Isolation of propioxatin A from Actinosynnema sp. SI-23 during a screening for Serratia piscatorum metalloproteinase inhibitors.

An inhibitor of Serratia metalloproteinase was found in the cultured broth of strain SI-23, which was identified to be Actinosynnema sp. The inhibitor had a potent inhibitory effect on Serratia metalloproteinase and a weak effect on thermolysin, but no effect on other groups of proteinases. The structure of the inhibitor was deduced to be propioxatin A, reported as an enkephalinase B inhibitor.

[Specific inhibitory effect of hybrid liposomes on the growth of hybridoma cells in vitro].

Remarkably high inhibitory effects of the hybrid liposomes composed of L-alpha-dimyristoyl-phosphatidylcholine (DMPC) and polyoxyethylenealkyl ether (C14(EO)n, n = 6-8 and C12(EO)n, n = 8-12)) on the growth of human lymphoma-human B-lymphocyte hybridoma (HF) cells in vitro were obtained. The hybrid liposomes composed of 90 mol% DMPC/10 mol% C14(EO)n (n = 6-8) or C12(EO)n (n = 8-12) were more fluid as compared with 90 mol% DMPC/10 mol% C14(EO)4 or C12(EO)n (n = 4, 23) hybrid liposomes on the basis of fluorescence polarization measurements. These results suggest that the inhibitory effects of the hybrid liposomes on the growth of HF cells should be related to the membrane fluidity.

Simple detection of xylosidase activity in single colonies, using 1-naphthyl-beta-D-xylopyranoside.

Since the xylosidase of Bacillus pumilus hydrolyzed 1-naphthyl-beta-D-xylopyranoside (naphthyl-X) to produce xylose and 1-naphthol and a chromogenic azo compound is produced by coupling 1-naphthol and Fast Blue Salt B, a simple method for detection of xylosidase activity in single colonies was studied. Escherichia coli JM109 carrying the xylosidase gene of B. pumilus was cultivated at 37 degrees C for 18 h on a LB plate containing 0.

A simple assay for xylanase using o-nitrophenyl-beta-D-xylobioside.

We measured xylanase activities upon eight chromogenic substrates, o- or p-nitrophenyl-beta-D-xylopyranoside (oNP-X or pNP-X) and o- or p-nitrophenyl-beta-D-xylooligosaccharides (oNP-Xn or pNP-Xn, n = 2-4), and studied for their uses as substrates in a kinetic study. The Kcat and K(m) of Bacillus pumilus xylanase (EC 3.2.

Purification and some properties of an endo-1,4-beta-D-mannanase from a marine mollusc, Littorina brevicula.

An endo-1,4-beta-D-mannanase (EC 3.2.1.

The terminal protein of the linear DNA plasmid pGKL2 shares an N-terminal domain of the plasmid-encoded DNA polymerase.

The 36K protein attached at the 5' end of the linear DNA plasmid pGKL2 from the yeast Kluyveromyces lactis was first purified and characterized. The terminal protein was purified from cells (1 kg wet weight) by ammonium sulphate precipitation and two rounds of centrifugation to equilibrium in CsCl gradients. The pGKL2 was present only in the post-microsomal supernatant.

Cytochrome P450rm from Rhodotorula minuta catalyzes 4-hydroxylation of benzoate.

Rhodotorula minuta, a red yeast, produces a cytochrome P450, tentatively named P450rm, catalyzing the formation of isobutene from isovalerate. We found that P450rm interacted with benzoate and the dissociation constant of P450rm for benzoate was 36 microM. A reconstituted system that consisted of purified P450rm and cytochrome P450 reductase catalyzed the 4-hydroxylation of benzoate in addition to the formation of isobutene; the turnover rate was approximately 40 nmol/min/nmol P450rm.

Improved Purification and Spectroscopic Properties of Squash Glutamate Decarboxylase.

Squash glutamate decarboxylase was purified by DEAE-Cellulose batchwise followed by Blue-Sepharose, Cellulofine GCL-2000, and Toyopearl HW-55F column chromatography. The purified glutamate decarboxylase had a high specific activity (95.0 u/mg).

Specific hybrid liposomes composed of phosphatidylcholine and polyoxyethylenealkyl ether with markedly enhanced inhibitory effects on the growth of tumor cells in vitro.

The hybrid liposomes (90mol% DMPC/10mol% C12(EO)8 and 90mol% DMPC/10mol% C12(EO)12) have a highly inhibitory action against the growth of tumor cells. The uniform and stable structure of the hybrid liposomes was revealed on the basis of electron microscopy and dynamic light scattering measurements.

Isolation and characterization of a novel 5-oxoprolinase (without ATP-hydrolyzing) from Alcaligenes faecalis N-38A.

A screening test was undertaken to isolate a microorganism that produced 5-oxoprolinase (without ATP-hydrolyzing). The 5-oxoprolinase (without ATP-hydrolyzing) activity (decyclization activity toward L-pyroglutamate) was found in a cell-free extract of Alcaligenes faecalis N-38A, newly isolated from a soil sample. The enzyme was purified as a homogeneous preparation.

Three ATP1 genes are present on chromosome II in Saccharomyces cerevisiae.

