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Effect of detergents on enzymic metabolism in microorganisms. A review
Zbl. Mikrobiol. 137 (1982), 42-45 Chemical Laboratories, Allahabad University, Allahabad, India]
Effect of Detergents on Enzymic Metabolism in Microorganisms. A Review VIJAY SHANKAR TRIPATHI, DINSHAW SORABJI (late), and PARVEEN TRIPATHI Detergents are fundamentally linear molecules, one end of which has a large non-polar hydrocarbon chain which is oil-soluble, while the other end is polar and water-soluble. These complex molecules exert far-reaching effects on living systems - the most pr imitive form being microorganisms. SIM and SIM (1979) found that intrinsic membrane protein hydrogenase of Paracoccus denitrijicans was solubilized on addition of Triton X-100. The partial specific volume of solubilized hydrogenase in the presence and absence of Triton X-100 was found to be 0.73 and 0.74 mlfg, respectively, indicating that the hydrogenase was much less bound than a miscelle of Triton X-100. Sedimentation coefficient increased from 10.4 S to 15.9 S when the detergent was removed, while Stokes radius of the enzyme in presence of Triton X-lOO, using gel filtration on Sepharose 6 B, was found to be 5.5 nm, but in absence of detergent the radius was 6.5 nm. Apparent molecular weight, on removing the detergent, increased from 242,500 to 466,000. Thus, hydrogenase behaved as a monocovalently linked tetramer in the presence of Triton X-100, while in its absence associated to form octamer. KARIBIAN (1973) observed a loss of oxidase activity of dihydrorotate dehydrogenase with the electron acceptor dichloroindophenol when nonionic detergents were treated with enzyme, obtained from Escherichia coli K 12 . The Triton X-lOO did not modify the K m app. of the enzyme, but orotic acid, the inhibition by-product, changed from a mixed to a competitive type. The enzyme solubilized, simultaneously the particle size decreased and the density was lowered, while the enzyme was stimulated by Triton X-lOO. Treatment of phospholipase A2 gave loss of activity, probably due to inhibition by the fatty acids and partly to the loss of the phospholipids. This activity was restored when Triton X-100 was inducted in the reaction mixture. YUAN-CHI Su and YUAN-HwANG SHU (1973) found that the enzymic activity of alkaline phosphatase, isolated from Bacillus species,was not affected in the presence of 1.5 % solution of Triton X-lOO. MATHUR and KEENAN (1974) isolated alkaline phosphatase from milk fat globule membrane and Escherichia COll: and observed unusual stability in presence of sodium dodecylsulphate and good retention of activity, even in a wide range of the detergent concentrations. NYBEBG (1979) studied the alkaline and acid phosphatase activities and phosphate accumulation in Nitzschia actinastroides in presence of sodium dodecyl sulphate, Triton X-100, sodium deoxycholate, and acetyltrimethyl ammonium bromide. The effect of phosphate starvation on the phosphatase activity was assessed, and it was found that detergents enhanced the phosphatase activities, while at low concentration of detergent the activity was markedly increased, but at higher concentrations the activities were found to greatly decrease. Presence of detergents lowered the amount of extractable phosphorus in the cells. J OLOHINE and REISS-HusSON (1974) prepared reaction centres and incubated round a wild type of Rhodopseudomonae spheroides Y with acetyltrimethyl ammonium bromide and found that the reaction centres contained lipids with only halfthe amount of phospholipid, since the rest was replaced by detergent. They got a phospholipid
Effect of Detergents on Enzymic Metabolism
43
