All issues
Volume 66 (1969)
- Issue 6 Pages 751-
- Issue 5 Pages 573-
- Issue 4 Pages 431-
- Issue 3 Pages 289-
- Issue 2 Pages 123-
- Issue 1 Pages 11-
Volume 66, Issue 4
Displaying 1-17 of 17 articles from this issue
- |<
- <
- 1
- >
- >|
-
II. Study on the Specificity of NepenthesinSHIZUKO AMAGASE, SHIGERU NAKAYAMA, AKIRA TSUGITA1969 Volume 66 Issue 4 Pages 431-439
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe substrate specificity of the partially purified acid protease in Nepenthes (nepenthesin) was studied by hydrolyzing several peptides of known structures. The results indicated that nepenthesin was predominantly specific to aspartic acid residue at its carboxyl side or at its amino side, and was apparently specific also to tyrosine and alanine residues at their carboxyl sides. The protease seemed to be an endopeptidase.View full abstractDownload PDF (409K) -
HARUKI YAMAGUCHI, TOMOHIRO MEGA, TOKUJI IKENAKA, YOSHIO MATSUSHIMA1969 Volume 66 Issue 4 Pages 441-444
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. α- and β-Mannosidases [EC 3. 2. 1. 24, EC 3. 2. 1. 25] and N-acetyl-β-glucos-aminidase [EC 3. 2. 1. 30] were used for the elucidation of the anomeric configuration of mannose and N-acetylglucosamine residues in the glycopeptide prepared from Taka-amylase A [EC 3. 2. 1. 1].
2. Most of the mannose linkages in the glycopeptide were found to be α-glycosidic, because α-mannosidase liberated all of the mannose residues of the glycopeptide.
3. It was shown by the action of N-acetyl-β-glucosaminidase on the degraded glycopeptide (D2-GP) that one N-acetyl glucosamine residue situated at the nonreducing end of D2-GP had a β-linkage.View full abstractDownload PDF (287K) -
SHUZO OTANI, TAKETO YAMANOI, YOSHITAKA SAITO1969 Volume 66 Issue 4 Pages 445-453
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe ornithine activating enzyme was purified about thirty-fold from Bacillus brevis NAGANO cells. This enzyme had a high specificity for ornithine and ornithine analogues failed to stimulate the PP1-ATP exchange reaction. Radioactive ornithine was not transferred to tRNA by this enzyme. Activity appeared in the late logarithmic phase of cell growth and increase in enzyme activity was coincident with gramicidin S formation. From these results, it seems likely that the ornithine activating enzyme is involved in gramicidin S formation.
Full enzyme activity was obtained on addition of magnesium ions, while sulfhydryl reagents were inhibitory.View full abstractDownload PDF (564K) -
MUTUMI MURAMATU, SETSURO FUJII1969 Volume 66 Issue 4 Pages 455-462
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe reaction of diazomium-1H-tetrazole (DHT)* with trypsin [EC 3. 4. 4. 4] caused complete inactivation of the enzyme. However, in the presence of a substrate, such as methyl Nα-tosyl-L-arginate, or hexyl δ-guanidinovalerate, which is a potent competitive inhibitor of trypsin, DHT caused 60 to 70 per cent loss of trypsin activity. Nα-Benzoyl-L-arginine, δ-guanidinovaleric acid, and hexyl γ-guanidinobutyrate have no effect on inactivation of trypsin by DHT.
The product of the reaction of trypsin with DHT in the presence of hexyl δ-guani-dinovalerate (active DHT-trypsin) has a spectrum with maxima at 320 to 322 mμ and at 500mμ. The absorption at 320mμ of active DHT-trypsin was greater than that of the reaction product of trypsin in the absence of inhibitor (inactive DHT-trypsin), and the absorption at 500mμ was less than that of inactive DHT-trypsin.
Amino acid analysis showed that in active DHT-trypsin, seven tyrosyl-, one histidyl- and thirteen lysyl residues are modified with DHT, and in inactive DHT-trypsin, eight tyrosyl-, two histidyl- and thirteen lysyl residues are modified.
The reaction of tetranitromethane (TNM)* with trypsin also reduced the enzyme activity. However, inactivation was prevented to a considerable extent by the presence of hexyl ε-aminocaproate, which is a potent competitive inhibitor. Amino acid analysis showed that seven and eight tyrosyl residues are nitrated in the presence of hexyl δ-aminocaproate and in its absence, respectively.View full abstractDownload PDF (568K) -
X. The “Denitrifying Enzyme” as a Nitrite Reductase and the Electron Donating System for DenitrificationMACHIKO MIYATA, TAKESHI MORI1969 Volume 66 Issue 4 Pages 463-471
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe “denitrifying enzyme”, a soluble copper protein obtained from a strain of Pseudomonas denitrificans, is designated as a nitrite reductase catalyzing the reduction of nitrite to nitric oxide with reduced Pseudomonas cytochrome c-553 or cytochrome c-552 as the electron donor, from the following results.
