Sixty-three compounds were isolated and identified from heated starch and from heated starch with glycine. Starch or starch with glycine was heated in an Erlenmeyer flask at 290°C under a nitrogen stream. The volatiles formed during heating were entrained on a porous polymer (Porapack Q). Residual charred materials in the flasks were extracted with methylene chloride. The oily dark brown materials recovered from both the Porapack Q traps and the methylene chloride extracts were subjected to gas chromatography and gas chromatography/mass spectrometry. The compounds identified include 3 hydrocarbons, 1 1 ketones, 3 aldehydes, 24 aromatic compounds, 10 furans and 12 nitrogen-containing compounds.Formation mechanismsof somemajor products are proposed.

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A study was made of the incorporation of methanol and bicarbonate into the cell constituents of denitrifying or aerobic methanol grown and autotrophic H₂-O₂-CO₂grown Hyphomicrobium sp. 53-49. Cells were incubated with [¹⁴C]methanol or [¹⁴C]bicarbonate, and the distribution of the radioactivity in the nonvolatile constituents of ethanol extracts of cells was examined. When denitrifying grown cells were incubated with [¹⁴C]methanol, the major part of the radioactivity was fixed to serine as the first stable compound. Aerobic methanol grown cells also fixed [¹⁴C]methanol mainly to serine. These results suggest that methanol grown cells assimilate methanol by the serine pathway. Whendenitrifying or aerobic methanol grown cells were incubated with [¹⁴C]bicarbonate, malate was mainly observed as a nonvolatile compound in the initial period of the incubation. Autotrophic grown cells also fixed the major part of [¹⁴C]bicarbonate to malate. In this case, phosphoglyceric acid was found in the phosphorylated compounds area.

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Changes in the levels of free and protein amino acids in the callus of Hiproly barley were studied during differentiation. Adventitious roots were formed in the callus after.90 days of cultivation on modified White's medium containing 1 μM indoleacetic acid (IAA) and 200μM adenine sulfate, and callus placed on Murashige-Skoog's mediumwithout plant hormones formed adventitious roots after 30 days of cultivation. During differentiation, protein amino acids in the callus decreased, then increased, without an appreciable compositional change in the protein amino acids. The amount of free amino acids in the callus increased with root formation. The major free amino acids during differentiation were glutamine, asparagine, alanine, and proline. Glutamine increased until roots were found in the callus after cultivation. Asparagine gradually increased during differentiation.

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A new gallotannin named kurigalin was isolated from the bark of chestnut tree. The chemical structure of kurigalin was established as 2, 5, 6-tri-0-galloyl-α, β-D-hamamelose (I) on the basis of enzymatic hydrolyses and spectral analyses.

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The effect of KCD-232, a new hypolipidemic agent with the structure of 4-(4'-chloro-<benzyloxy)benzyl nicotinate, on cholesterol (Ch) metabolism was studied in rats fed a methioninesupplemented low casein (with threonine imbalance) diet.
A methionine-supplemented 10%casein (imbalance) diet caused an increase in serum Ch when compared to a 10%casein diet, while the supplement ofmethionine to a 20%casein diet caused no further increase. In rats fed the imbalance diet, KCD-232reduced the serum Ch level. Measurement of Chabsorption using a dual-isotope serum ratio methodshowedneither an enhancementin the imbalance diet-fed rats nor a reduction in the drug-treated, imbalance diet-fed rats. However, radioactivities in the serum of orally given [³H]Ch and intravenously given [¹⁴C]Ch increased in the imbalance diet-fed rats and decreased in the KCD-232-treated rats. Hepatic sterol synthesis measured in vivo using [³H] water was unchanged in the imbalance diet-fed rats and increased in the drug-treated rats.
These results suggest that the clearance of Ch from the circulation system, which maybe impaired in rats fed the imbalance diet, is enhanced in the KCD-232-treated, imbalance diet-fed rats.