with Rr, similar to diphosphatidyl glycerol, along with phosphatidyl ethanol amine, phosphatidyl glycerol, and phosphatidyl choline. PALAMARCZYK et al. (1979) found that presence of polyprenyl phosphate protected the membrane fractions, containing glycotransferase of Saccharomyces cerevisiae. When yeast membrane was incubated with Triton X-IOO or Nonidet P-40, it was found to yield N-acetyl glucosamine transferase which inhibited transfer of N-acetyl glucosamine-Pf) from UDP-Naoetylglucosamine-PC to the endogenous membrane lipids. The presence of dolichylphosphates in the incubating medium maintained the activity by acting as an acceptor for N-acetyl glucosamine. This protective effect was markedly absent when polyprenyl phosphates were added after pre-incubation of the detergent in the reaction mixture. FORSTEN and KARLJALAINEN (1977) observed complete inhibition with Oa(OH)2 solution, while there was no effect on the activity of arylamino peptidase of carious dentin by detergent. VAITKEVICIUS (1978) decomposed Bacillus subtilis cells, centrifuged, and obtained aminopeptidase from the soluble fraction at a pH range from 5.8-6.5 in presence of trimethyl ammonium salts; the yield was found to be greatly enhanced. SUSMAN and HAYS (1977) observed that detergents irreversibly inactivated membrane-bound enzyme IlIac, which was the membrane-bound enzyme of lactose transferase of Staphylococcus aureus, Triton X-IOO of 1.0 % concentration and at 25 °0 converted the enzyme into particulate and active form. Detergent concentration of 0.5 % solubilized the enzyme with high retention activity, while when the temperature of the system was varied to 37 °0 it resulted in irreversible and rapid inactivation. BOYER (1973) prepared biosynthetic systems, containing feedstock materials and their specifically active microorganisms, selected for their ability to produce a specific enzyme. The maximum activity of the enzymes was obtained by fermenting the microorganisms in a common nutrient solution, containing nitrogen compounds and minerals. LANYI (1973) studied the electron transport on interaction between the membrane lipids and Triton X-IOOin Halobacterium cutirubrum and observed no resistance when the respiration-inhibited cells were lysed. The absorption spectra of Bacterio ruberin showed possible association of this pigment with Triton X-IOO in presence of KNOO-inhibited cells. The integrity of cell-wall envelope was lost, along with resistance to penetration of the membrane cell envelopes by Triton X-lOO. This was apparently due to resistance to perturbation. The release of various intracellular components occurred along with menodione reductase activity. NAKAMURA et al. (1973) studied the lysis of Micrococcus radiodurans by cooperative action of nonionic detergents with the bacteriolytic enzyme of Achromobacter lyticus and found Triton X-IOO to be most effective in initiation of immediate lysis of the cells which, on washing with buffer and centrifuging, was easily used with the enzyme alone and with lysozyme. TRIPATHI and TRIPATHI (1980) reviewed detergent effect on metabolic changes in microorganisms. ONIYANAGI et al. (1976) prepared a mold bran of Aspergillus niger, IFO 4414, extracted with water. The enzyme solution, containing I 8130 and II 5640 units, was mixed with 0.1 % of pH 4.8 dodecyl sulphate at 70 °0 for 15 min and the residual activities were found to be 88.55 % and 2.61 %, respectively. SUGIRA (1977) obtained 1,500 ml solution of crude lipase from Ohromobacterium vicosum of sp. activity 52, yielding pure lipase of sp. activity 4,300. STARKOVA and STARK~ (1978) found that morphological mon mutants and a chain-forming env 0 mutant of Escherichia coli K 12 were hypersensitive to sodium dodecylsulphate and sodium deoxycholate. Employment of electron microscopy revealed that the detergents dissolved the cytoplasmic membrane of growing mon and env 0 cells, resulting