1. Using lactate and yeast lactate dehydrogenase [EC 1. 1. 2. 3] as the electron donating system, hydrogen-, or electron transport took place through bacterial cytochrome c-552 or cytochrome c-553 (1) to the “denitrifying enzyme” reducing nitrite. Ps. stutzeri cytochrome c-552, 558 (2) or oxidation-reduction dyes such as thionine, brilliant cresyl blue and methylene blue could replace Ps. denitrificans cytochromes. With each of these electron donating systems nitrite was converted only to nitric oxide.
2. In the ascorbate-TMPD-nitrite system, cytochrome c-553 was a better electron carrier than cytochrome c-552 for the “denitrifying enzyme”, producing nitric oxide. Ps. stutzeri cytochrome c-552, 558 could replace cytochrome c-553.
3. The “denitrifying enzyme” catalyzed nitrite reduction, oxygen consumption in the presence of ascorbate, TMPD and cytochrome c-553 and hydroxylamine oxidation in the presence of nitrite. However, it was more active with nitrite than with oxygen or hydroxylamine.
Addition of flavin nucleotides enhanced both the rate of DCIP reduction by the bacterial lactate dehydrogenase, which appeared to be a particulate bound enzyme, and the rate of denitrification of the particulate fraction plus soluble fraction or cell-free extract. However, flavins did not increase the NO reducing activity of the particulate fraction or stimulate nitrite reduction by the soluble fraction alone. These facts suggest that flavin nucleotides act in electron transport from the particulate system to the soluble system.
Nicotinamide nucleotides appeared to have no essential role in the lactate-nitrite system.View full abstractDownload PDF (594K) -
TATSUHIKO YAGI1969 Volume 66 Issue 4 Pages 473-478
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSFormate dehydrogenase and its electron acceptor were isolated and purified from Desulfovibrio vulgaris.
The optimum pH of the enzyme (K3Fe (CN)6 as an acceptor) is 6-7. Km for formate is 7.4×10-6M. NAD+, NADP+, FAD, FMN, cytochrome c3 and clostridial ferredoxin are not reduced by the enzyme in the presence of formate. The enzyme is not inhibited by p-chloromercuribenzoate, iodoacetamide, sodium hypophosphite, EDTA or o-phenanthroline, but strongly inhibited by KCN.
The natural electron acceptor is a c-type cytochrome. The absorption spectrum of its reduced form has α, β and γ peaks at 553, 523 and 416 mμ, respectively. Its molecular weight is about 6500.
Desulfovibrio formate dehydrogenase is, thus, formate: ferricytochrome c 553 (mol. wt., 6500) oxidoreductase, a new entry in EC class 1. 2. 2.View full abstractDownload PDF (390K) -
I. The Incorporation of 32P-Orthophosphate into PhospholipidsMASAYUKI KATAYAMA, SABURO FUNAHASHI1969 Volume 66 Issue 4 Pages 479-485
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSwollen shelled seeds or etiolated seedlings after three days' germination of mung bean, Phaseolus radiatus var. typicus were incubated with 32P-orthophosphate solution at 30°C in the dark. The 32P incorporation into phospholipids in the cotyledons of the former, or in the hypocotyls and radicles of the latter was observed by time course study in terms of their water-soluble deacylation products. The greater part of the incorporated radioactivity was observed in glycerophosphorylcholine (GPC) from the cotyledons, and in glycerophosphorylethanolamine (GPE) from the hypocotyls and radicles. Specific radioactivity was given as follows in decreasing order: Glycerophosphorylserine (GPS), GPC>GPE>glyeerophosphorylinositol (GPI) in the cotyledons, and GPS>GPE>GPC in the hypocotyls and radicles. The specific radioactivity of GPI fraction was remarkable in the hypocotyls and radicles. The different patterns of phospholipid metabolism observed between the cotyledons and the hypocotyls and radicles were discussed in connection with biosynthetic pathways of phosphatidylcholine.View full abstractDownload PDF (464K) -
I. Properties of an Adenosine-sensitive MutantMAKOTO KIDA, FUMIKO KAWASHIMA, AKIRA IMADA, IKUO NOGAMI, IKUO SUHARA, ...1969 Volume 66 Issue 4 Pages 487-492
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAn adenosine-sensitive mutant was derived from a strain of Bacillus subtilis. The growth of the mutant was strongly inhibited by the presence of adenosine at 0.1mM and the inhibition was completely reversed by the addition of guanine derivatives, but not by other purine and pyrimidine derivatives. The growth of this mutant was also inhibited by psicofuranine (9-D-psicofuranosyl-6-aminopurine), which is a structural analog of adenosine and is known to suppress GMP synthesis by inhibiting XMP aminase [EC 6. 3. 4. 1, XMP: ammonia ligase (AMP)].
