A novel lactonohydrolase, an enzyme that catalyzes the hydrolysis of aldonate lactones to the corresponding aldonic acids, was purified 10-fold to apparent homogeneity, with a 61 % overall recovery, from Fusarium oxysporum AKU 3702, through a purification procedure comprising DEAESephacel, octyl-Sepharose CL-4B and hydroxyapatite chromatographies and crystallization. The molecular mass of the native enzyme, as estimated by high-performance gel-permeation chromatography, is 125 kDa, and the subunit molecular mass is 60 kDa. The enzyme contains 15.4% (by mass) glucose equivalent of carbohydrate, and about 1 mol calcium/subunit. The enzyme hydrolyzes aldonate lactones, such as D-galactono-y-lactone and L-mannono-y-lactone, stereospecifically. Furthermore, it can catalyze the asymmetric hydrolysis of D-pantoyl lactone, which is a promising chiral building block for the chemical synthesis of D-pantothenate. These reactions are reversible, and the reaction equilibrium at pH 6.0 has a molar ratio of nearly 1 : 1 with D-pantoyl lactone and D-pantoic acid. The K, and V,,, for D-galactono-y-lactone are 3.6 mM and 1440 U/mg, respectively, and those for D-galactonate are 52.6 mM and 216 U/mg, respectively. The enzyme also irreversibly hydrolyzes several aromatic lactones, such as dihydrocoumarin and homogentisic-acid lactone.Lactonohydrolases (lactonases) reversibly catalyze the hydrolysis of lactone compounds into hydroxy acids, i.e. they mediate the interconversion between the lactone and acid forms of hydroxy carboxylic acids. Among them, gluconolactonase (aldonolactonase), L-arabinonolactonase and D-arabinonolactonase specifically hydrolyze aldonate lactones to their respective aldonic acids. Although the activities of these enzymes have been detected in various organisms [l -81, there has been no report of the purification of one of these enzymes to homogeneity, except for bovine liver aldonolactonase [5], and so ceI1-free extracts or partially purified preparations have been used for characterization of the enzymes in most cases. Therefore, there has not been sufficient study of the enzyme chemistry. On the other hand, as to the roles of these enzymes in vivo, Bulitz and Lehninger suggested that aldonolactonase participates in the formation of L-gulono-y-lactone from L-gulonate in L-ascorbate biosynthesis [3]. Recently, L-rhamnonolactonase and D-xylOnOlactonase were found in Pulluluriu pulluluns and Pseudomonas frugi, respectively, being involved in the oxidative degradation of L-rhamnose and D-xylose, respectively [9, 101.

We have previously reported induction of fatty acid synthase (FAS) gene expression by insulin and adipocyte differentiation in 3T3-L1 cells. In order to identify sequences responsible for insulin regulation of the FAS gene, chimaeric constructs containing serial deletions of the 5'-flanking region of the rat FAS gene ligated to the chloramphenicol acetyltransferase (CAT) reporter gene were prepared and transfected into 3T3-L1 cells. Plasmids containing 2100 (-2100CAT), 1400 (-1400CAT), 1009 (-1009CAT) and 332 (-332CAT) bp of FAS 5' flanking sequences exhibited comparable basal CAT activities in 3T3-L1 preadipocytes. This activity was 3-fold higher when these constructs were transiently transfected into 3T3-L1 adipocytes. Stably transfected 3T3-L1 cells also exhibited a 3-fold increase in CAT activity upon adipocyte differentiation, indicating that sequences required for the differentiation-dependent increase in FAS expression are located within the 332 bp promoter. Treatment with 10 nM insulin increased CAT activity by 2.1 +/- 0.2-, 2.6 +/- 0.1-, 2.0 +/- 0.2- and 1.7 +/- 0.2-fold respectively in 3T3-L1 adipocytes transiently transfected with -2100CAT, -1400CAT, -1009CAT and -332CAT plasmids. CAT activity was increased by 3.0 +/- 0.3- and 3.5 +/- 0.6-fold respectively by insulin treatment in adipocytes stably transfected with -2100CAT and -1009CAT plasmids. When insulin-responsive H4IIE hepatoma cells were transiently transfected with -2100CAT, -1400CAT, -1009CAT and -332CAT plasmids and then treated with 10 nM insulin, CAT activity increased by 3.1-, 3.1 +/- 0.8-, 3.0 +/- 0.7- and 2.3 +/- 0.5-fold respectively in serum-free media, and by 2.6 +/- 0.4-, 3.3 +/- 0.9-, 3.1 +/- 0.4- and 2.9 +/- 0.6-fold respectively in the presence of 0.5% serum. These results indicate that sequences responsible for insulin regulation of FAS gene are also located within 332 bp of the transcription start site.