44
V. S.
TRIPATHI, D. SORABJI (late), and P. TRIPATHI
in dissociation of cytoplasm into two parts of unequal d. The mutant env C was lysed by lysozyme in absence of EDTA, and absence of osmotic shock was found to release alkaline phosphatase, and was sensitive to actinomycin D and riframpicin, revealing a gross perturbation of the outer membrane. LAHAU et al. (1979) investigated the bactericidal and bacteriolytic effects of lysolecithin and egg white lysozyme on Staphylococcus aureus and group A streptococci, along with solubilization of phospholipids from the bacterial membrane. They found that low concentrations of lysolecithin, i.e. at 1-10,ug/h., were highly bactericidal for Staphylococcus aureus and group A streptococci, but did not induce bacteriolysis or solubilization of a substantial amount of phospholipids. Lysolecithin at 50,ug/ml caused substantial lipid release from staphylococci, while lysozyme + lysolecithin were essential for solubilizing lipids from streptococci. They found that the release of lipids by lysozyme from group A streptococci was related to its enzymic activity on an unknown substrate, and not its cationic nature, as the muramidase could not be replaced by cationic substances. Polyelectrolytes did not inhibit the release of lipids from staphylococci, while they induced lipid release from group A streptococci. JENSEN (1973) tested the effectiveness of various detergents on the extra-cellular proteases, obtained from Bacillus strains. YOBE et al. (1973) cultured Bacillus species FERH-P-1396 in a nutrient medium at pH 8 and 34°C, and an alkaline proteinase of mol. wt. 15,000-20,000, stable at pH 5-11, was obtained. WILLETS and CAIN (1972) studied the metabolism of A. bacillus with alkylbenzene sulphonates and found initial oxidation of the terminal methyl group and release of desulphonating enzyme-cytochrome reductase (E.C. 1.1.3.1), detectable in cell-free extracts. The alkyl side chain, after oxidation of the terminal methyl group, underwent §-oxidation, and acylcoenzyme synthetase (E.C. 6.2.1.3) was identified in cellfree extracts. RUSSEL (1979) used Triton X-I00 to overcome the inhibition by sodium dodecyl sulphate of fructosyl transferase from Streptococcus mutans. MARKHAM and JANES (1978) investigated immunological and pathological response of aerosolized bacterial protease and detergent. TAMORI et al. (1979) prepared crude membranebound phospholipase which was detergent-resistant from E. coli K-12. The phospholipase W80S stable and active even on incubation at 100°C, but, when highly purified with removal of Triton X-lOO, was found to be strikingly thermolabile. The inactivation by Triton X-I00 was due to a biphasic temperature dependence, i.e. at 37°C and above 70 °C, at the latter temperature the electrophoretic mobility was found to change, while at 37°C Triton X-lOO effectively protected the enzyme. CHRISTIANSEN and KILLIAN (1977) studied the effect of chlorhexidine glueonate and some detergents on activity of dextransucrase of Streptoocccus mutans and found it to be inhibitory.
References BOYER, L. D.: Concurrent production of a plurality of enzymes. U.S. 3, 769, 167/c1. 195/66R (CO 7 g) 30 Oct. 1973, Appl, 148, 256, 24/5/1971, 8 pp. CHRISTIANSEN, F., and KILLIAN, M.: The effect of chlorhexidine and some other detergents on the activity of dextransucrase from Streptococcus muiane. Acta Odontal. Scand. 35 (1977) (3), 119-123. FORSTEN, L., and KARI,JAIAINEN, S.: Effect of Ca(OHh solution and a chlorohexidir::e-basec1 detergent on the microbial activity. Acta Odontal, Scand. 35 (1977) (6), 275-280. JENSEN, G.: Development of new improved enzymes for application in washing agents. Fette, Seifen, Anstrichmitte175 (1973) (I), 48-55. JOLCHINE, G., and REISS-HusSON, F.: Comparative studies on two reaction centre preparations from Iihodopseudomonas spheroides, Fed. Eur. Biochem. Soc. Lett. 40 (1974) (1), 5-8. KARIBIAN, D.: Dihydrorotate dehydrogenase of E. coli K 2 effects of Triton X-I00 and phospholipids. Biochim. Biophys. Acta 302 (1973) (2), 205 - 215.