Adenosine-resistant mutants were derived from the sensitive mutant. XMP aminase was partially purified from the adenosine-sensitive and resistant strains as well as the parent strain. The activity of XMP aminase from the sensitive strain was strongly inhibited by either adenosine or psicofuranine, while the enzymes from the resistant and parent strains were little affected by both inhibitors. Thus the adenosine sensitivity of the mutant may be attributed to the inhibition of its XMP aminase by adenosine.View full abstractDownload PDF (393K) -
YUKIO IKEHARA, SUSUMU YANAGI, HIDEYA ENDO1969 Volume 66 Issue 4 Pages 493-501
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSMuscle aldolases [EC 4. 1. 2. 13] have been purified from rat, rabbit, human, pigeon, and hen. No significant differences among the five enzymes were detected in electrophoresis, ultracentrifugal analysis and kinetic properties. But some differences were observed in immunochemical investigation by means of double diffusion analysis and enzyme inhibition in the presence of antibodies against rat, human or pigeon muscle aldolase. Fowl anti-rat (or anti-human) muscle aldolase sera cross-reacted completely with other two mammalian aldolases, but did little with aldolases isolated from avians (hen and pigeon). Guinea pig anti-pigeon aldolase sera cross-reacted with hen aldolase, but did not with those of mammalians. It is concluded that in immunochemical and biochemical properties, three mammalian muscle aldolases are remarkably similar with each other, but those of birds lack some of the antigenic determinants present in the mammalian enzymes; even between hen and pigeon, the former aldolase lacks some of the determinants in that of the latter.View full abstractDownload PDF (2920K) -
YOSHIKO KASHIWABARA, HIROSHI SUZUKI, KAZUTOSI NISIZAWA1969 Volume 66 Issue 4 Pages 503-512
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSTwo kinds of pseudomonads, Nos. 8 and 9 were isolated from sea sand and decaying algal fronds. Crude extracts from these bacterial cells grown on an alginate medium and the partially purified enzyme preparations obtained therefrom degraded eliminatively various substrates such as commercial alginate, mannuronic acid-rich (M-rich) and guluronic acid-rich (G-rich) alginates, a short-chain polyguluronide with -DP 15 (SG) and its reduced derivative and a short-chain polymannuronide with -DP 11 (SM) in different manners. However, a highly purified lyase preparation from Pseudomonas No. 9 showed the activity for SG but not for SM, and the independent existence of a “polyguluronide lyase” was ascertained.View full abstractDownload PDF (1406K) -
Studies on Circular DichroismKIYOSHI IKEDA, KOZO HAMAGUGHI1969 Volume 66 Issue 4 Pages 513-520
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe binding of N-acetyl-D-glucosamine to lysozyme [EC 3. 2. 1. 17] was studied by the circular dichroism (CD) technique. The increase in the CD band of lysozyme at 294 mμ resulting from binding of the inhibitor was utilized to study the interaction. It was found that the association was optimal at pH's 3.5 to 5.5 due to ionization of two groups with apparent pK's of about 2.5 and 6.5. The binding site of N-acetyl-glucosamine was discussed in relation to X-ray crystallographic data on lysozyme. The pH dependence of the CD spectrum in the absence of the inhibitor was also described.View full abstractDownload PDF (479K) -
VIII. Particle-bound L-Sorbose Dehydrogenase from Gluconobacter suboxydansKIYOSHI SATO, YUZO YAMADA, KÔ AIDA, TEIJIRO UEMURA1969 Volume 66 Issue 4 Pages 521-527
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAn L-sorbose-dehydrogenating enzyme was found in a particulate fraction of Gluconobacter suboxydans grown on a sorbitol medium. The reaction with 2, 6-dichlorophenol indophenol as acceptor was formulated as; L-sorbose+acceptor→5-keto-D-fructose+ reduced acceptor. The optimal pH for the activity was 6.4. Km value for L-sorbose was calculated as 4.3×lO-3M. p-Chloromercuribenzoate, phenylmercuric nitrate, Ag+, Hg2+, and Cu2+ inhibited the enzyme activity to considerable degrees. D-Fructose, sucrose, and dulcitol were inactive as substrate. The enzyme preparation had the activity toward aldoses, D-gluconate, 2-keto-n-gluconate, and polyols. It was, however, proved that the L-sorbose-dehydrogenating activity was due to a separate enzyme other than glucose dehydrogenase, gluconate dehydrogenase [EC 1. 1. 99. 3], ketogluconate dehydrogenase [EC 1. 1. 99. 4], and mannitol dehydrogenase [EC 1. 1. 2. 2]. Thus, this enzyme, to which the rule of Bertrand-Hudson is not applicable, was proposed as a new type of “ketose dehydrogenase”.View full abstractDownload PDF (489K) -