Chromosome fragmentation, ATP1 disruption, and Southern blot analyses of total DNAs and prime clones of chromosome II showed that three identical ATP1s are present, directing from the telomere to the centromere on the 35-55 kb far from the left telomere sequence of chromosome II. That is, the coding and 5'-, 3'-non-coding regions of ATP1 are repeated 3 times at approximately 7 kb intervals. These three ATP1s are expressed, and one and two ATP1s-disrupted strains, respectively, showed ca.

Migration of the yeast linear DNA plasmid from the cytoplasm into the nucleus in Saccharomyces cerevisiae.

The Kluyveromyces linear plasmids, pGKL1 and pGKL2, carrying terminal protein (TP), are located in the cytoplasm and have a unique gene expression system with the plasmid-specific promoter element termed UCS, which functions only in the cytoplasm. In this study we have developed an in vivo assay system in Saccharomyces cerevisiae which enables the detection of a rare migration of the yeast cytoplasmic plasmid to the nucleus, using a pGKL1-derived cytoplasmic linear plasmid pCLU1. pCLU1 had both the UCS-fused LEU2 gene (a cytoplasmic marker) and the native URA3 gene (a nuclear marker) and therefore its cytoplasmic-nucleo localized could be determined by the phenotypic analysis of the marker.

[Studies on the crude drugs used for the folk medicine described in "mimi-bukuro"].

"Mimi-bukuro" is a book written by Moriyasu Negishi in the Edo period. M. Negishi (1737-1815) was a magistrate in the town of Edo.

Increasing glutaminase activity in shoyu koji using a mixed culture of two koji moulds.

For improved fermentation of shoyu (soy sauce), a useful koji-making system has been developed using a mixed tane-koji of two shoyu koji moulds, namely Aspergillus oryzae K2 (length of conidiophores about 350 μm) and the late-conidiation strain, A. oryzae HG (length of conidiophores about 2500 μm). The mixed culture of strains K2 and HG had about twice the glutaminase activity of the single-strain cultures.

ATP1 and ATP2, the F1F0-ATPase alpha and beta subunit genes of Saccharomyces cerevisiae, are respectively located on chromosomes II and X.

Southern blot analysis showed that ATP1 and ATP2 map on chromosomes II and X, respectively. Physical mapping of ATP1 and ATP2 by chromosome fragmentation showed that ATP1 is at the left end of chromosome II and ATP2 is at the right end of chromosome X. Both are located close to telomere sequences of each chromosome; ATP1 and ATP2 being approximately 30 kb and 85 kb from the respective telomeres.

UV hypersensitivity of yeast linear plasmids.

The Kluyveromyces linear plasmids pGKL1 and pGKL2, encoding killer activity, were efficiently cured by UV irradiation. This event was investigated in more detail by the use of the terminal protein (TP)-associated cytoplasmic linear plasmids, pJKL1 and pRKL2, with a selectable marker LEU2. This observation was compared with the UV effect on the nuclear plasmids pLS1 (telomere-associated linear form) and YCp121 (centromere-integrated circular form), indicating that the UV hypersensitivity was specific to the cytoplasmic plasmids.

Ethylene production by strains of the plant-pathogenic bacterium Pseudomonas syringae depends upon the presence of indigenous plasmids carrying homologous genes for the ethylene-forming enzyme.

The molecular characteristics of the ethylene-forming enzymes of strains of Pseudomonas syringae were tested. The ethylene-producing activities of the nine strains as measured in vivo and in vitro were similar, except for that of P. syringae pv.

High affinity of hybrid liposomes for normal human epidermal keratinocytes in vitro.

A high affinity of hybrid liposomes towards normal human epidermal keratinocytes was observed. The fluorescence micrograph showed that hybrid liposomes may be incorporated into the keratinocytes by fusion.

Isolation and characterization of a dipeptidyl aminopeptidase from Streptomyces sp. WM-23.

A screening test was undertaken to isolate microorganisms that produced dipeptidyl aminopeptidase. The hydrolytic activity toward alanyl-phenylalanine p-nitroanilide was found in a culture filtrate of a actinomyces strain (WM-23), newly isolated from a soil sample. The enzyme (WM-23 dipeptidyl aminopeptidase) was isolated from the culture filtrate as a homogeneous preparation.

Reconstitution of the isobutene-forming reaction catalyzed by cytochrome P450 and P450 reductase from Rhodotorula minuta: decarboxylation with the formation of isobutene.

An isobutene-forming activity was reconstituted with cytochrome P450rm and cytochrome P450 reductase purified from Rhodotorula minuta. The nonionic detergent. Emulgen 911, present in the preparation of purified P450rm, inhibited the reconstitution.

[Studies on the medicinal plants in the "Sambutsu-cho" of Merayama, belonging to Kuma province, Higo].

The Sambutsu-cho, a list of the natural products, of Merayama, belonging to Kuma Province, Higo was dedicated to the Tokugawa shogunate ca. 1735. This book did not contain so many names, but there were various names of plants, animals and minerals.

Purification and characterization of cytochrome P450 from an isobutene-forming microorganism, Rhodotorula minuta.

A cytochrome P450 was purified from microsomes of Rhodotorula minuta. The optical spectrum of the purified cytochrome was characteristic of a low-spin ferric heme protein. Isovalerate caused a type I spectral change in it.

Purification and some properties of endo-1,4-beta-D-mannanase from a mud snail, Pomacea insularus (de Ordigny).

Endo-1,4-beta-D-mannanase (1,4-beta-D-mannanohydrolase, EC 3.2.1.