Effect of Detergents on Enzymic Metabolism
45
LANYI, J. K.: Influence of electron transport on the interaction between membrane lipids and Triton X-I00 in Halobacterium cutir'ubrum. Biochemistry 12 (1973) (2), 1433-1438. LAHAU,M., NEELMAN, N. SELA,M. N., and GINSBERG,!.: Effect ofleukocyte hydrolysis on bacteria. XIII. Role played by leukocyte extracts, lecithin, phospholipase A 2, lysozyme cationic proteins, and detergents in the solubilization of lipids from Staphylococcu8 aureus and group A streptococci. Inflammation (N.Y.) a (1979) (4), 365-377. MARKHAM, J., and JAMES, F.: Immunologic and pathologic response to aerosolized bacterial protease and detergent. Natl. Libr. Canada, Ottawa, Onto Diss. Abstr, Int. B 38 (1978) (10), 4676. MATHUR,1. H., and KEENAN, T. W.: Stability of alkaline phosphatase in sodium dodecylsulphate. FEES Let. 44 (1974) (1), 79-82. NAKAMURA, K., OKAZAWA, Y., SOEJIMA, M., and MASAKI, T.: Lysis of Micrococcus radiodurans. Agr. BioI. Chem. 3 (1973). NYBERG, H.: The effects of some detergents on the phosphatases and phosphate accumulation of Nitzschia actiruistroides, Ann. Bot. Fenn. 16 (1979) (1), 28-34. ONIYANAGI, T" FUG]SJiIMA, T., CCHIDA, K., and YOSHINO, H.: 5' Phosphodiesterase. Japan Kokai 77, 136, 989 (Cl. Cl. 2 D 13/10) 16.11. 1977, Appl. 76/52, 228, 10.5. 1976. 6 pp. PALAMARCZYK, G., LEHLE, L., and TANNER, W.: Polyprenylphosphate prevents inactivation of yeast glycotransferase by detergents. FEBS. Lett. 108 (1979) (1), 111-115. RUSSEL, R. R. B.: Use of Triton X-I00 to overcome the inhibition of fructosyltransferase by S.D.S. Anal. Biochem. 97 (1979) (I), 173-175. SHiI, E., and SIM, R. B.: Hydrodynamic parameters of the detergent solubilized hydrogenase from Pnracoccue denitr(ficans. Eur. J. Biochem. 97 (1979) (1), 119 -126. STARKOVA, Z., and STARKA, J.: Morphological mutants of E8cherichia coli: Nature of the permeability barrier in mon and env C cells. Ann. Microbiol. (Paris) 129 A (1978) (3), 265-284. SUGIRA, M., IsOBE, M.: Japan Kokai 7796, 790 (Cl. C 07 G 7)/02) 13th Aug. 1977. Appl. 76/11, 846, 07 Feb. 1976 4 pp. SUSMAN, M. L., and HAYS, J. B.: Irreversible inactivation of membrane-bound enzyme in II lac of the lactose phosphotransferase system of Staphylococcus aureus by Triton X-I00 and protection by substrates. Biochim. Biophys. Acta 465 (1977) (3), 559-570. TAMORI, Y., NISHIJIMO, M., and NOJIMA, S.: Properties of purified detergent-resistant phospholipase A of Escherichia coli K-I2. Inactivation and protection with detergents and phospholipids. TRIPATHI, V. S., and TRIPATHI, P.: Detergent effect on metabolic changes in microorganisms. A Review. Zbl. Bakt. II 135 (1980), 510-514. VAITKIRCIUS, R., CIPENS, L., MASIULYTE, G., and ANTONEKOVA, L. lVI.: Separation of intracellular amino peptidase from Bacillus eubtilie, Ortkrytiya, Izobret., Prom. Obraztsy, Tovarnye Znaki 55 (1978) (125), 102. VVILLETS, A. J.: Microbial metabolism of alkyl-benzene sulphonates. Fungal metabolism of 1phenylundecane-p-sulphonate and I-phenyldodecane-p-sulphonate. Antonie von Leeuwenhoek 39 (1976) (4), 567 -585. YOBE, S., MIROSE, Y., NAKAMURA, Y., and MITSUJI, K.: Alkaline proteinase production by bacterial fermentation. Ger. Offen-2, 313, 546 (C!. C. 12. d - CO 7 g), 27 Sept. 1973. Japan Appl. 72/27 G 73, March 18, 1972 , 15 pp. YUAN-CHI Su and SHu-YANU HWANG: Alkaline protease produced by Bacillus species. J. chin. Biochem. Soc. 2 (1973) (1), 1-10. Author's address: VIJAY SHANKAR TRIPATHI, Chemical Laboratories, Allahabad University, Allahabad, India.
