - FILM FORMATION AND EFFECT ON ACTIVITY ADVANCES IN ENZYMOLOGY AND RELATED AREAS OF MOLECULAR BIOLOGY 28 : 1 1966 22. JOLY M PHYSICO CHEM APPROAC : 1965 23. KHMELNITSKY YL DETERGENTLESS MICROEMULSIONS - A NEW MICROHETEROGENEOUS MEDIUM FOR ENZYMATIC-REACTION ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 501 : 161 1987 24. KLIBANOV AM STABILIZATION OF ENZYMES AGAINST THERMAL INACTIVATION ADVANCES IN APPLIED MICROBIOLOGY 29 : 1 1983 25. KOFFLER H THE RELATIVE THERMOSTABILITY OF CYTOPLASMIC PROTEINS FROM THERMOPHILIC BACTERIA ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 67 : 249 1957 26. LAANE C BIOCATALYSIS ORGANIC : 65 1987 27. LILLY MD 2-LIQUID-PHASE BIOCATALYTIC REACTIONS JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY 32 : 162 1982 28. MOZHAEV VV STRUCTURE STABILITY RELATIONSHIPS IN PROTEINS - NEW APPROACHES TO STABILIZING ENZYMES ENZYME AND MICROBIAL TECHNOLOGY 6 : 50 1984 29. OSHIMA T ENZYME ENG 4 : 41 1978 30. PETERSON GL SIMPLIFICATION OF PROTEIN ASSAY METHOD OF LOWRY ET AL - WHICH IS MORE GENERALLY APPLICABLE ANALYTICAL BIOCHEMISTRY 83 : 346 1977 31. ROTHERN A ADV PROTEIN CHEM 3 : 123 1948 32. SCHELLMAN JA SOLVENT DENATURATION BIOPOLYMERS 17 : 1305 1978 33. SCHRIER EE EFFECT OF AQUEOUS ALCOHOL SOLUTIONS ON THERMAL TRANSITION OF RIBONUCLEASE JOURNAL OF PHYSICAL CHEMISTRY 69 : 298 1965 34. SINGER SJ THE PROPERTIES OF PROTEINS IN NONAQUEOUS SOLVENTS ADVANCES IN PROTEIN CHEMISTRY 17 : 1 1962 35. SODA K TRENDS BIOCHEM SCI 8 : 428 1987 36. SOKAL RR INTRO BIOSTATISTICS : 260 1973 37. SUZUKI C GELATION OF OVALBUMIN SOLUTIONS BY HIGH PRESSURES ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 102 : 367 1963 38. TANFORD C ADV PROTEIN CHEM 23 : 121 1968 39. ZAKS A J BIOL CHEM 236 : 1988 40. ZALE SE WHY DOES RIBONUCLEASE IRREVERSIBLY INACTIVATE AT HIGH-TEMPERATURES BIOCHEMISTRY 25 : 5432 1986 41. ZALE SE ON THE ROLE OF REVERSIBLE DENATURATION (UNFOLDING) IN THE IRREVERSIBLE THERMAL INACTIVATION OF ENZYMES BIOTECHNOLOGY AND BIOENGINEERING 25 : 2221 1983",
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}

Correlation between microbial protein thermostability and resistance to denaturation in aqueous-organic solvent 2-phase systems. / Owusu, RK; Cowan, DA.

In: Enzyme and Microbial Technology, Vol. 11, No. 9, 01.09.1989, p. 568-574.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Correlation between microbial protein thermostability and resistance to denaturation in aqueous-organic solvent 2-phase systems