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The kinetic properties of galactose oxidase of Gibberella fujikuroi were investigated. The enzyme oxidized D-galactose, 2-substituted D-galactoses, 2-epimer D-talose, and D-galactosecontaining oligosaccharides, but not other saccharides. The enzyme was highly specific for molecular oxygen as an electron acceptor. The enzymegave a nonlinear Lineweaver-Burk plot when the oxidation rate at a variety of galactose concentrations was investigated. The data were well explained by the assumption that the enzyme has two binding sites for galactose. K₁ and K₂, which are the dissociation constants for the first and second bindings of galactose to the enzyme, were calculated to be 1.3mM and 42mM, respectively. The 'enzyme was mixedly inhibited by dgalacturonic acid.

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The enzymatic preparation of laminaribiose was investigated in this research. Whencurdlan was hydrolyzed with the β-l, 3-glucanase system of Streptomyces sp. K27-4, the hydrolyzate mainly consisted of glucose and laminaribiose in an approximate ratio of 1 : 1 by weight. Yeast strains selected in this study, effectively and selectively metabolized all of the glucose in the hydrolyzate, without any degradation of laminaribiose.
By successive treatment of curdlan with the glucanase system and glucose-metabolizable yeast, 31g of crystalline laminaribiose was obtained from 1OOg of curdlan.

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Some new 2-(N-substituted or N, N-disubstituted aminoacetamido)-4-p-nitrophenyl, 4-m-nitrophenyl and 4-(2', 5'-dimethoxy)phenyl thiazoles were synthesised. Their hydrochlorides were screened for local anaesthetic activity by Frog's sciatic plexus method and this activity compared with that of procaine hydrochloride. Among them, the 2-(N-substituted aminoacetamido)-4-m-nitrophenylthiazole hydrochlorides are the most potent: almost twice in terms of the onset of anaesthesia in comparison with procaine-HCl.

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To find the function of the mannanchains covalently attached to yeast repressible acid phosphatase, the N-glycosidic carbohydrate chains were removed by endo-β-N-acetylglucosaminidase H under native conditions. Almost all of the N-glycosidic mannan chains were cleaved off by the glycosidase. The deglycosylated enzyme was shown to be a dimer structure as is the native enzyme. The deglycosylated enzyme retained enzyme activity, the same Km, and the same circular dichroism spectra as the native enzyme. These results indicate that the carbohydrate chains are not essential for maintaining the active enzyme structure, but the deglycosylated enzyme was shown to be more sensitive to acidic pHand high temperature.

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The pullulanase gene (pul) of Klebsiella aerogenes was transferred in vivo to Escherichia coli by using RP4: : Mu cts. The pul gene was expressed in E. coli, although the level ofpullulanase activity in E. coli was lower than that in K. aerogenes, and the Pul⁺ transconjugants were relatively unstable in an unselective medium. Production of pullulanase, which is used to makemaltose from starch, was induced in E. coli by pullulan, waxy maize amylopectin, soluble starch and maltose. Whenthe transconjugant cells of E. coli were grown with pullulan or maltose, most pullulanase was produced intracellularly, whereas K. aerogenes produced pullulanase extracellularly. Retransfer of the pul_k gene from E. coli to K. aerogenes by conjugation resulted in an increase of the production of extracellular pullulanase.

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Strains with lac fused to each of the arylsulfatase (ats) and tyramine oxidase (tyn) operons in Escherichia coli were isolated. Synthesis of β-galactosidase in strains with tyn : :lac fusions was induced by tyramine, histamine, tryptamine, dopamine and octopamine, and the induction of the tyn operon was subject to catabolite and ammoniumrepressions. These repressions were relieved whenthe cells were grown with a poor carbon or nitrogen source. No arylsulfatase activity is detected in E. coli strains. Synthesis ofβ-galactosidase in strains with ats : : lac fusions was repressed by sulfur compounds. The repression was relieved by monoamine compounds, which induced tyramine oxidase synthesis. The inhibition of tyramine oxidase activity by cysteine resulted in a decrease of the derepressed synthesis of β-galactosidase in the ats : : lac fusion. Repressing and derepressing conditions for the tyn operon prevented and stimulated, respectively, expression of the ats operon. Thus, the expression of latent arylsulfatase in E. coli seems to be regulated by expression of the tyn operon.