Effect of Detergents on Enzymic Metabolism in Microorganisms. A Review VIJAY SHANKAR TRIPATHI, DINSHAW SORABJI (late), and PARVEEN TRIPATHI Detergents are fundamentally linear molecules, one end of which has a large non-polar hydrocarbon chain which is oil-soluble, while the other end is polar and water-soluble. These complex molecules exert far-reaching effects on living systems - the most pr imitive form being microorganisms. SIM and SIM (1979) found that intrinsic membrane protein hydrogenase of Paracoccus denitrijicans was solubilized on addition of Triton X-100. The partial specific volume of solubilized hydrogenase in the presence and absence of Triton X-100 was found to be 0.73 and 0.74 mlfg, respectively, indicating that the hydrogenase was much less bound than a miscelle of Triton X-100. Sedimentation coefficient increased from 10.4 S to 15.9 S when the detergent was removed, while Stokes radius of the enzyme in presence of Triton X-lOO, using gel filtration on Sepharose 6 B, was found to be 5.5 nm, but in absence of detergent the radius was 6.5 nm. Apparent molecular weight, on removing the detergent, increased from 242,500 to 466,000. Thus, hydrogenase behaved as a monocovalently linked tetramer in the presence of Triton X-100, while in its absence associated to form octamer. KARIBIAN (1973) observed a loss of oxidase activity of dihydrorotate dehydrogenase with the electron acceptor dichloroindophenol when nonionic detergents were treated with enzyme, obtained from Escherichia coli K 12 . The Triton X-lOO did not modify the K m app. of the enzyme, but orotic acid, the inhibition by-product, changed from a mixed to a competitive type. The enzyme solubilized, simultaneously the particle size decreased and the density was lowered, while the enzyme was stimulated by Triton X-lOO. Treatment of phospholipase A2 gave loss of activity, probably due to inhibition by the fatty acids and partly to the loss of the phospholipids. This activity was restored when Triton X-100 was inducted in the reaction mixture. YUAN-CHI Su and YUAN-HwANG SHU (1973) found that the enzymic activity of alkaline phosphatase, isolated from Bacillus species,was not affected in the presence of 1.5 % solution of Triton X-lOO. MATHUR and KEENAN (1974) isolated alkaline phosphatase from milk fat globule membrane and Escherichia COll: and observed unusual stability in presence of sodium dodecylsulphate and good retention of activity, even in a wide range of the detergent concentrations. NYBEBG (1979) studied the alkaline and acid phosphatase activities and phosphate accumulation in Nitzschia actinastroides in presence of sodium dodecyl sulphate, Triton X-100, sodium deoxycholate, and acetyltrimethyl ammonium bromide. The effect of phosphate starvation on the phosphatase activity was assessed, and it was found that detergents enhanced the phosphatase activities, while at low concentration of detergent the activity was markedly increased, but at higher concentrations the activities were found to greatly decrease. Presence of detergents lowered the amount of extractable phosphorus in the cells. J OLOHINE and REISS-HusSON (1974) prepared reaction centres and incubated round a wild type of Rhodopseudomonae spheroides Y with acetyltrimethyl ammonium bromide and found that the reaction centres contained lipids with only halfthe amount of phospholipid, since the rest was replaced by detergent. They got a phospholipid
Effect of Detergents on Enzymic Metabolism
43