AU - Owusu, RK

AU - Cowan, DA

N1 - Reference text: 1. AHERN TJ THE MECHANISM OF IRREVERSIBLE ENZYME INACTIVATION AT 100-DEGREES-C SCIENCE 228 : 1280 1985 2. AMELUNXEN RE MECHANISMS OF THERMOPHILY CRC CRITICAL REVIEWS IN MICROBIOLOGY 6 : 343 1978 3. ANTONINI E ENZYME CATALYZED-REACTIONS IN WATER-ORGANIC SOLVENT 2-PHASE SYSTEMS ENZYME AND MICROBIAL TECHNOLOGY 3 : 291 1981 4. BRINK LES OPTIMIZATION OF ORGANIC-SOLVENT IN MULTIPHASE BIOCATALYSIS BIOTECHNOLOGY AND BIOENGINEERING 27 : 1258 1985 5. BROCK TD LIFE AT HIGH-TEMPERATURES SCIENCE 230 : 132 1985 6. BUTLER LG ENZYMES IN NON-AQUEOUS SOLVENTS ENZYME AND MICROBIAL TECHNOLOGY 1 : 253 1979 7. CARREA G BIOCATALYSIS IN WATER-ORGANIC SOLVENT 2-PHASE SYSTEMS TRENDS IN BIOTECHNOLOGY 2 : 102 1984 8. CLAUS D DTSCH SAMMLUNG MIKRO : 1983 9. COWAN DA PURIFICATION AND SOME PROPERTIES OF AN EXTRACELLULAR PROTEASE (CALDOLYSIN) FROM AN EXTREME THERMOPHILE BIOCHIMICA ET BIOPHYSICA ACTA 705 : 293 1982 10. DANIEL RM A CORRELATION BETWEEN PROTEIN THERMOSTABILITY AND RESISTANCE TO PROTEOLYSIS BIOCHEMICAL JOURNAL 207 : 641 1982 11. DAVIES JT INTERFACIAL PHENOMEN : 1 1961 12. DONALDSON TL KINETICS OF PROTEIN DENATURATION AT GAS-LIQUID INTERFACES JOURNAL OF COLLOID AND INTERFACE SCIENCE 74 : 441 1980 13. FRANKS F CRC CRIT REV BIOCHEM 3 : 165 1975 14. FRENDSDORFF HK THE KINETICS OF PROTEIN DENATURATION .4. THE VISCOSITY AND GELATION OF UREA SOLUTIONS OF OVALBUMIN JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 75 : 5157 1953 15. FUKUSHIMA D DENATURATION OF SOYBEAN PROTEINS BY ORGANIC SOLVENTS CEREAL CHEMISTRY 46 : 156 1969 16. GRAHAM DE PROTEINS AT LIQUID INTERFACES .3. MOLECULAR-STRUCTURES OF ADSORBED FILMS JOURNAL OF COLLOID AND INTERFACE SCIENCE 70 : 427 1979 17. HICKEY CW ELECTRON-TRANSPORT SYSTEM OF AN EXTREMELY THERMOPHILIC BACTERIUM JOURNAL OF GENERAL MICROBIOLOGY 114 : 195 1979 18. HOLME J THERMAL DENATURATION AND AGGREGATION OF OVALBUMIN JOURNAL OF PHYSICAL CHEMISTRY 67 : 782 1963 19. HORBETT TA PROTEINS INTERFACES : 1 1987 20. HUDSON JA ISOLATION AND CHARACTERIZATION OF A NEW CALDOACTIVE FILAMENTOUS BACTERIUM FEMS MICROBIOLOGY LETTERS 22 : 149 1984 21. JAMES LK ADSORPTION OF ENZYMES AT INTERFACES - FILM FORMATION AND EFFECT ON ACTIVITY ADVANCES IN ENZYMOLOGY AND RELATED AREAS OF MOLECULAR BIOLOGY 28 : 1 1966 22. JOLY M PHYSICO CHEM APPROAC : 1965 23. KHMELNITSKY YL DETERGENTLESS MICROEMULSIONS - A NEW MICROHETEROGENEOUS MEDIUM FOR ENZYMATIC-REACTION ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 501 : 161 1987 24. KLIBANOV AM STABILIZATION OF ENZYMES AGAINST THERMAL INACTIVATION ADVANCES IN APPLIED MICROBIOLOGY 29 : 1 1983 25. KOFFLER H THE RELATIVE THERMOSTABILITY OF CYTOPLASMIC PROTEINS FROM THERMOPHILIC BACTERIA ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 67 : 249 1957 26. LAANE C BIOCATALYSIS ORGANIC : 65 1987 27. LILLY MD 2-LIQUID-PHASE BIOCATALYTIC REACTIONS JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY 32 : 162 1982 28. MOZHAEV VV STRUCTURE STABILITY RELATIONSHIPS IN PROTEINS - NEW APPROACHES TO STABILIZING ENZYMES ENZYME AND MICROBIAL TECHNOLOGY 6 : 50 1984 29. OSHIMA T ENZYME ENG 4 : 41 1978 30. PETERSON GL SIMPLIFICATION OF PROTEIN ASSAY METHOD OF LOWRY ET AL - WHICH IS MORE GENERALLY APPLICABLE ANALYTICAL BIOCHEMISTRY 83 : 346 1977 31. ROTHERN A ADV PROTEIN CHEM 3 : 123 1948 32. SCHELLMAN JA SOLVENT DENATURATION BIOPOLYMERS 17 : 1305 1978 33. SCHRIER EE EFFECT OF AQUEOUS ALCOHOL SOLUTIONS ON THERMAL TRANSITION OF RIBONUCLEASE JOURNAL OF PHYSICAL CHEMISTRY 69 : 298 1965 34. SINGER SJ THE PROPERTIES OF PROTEINS IN NONAQUEOUS SOLVENTS ADVANCES IN PROTEIN CHEMISTRY 17 : 1 1962 35. SODA K TRENDS BIOCHEM SCI 8 : 428 1987 36. SOKAL RR INTRO BIOSTATISTICS : 260 1973 37. SUZUKI C GELATION OF OVALBUMIN SOLUTIONS BY HIGH PRESSURES ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 102 : 367 1963 38. TANFORD C ADV PROTEIN CHEM 23 : 121 1968 39. ZAKS A J BIOL CHEM 236 : 1988 40. ZALE SE WHY DOES RIBONUCLEASE IRREVERSIBLY INACTIVATE AT HIGH-TEMPERATURES BIOCHEMISTRY 25 : 5432 1986 41. ZALE SE ON THE ROLE OF REVERSIBLE DENATURATION (UNFOLDING) IN THE IRREVERSIBLE THERMAL INACTIVATION OF ENZYMES BIOTECHNOLOGY AND BIOENGINEERING 25 : 2221 1983