MICHIYUKI YAMADA, KIYOSHI KURAHASHI1969 Volume 66 Issue 4 Pages 529-540
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSPhenylalanine racemase of Bacillus brevis NAGANO which produces gramicidin S was purified to a nearly homogeneous state. The purification procedure involved ultracen-trifugation, ammonium sulfate fractionation, calcium phosphate gel adsorption, gel-filtration with Sephadex G-150, and chromatography on DEAE-Sephadex A-50. The nearly homogeneous enzyme preparation possessed two enzymatic activities; phenylalanine racemization and phenylalanine activation. The molecular weight of the enzyme was estimated to be 100, 000 by the sucrose density gradient centrifugation method. Activa-tion energy was estimated to be 10.9×103 cal per mole. The Km, values for L-phenyl-alanine and ATP were determined to be 2.0×10-5M and 1.5×10-4M, respectively. The velocity of the D-phenylalanine formation from the L-isomer was markedly affected by the concentration of dithiothreitol and pH, whereas that of L-phenylalanine formation from the n-isomer was less affected. ATP was consumed during the conversion of L-phenylalanine into the n-isomer with the concomitant formation of AMP. AMP as well as PP; enhanced the enzyme activity several fold.View full abstractDownload PDF (1360K) -
II. Purification of the Pancreatic ProteasesATSUSHI KOIDE, YOSHITAKA MATSUOKA1969 Volume 66 Issue 4 Pages 541-548
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe separation and purification of three kinds of proteolytic enzymes from fin whale pancreas are described.
An acid extract of fin whale pancreas was fractionated with (NH4)2SO4 and treated with trypsin [EC 3. 4. 4. 4]. An anionic chymotrypsin [EC 3. 4. 4. 5] and a cationic proteolytic enzyme fraction in this crude proteolytic enzyme preparation were roughly separated by CM-Sephadex column chromatography.
Cationic chymotrypsin and trypsin were obtained from the cationic proteolytic enzyme fraction by CM-cellulose column chromatography.
These proteolytic enzymes were purified further by rechromatography on either CM-Sephadex or CM-cellulose.View full abstractDownload PDF (2848K) -
HACHIRO NAKAGAWA, SHOICHI MIURA, HIROSHI KIMURA, TAKAHIRO KANATSUNA1969 Volume 66 Issue 4 Pages 549-564
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSerine dehydratase [EC 4. 2. 1. 13] was induced by feeding a synthetic diet containing L-serine as the sole source of non-essential amino nitrogen (Serine-diet). Of the non-essential amino acids tested, only L-serine caused induction, the extent of which was dependent on the serine content of the diet. From these experiments, it was concluded that induction of this enzyme by its substrate was based on the requirement of the nitrogen moiety for growth under the conditions employed.
It was demonstrated that the increase in enzyme activity resulting from this Serine-diet, was blocked by cycloheximide. By pulse-labeling with 14C-amino acids followed by specific precipitation of the enzyme protein with the antiserum, it was confirmed that the induction of serine dehydratase was due to increased net synthesis of enzyme protein.
It was also demonstrated that activation of serine dehydratase during increased gluconeogenesis, such as on treatment of adrenalectomized-diabetic rats with triamcinolone, was based on increased net synthesis of enzyme protein.
Evidence was presented, however, for a difference in the biological significances of substrate, and hormonal inductions of serine dehydratase. Under hormonal control, serine is converted to sugar via pyruvic acid and the resulting ammonia is metabolized via the urea cycle. Under substrate induction, on the other hand, the α-amino group is used in formation of other non-essential amino acids needed for growth and the residual carbon skeleton may be metabolized via the TCA cycle.View full abstractDownload PDF (1194K) -
KEIICHI YOSHIMURA, TSUTOMU HIROSHIGE, SHINJI ITOH1969 Volume 66 Issue 4 Pages 565-567
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (208K) -
MASACHIKA IRIE1969 Volume 66 Issue 4 Pages 569-572
Published: October 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (246K)
- |<
- <
- 1
- >
- >|