with Rr, similar to diphosphatidyl glycerol, along with phosphatidyl ethanol amine, phosphatidyl glycerol, and phosphatidyl choline. PALAMARCZYK et al. (1979) found that presence of polyprenyl phosphate protected the membrane fractions, containing glycotransferase of Saccharomyces cerevisiae. When yeast membrane was incubated with Triton X-IOO or Nonidet P-40, it was found to yield N-acetyl glucosamine transferase which inhibited transfer of N-acetyl glucosamine-Pf) from UDP-Naoetylglucosamine-PC to the endogenous membrane lipids. The presence of dolichylphosphates in the incubating medium maintained the activity by acting as an acceptor for N-acetyl glucosamine. This protective effect was markedly absent when polyprenyl phosphates were added after pre-incubation of the detergent in the reaction mixture. FORSTEN and KARLJALAINEN (1977) observed complete inhibition with Oa(OH)2 solution, while there was no effect on the activity of arylamino peptidase of carious dentin by detergent. VAITKEVICIUS (1978) decomposed Bacillus subtilis cells, centrifuged, and obtained aminopeptidase from the soluble fraction at a pH range from 5.8-6.5 in presence of trimethyl ammonium salts; the yield was found to be greatly enhanced. SUSMAN and HAYS (1977) observed that detergents irreversibly inactivated membrane-bound enzyme IlIac, which was the membrane-bound enzyme of lactose transferase of Staphylococcus aureus, Triton X-IOO of 1.0 % concentration and at 25 °0 converted the enzyme into particulate and active form. Detergent concentration of 0.5 % solubilized the enzyme with high retention activity, while when the temperature of the system was varied to 37 °0 it resulted in irreversible and rapid inactivation. BOYER (1973) prepared biosynthetic systems, containing feedstock materials and their specifically active microorganisms, selected for their ability to produce a specific enzyme. The maximum activity of the enzymes was obtained by fermenting the microorganisms in a common nutrient solution, containing nitrogen compounds and minerals. LANYI (1973) studied the electron transport on interaction between the membrane lipids and Triton X-IOOin Halobacterium cutirubrum and observed no resistance when the respiration-inhibited cells were lysed. The absorption spectra of Bacterio ruberin showed possible association of this pigment with Triton X-IOO in presence of KNOO-inhibited cells. The integrity of cell-wall envelope was lost, along with resistance to penetration of the membrane cell envelopes by Triton X-lOO. This was apparently due to resistance to perturbation. The release of various intracellular components occurred along with menodione reductase activity. NAKAMURA et al. (1973) studied the lysis of Micrococcus radiodurans by cooperative action of nonionic detergents with the bacteriolytic enzyme of Achromobacter lyticus and found Triton X-IOO to be most effective in initiation of immediate lysis of the cells which, on washing with buffer and centrifuging, was easily used with the enzyme alone and with lysozyme. TRIPATHI and TRIPATHI (1980) reviewed detergent effect on metabolic changes in microorganisms. ONIYANAGI et al. (1976) prepared a mold bran of Aspergillus niger, IFO 4414, extracted with water. The enzyme solution, containing I 8130 and II 5640 units, was mixed with 0.1 % of pH 4.8 dodecyl sulphate at 70 °0 for 15 min and the residual activities were found to be 88.55 % and 2.61 %, respectively. SUGIRA (1977) obtained 1,500 ml solution of crude lipase from Ohromobacterium vicosum of sp. activity 52, yielding pure lipase of sp. activity 4,300. STARKOVA and STARK~ (1978) found that morphological mon mutants and a chain-forming env 0 mutant of Escherichia coli K 12 were hypersensitive to sodium dodecylsulphate and sodium deoxycholate. Employment of electron microscopy revealed that the detergents dissolved the cytoplasmic membrane of growing mon and env 0 cells, resulting