PY - 1989/9/1

Y1 - 1989/9/1

N2 - The thermostability of cell-free protein extracts (from 10 microorganisms grown at 37–84°C) and of 10 fully or partially purified neutral proteases was determined in aqueous solution (Tris-HCl buffer 0.1 m. pH 7.0) using protein solubility and enzyme activity loss, respectively, as the criteria of denaturation. The stability of the protein extracts and purified proteins was also determined in an aqueous-water immiscible organic solvent two-phase system using the same criteria. There was a strong positive correlation between microorganism growth temperature and the thermostability of protein extracts in both aqueous and aqueous:organic two-phase systems. A correlation also existed between protease thermostability and the stability in the aqueous:organic two-phase system. Protein stability was higher in those aqueous:organic two-phase systems with a more hydrophobic organic phase. However, for pairs of organic solvents of similar hydrophobicity, the aqueous:organic two-phase system with the higher rated interfacial tension was more denaturing. Protein extract stability was sensitive to both aqueous:gas and aqueous:organic interface size depending on the nature of the organic phase.

AB - The thermostability of cell-free protein extracts (from 10 microorganisms grown at 37–84°C) and of 10 fully or partially purified neutral proteases was determined in aqueous solution (Tris-HCl buffer 0.1 m. pH 7.0) using protein solubility and enzyme activity loss, respectively, as the criteria of denaturation. The stability of the protein extracts and purified proteins was also determined in an aqueous-water immiscible organic solvent two-phase system using the same criteria. There was a strong positive correlation between microorganism growth temperature and the thermostability of protein extracts in both aqueous and aqueous:organic two-phase systems. A correlation also existed between protease thermostability and the stability in the aqueous:organic two-phase system. Protein stability was higher in those aqueous:organic two-phase systems with a more hydrophobic organic phase. However, for pairs of organic solvents of similar hydrophobicity, the aqueous:organic two-phase system with the higher rated interfacial tension was more denaturing. Protein extract stability was sensitive to both aqueous:gas and aqueous:organic interface size depending on the nature of the organic phase.

KW - Thermophilic proteins

KW - protein thermostability: protein stability

KW - aqueous:organic solvent two-phase systems

U2 - 10.1016/0141-0229(89)90084-7

DO - 10.1016/0141-0229(89)90084-7

M3 - Article

VL - 11

SP - 568

EP - 574

JO - Enzyme and Microbial Technology

T2 - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

IS - 9

ER -