Improvement of electrical properties for 7.4 × 10 17 cm −3 phosphorus-doped pulsed laser crystallized silicon films of 50 nm thickness formed on quartz glass substrates was achieved by heat treatment with high-pressure H 2 O vapor. The electrical conductivity was increased from 1.3 × 10 −5 S/cm (as-crystallized) to 2 S/cm by annealing at 270 • C for 3 h with 1.3 × 10 6 Pa H 2 O vapor. The spin density of undoped laser crystallized silicon films was reduced from 1.6 × 10 18 cm −3 (as-crystallized) to 1.2 × 10 17 cm −3 by annealing at 310 • C for 3 h with 1.3 × 10 6 Pa H 2 O vapor. Theoretical analysis revealed that the potential barrier height at grain boundaries decreased from 0.3 eV (as-crystallized) to 0.002 eV. High-pressure H 2 O vapor annealing offer the possibility of reducing the density of defects states through oxidation of the defects at low temperature.

The home garden "peharangan" in Indonesia is the traditional garden with agroforestry and sustainable land use in the tropical regions. The objective of the present study was to elucidate the effects of urbanization on the performance of the home gardens in West Java, Indonesia.The study sites were ranked into the least urbanized site, the intermediate urbanized site, and the most urbanized site. The performances of the home gardens were compared among these different urbanized sites. The home gardens in the most urbanized sites were derived from the dry land and mixed garden, and those in the least and intermediate urbanized sites had been the home gardens for along time, or were derived from mixed gardens or from rice fields. The area of home gardens tended to decrease due to urbanization, and the shape was restricted to block and strip pattern in the most urbanized sites. The frequencies of front yards were high in all the sites, and side yardstended to decrease due to urbanization, but the back yards were remaining even in the most urbanized sites. From the point of the composition of the elements in the home gardens, it was suggested that properties of the recycling system for agroforestry and the sociality or the public utilization were diminishing in the home gardens due to the urbanization.

Native DL-pantothenic acid, having a 1,3-diol structure, was chirally resolved by ligand exchange capillary electrophoresis using (S)-3-amino-1,2-propanediol as a chiral selector and the borate anion as a central ion. The optimum conditions for both high resolution and short migration time of DL-pantothenic acid were found to be 200 mM (S)-3-amino-1,2-propanediol and 200 mM borate buffer (pH 9.2) containing 15% methanol with an applied voltage of +25 kV at 20 degrees C, using direct detection at 200 nm. With this system, the resolution (Rs) of racemic pantothenic acid was approximately 1.7. When (S)-1,2-propanediol, (S)-1,2,3-propanetriol, (S)-1,3-butanediol or (S)-1-amino-2-propanol were used as chiral ligand instead of (S)-3-amino-1,2-propanediol, DL-pantothenic acid was not enantioseparated. When borate was replaced with Tris or butylborate, no chiral separation was achieved. Therefore, the ionic interaction between the amino and carboxyl groups of the ternary complex may play an important role in the enantioseparation of DL-pantothenic acid by the proposed CE system.