44
V. S.
TRIPATHI, D. SORABJI (late), and P. TRIPATHI
in dissociation of cytoplasm into two parts of unequal d. The mutant env C was lysed by lysozyme in absence of EDTA, and absence of osmotic shock was found to release alkaline phosphatase, and was sensitive to actinomycin D and riframpicin, revealing a gross perturbation of the outer membrane. LAHAU et al. (1979) investigated the bactericidal and bacteriolytic effects of lysolecithin and egg white lysozyme on Staphylococcus aureus and group A streptococci, along with solubilization of phospholipids from the bacterial membrane. They found that low concentrations of lysolecithin, i.e. at 1-10,ug/h., were highly bactericidal for Staphylococcus aureus and group A streptococci, but did not induce bacteriolysis or solubilization of a substantial amount of phospholipids. Lysolecithin at 50,ug/ml caused substantial lipid release from staphylococci, while lysozyme + lysolecithin were essential for solubilizing lipids from streptococci. They found that the release of lipids by lysozyme from group A streptococci was related to its enzymic activity on an unknown substrate, and not its cationic nature, as the muramidase could not be replaced by cationic substances. Polyelectrolytes did not inhibit the release of lipids from staphylococci, while they induced lipid release from group A streptococci. JENSEN (1973) tested the effectiveness of various detergents on the extra-cellular proteases, obtained from Bacillus strains. YOBE et al. (1973) cultured Bacillus species FERH-P-1396 in a nutrient medium at pH 8 and 34°C, and an alkaline proteinase of mol. wt. 15,000-20,000, stable at pH 5-11, was obtained. WILLETS and CAIN (1972) studied the metabolism of A. bacillus with alkylbenzene sulphonates and found initial oxidation of the terminal methyl group and release of desulphonating enzyme-cytochrome reductase (E.C. 1.1.3.1), detectable in cell-free extracts. The alkyl side chain, after oxidation of the terminal methyl group, underwent §-oxidation, and acylcoenzyme synthetase (E.C. 6.2.1.3) was identified in cellfree extracts. RUSSEL (1979) used Triton X-I00 to overcome the inhibition by sodium dodecyl sulphate of fructosyl transferase from Streptococcus mutans. MARKHAM and JANES (1978) investigated immunological and pathological response of aerosolized bacterial protease and detergent. TAMORI et al. (1979) prepared crude membranebound phospholipase which was detergent-resistant from E. coli K-12. The phospholipase W80S stable and active even on incubation at 100°C, but, when highly purified with removal of Triton X-lOO, was found to be strikingly thermolabile. The inactivation by Triton X-I00 was due to a biphasic temperature dependence, i.e. at 37°C and above 70 °C, at the latter temperature the electrophoretic mobility was found to change, while at 37°C Triton X-lOO effectively protected the enzyme. CHRISTIANSEN and KILLIAN (1977) studied the effect of chlorhexidine glueonate and some detergents on activity of dextransucrase of Streptoocccus mutans and found it to be inhibitory.
References BOYER, L. D.: Concurrent production of a plurality of enzymes. U.S. 3, 769, 167/c1. 195/66R (CO 7 g) 30 Oct. 1973, Appl, 148, 256, 24/5/1971, 8 pp. CHRISTIANSEN, F., and KILLIAN, M.: The effect of chlorhexidine and some other detergents on the activity of dextransucrase from Streptococcus muiane. Acta Odontal. Scand. 35 (1977) (3), 119-123. FORSTEN, L., and KARI,JAIAINEN, S.: Effect of Ca(OHh solution and a chlorohexidir::e-basec1 detergent on the microbial activity. Acta Odontal, Scand. 35 (1977) (6), 275-280. JENSEN, G.: Development of new improved enzymes for application in washing agents. Fette, Seifen, Anstrichmitte175 (1973) (I), 48-55. JOLCHINE, G., and REISS-HusSON, F.: Comparative studies on two reaction centre preparations from Iihodopseudomonas spheroides, Fed. Eur. Biochem. Soc. Lett. 40 (1974) (1), 5-8. KARIBIAN, D.: Dihydrorotate dehydrogenase of E. coli K 2 effects of Triton X-I00 and phospholipids. Biochim. Biophys. Acta 302 (1973) (2), 205 - 215.
Effect of Detergents on Enzymic Metabolism
45
LANYI, J. K.: Influence of electron transport on the interaction between membrane lipids and Triton X-I00 in Halobacterium cutir'ubrum. Biochemistry 12 (1973) (2), 1433-1438. LAHAU,M., NEELMAN, N. SELA,M. N., and GINSBERG,!.: Effect ofleukocyte hydrolysis on bacteria. XIII. Role played by leukocyte extracts, lecithin, phospholipase A 2, lysozyme cationic proteins, and detergents in the solubilization of lipids from Staphylococcu8 aureus and group A streptococci. Inflammation (N.Y.) a (1979) (4), 365-377. MARKHAM, J., and JAMES, F.: Immunologic and pathologic response to aerosolized bacterial protease and detergent. Natl. Libr. Canada, Ottawa, Onto Diss. Abstr, Int. B 38 (1978) (10), 4676. MATHUR,1. H., and KEENAN, T. W.: Stability of alkaline phosphatase in sodium dodecylsulphate. FEES Let. 44 (1974) (1), 79-82. NAKAMURA, K., OKAZAWA, Y., SOEJIMA, M., and MASAKI, T.: Lysis of Micrococcus radiodurans. Agr. BioI. Chem. 3 (1973). NYBERG, H.: The effects of some detergents on the phosphatases and phosphate accumulation of Nitzschia actiruistroides, Ann. Bot. Fenn. 16 (1979) (1), 28-34. ONIYANAGI, T" FUG]SJiIMA, T., CCHIDA, K., and YOSHINO, H.: 5' Phosphodiesterase. Japan Kokai 77, 136, 989 (Cl. Cl. 2 D 13/10) 16.11. 1977, Appl. 76/52, 228, 10.5. 1976. 6 pp. PALAMARCZYK, G., LEHLE, L., and TANNER, W.: Polyprenylphosphate prevents inactivation of yeast glycotransferase by detergents. FEBS. Lett. 108 (1979) (1), 111-115. RUSSEL, R. R. B.: Use of Triton X-I00 to overcome the inhibition of fructosyltransferase by S.D.S. Anal. Biochem. 97 (1979) (I), 173-175. SHiI, E., and SIM, R. B.: Hydrodynamic parameters of the detergent solubilized hydrogenase from Pnracoccue denitr(ficans. Eur. J. Biochem. 97 (1979) (1), 119 -126. STARKOVA, Z., and STARKA, J.: Morphological mutants of E8cherichia coli: Nature of the permeability barrier in mon and env C cells. Ann. Microbiol. (Paris) 129 A (1978) (3), 265-284. SUGIRA, M., IsOBE, M.: Japan Kokai 7796, 790 (Cl. C 07 G 7)/02) 13th Aug. 1977. Appl. 76/11, 846, 07 Feb. 1976 4 pp. SUSMAN, M. L., and HAYS, J. B.: Irreversible inactivation of membrane-bound enzyme in II lac of the lactose phosphotransferase system of Staphylococcus aureus by Triton X-I00 and protection by substrates. Biochim. Biophys. Acta 465 (1977) (3), 559-570. TAMORI, Y., NISHIJIMO, M., and NOJIMA, S.: Properties of purified detergent-resistant phospholipase A of Escherichia coli K-I2. Inactivation and protection with detergents and phospholipids. TRIPATHI, V. S., and TRIPATHI, P.: Detergent effect on metabolic changes in microorganisms. A Review. Zbl. Bakt. II 135 (1980), 510-514. VAITKIRCIUS, R., CIPENS, L., MASIULYTE, G., and ANTONEKOVA, L. lVI.: Separation of intracellular amino peptidase from Bacillus eubtilie, Ortkrytiya, Izobret., Prom. Obraztsy, Tovarnye Znaki 55 (1978) (125), 102. VVILLETS, A. J.: Microbial metabolism of alkyl-benzene sulphonates. Fungal metabolism of 1phenylundecane-p-sulphonate and I-phenyldodecane-p-sulphonate. Antonie von Leeuwenhoek 39 (1976) (4), 567 -585. YOBE, S., MIROSE, Y., NAKAMURA, Y., and MITSUJI, K.: Alkaline proteinase production by bacterial fermentation. Ger. Offen-2, 313, 546 (C!. C. 12. d - CO 7 g), 27 Sept. 1973. Japan Appl. 72/27 G 73, March 18, 1972 , 15 pp. YUAN-CHI Su and SHu-YANU HWANG: Alkaline protease produced by Bacillus species. J. chin. Biochem. Soc. 2 (1973) (1), 1-10. Author's address: VIJAY SHANKAR TRIPATHI, Chemical Laboratories, Allahabad University, Allahabad, India.