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An antifungal isoflavone, luteone [5, 7, 2′, 4′-tetrahydroxy-6-(3, 3-diniethylallyl)isoflavone] is metabolised by cultures of Aspergillus flavus and Botrytis cinerea into 2″, 3″-dihydro-3″-hydroxyluteone (AF- 1), 2″, 3″-dihydrodihydroxyluteone, a dihydrofuranoisoflavone (BG-1) and a dihydropyranoisoflavone. The structures of the metabolites were elucidated by physico-chemical and chemical procedures. The major metabolites, AF-1 and BC-1 are much less toxic than luteone against Cladosporium herbarum. The possible metabolic pathways are briefly discussed.

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Changes of flavor components of Sergia lucens Hansen (Sakura-ebi) during fermentation were studied by headspace analysis and the reduced pressure steam distillation method. Thirty two and 37 components were identified in the headspace volatiles and steam distillation concentrate, respectively, by GCand GC-MS.The major flavor components in the raw material such as trimethylamine, acetaldehyde, 2-methylpropanal and butanol decreased during fermentation. Alkylpyrazines and almost all the alcohols except butanol increased with the period of fermentation. From the result of the GC-sniff analysis, the components which are believed to contribute to the characteristic flavor of the fermented product are deduced to be 2, 5-dimethylpyrazine, 2, 6-dimethylpyrazine, 2, 3-dimethylpyrazine, isobutanol, isoamyl alcohol and isooctanol.

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Glutenin was reduced with various concentrations of 2-mercaptoethanol (2-ME) at pH 6.0 and the liberation of subunits was observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The amount of sulfhydryl groups in protein (protein-SH) liberated by reduction increased steeply with the increase of 2-MEconcentration around 1 mM.But the increase of protein-SH declined at higher concentrations to give a plateau at 6 mM.The level of this plateau, which may correspond to the amount of reactive disulfide (SS) bonds, was 40%of the total SS bonds of glutenin. In order to investigate whether inter-polypeptide SS bonds are cleaved by the partial reduction, the liberation of subunit monomers was observed by SDS-PAGE.By the partial reduction, Band 1+1′ (MW 104Kd), Band 2 (MW 90 Kd), Band 3 (MW 81 Kd) and Band 7 (MW 35 Kd) appeared in the electrophoretic pattern even at low concentrations of 2-ME, indicating the liberation of these subunits. However, although all monomersof Band 7 subunits were almost liberated at low concentrations of 2-ME (0.5mM), Bands 1+1′, 2, and 3 did not show suffic ient amounts of monomerseven at 6 mM.Further, another group of band, which had molecular weights of 136 Kd, 132 Kd, and 1 10 Kd, were observed at low concentrations of2-ME but dissapeared at extremely high concentrations, indicating these bands must be oligomers of glutenin subunits. These observations from the SDS-PAGE patterns suggested that the reactivities of interpolypeptide SS bonds differ according to the kinds ofsubunits. Band 6 (MW42 Kd) appeared at the position of MW38 Kd by reduction with lower concentrations of2-ME, indicating the retention of intra-polypeptide SS bonds. Bands 1+1′ 2, 3, and 7 that always appeared at the same positions independent of the 2-MEconcentrations, were separated from partial reduced glutenin (0.5 mMof 2-ME) and their SHand SS determined, in order to knowwhether these subunits retain intrapo lypeptide SS bonds. While the amount of liberated SH was much less than 2 mol per mol protein, the amount of SS bands was near 2mol, indicating that the liberated subunits retain intrapolypeptide SS bonds.

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Urease was purified to a homogeneousstate from cell-free extracts of a strain of Brevibacterium ammoniagenes.The enzymewas found to have a molecular weight of 200, 000, to consist of three identical subunits with a molecular weight of 67, 000, and to have an isoelectric point at pH 4. 1. The amino acid profile was similar to that of jack bean urease. Methionine and leucine were shown to be the N- and C-terminal residues, respectively. The enzyme contained 1 atom of nickel per subunit. The enzyme was most active at about pH 7 and at 65°C, and was stable betweeen pH 7 and 10 (at 30°C for 5hr) and below 50°C (at pH 7.5 for 1Omin). Among the compounds examined, Hg²⁺, Cu²⁺, PCMB, and hydroxyurea inhibited the enzyme strongly. The Km value of enzyme for urea was found to vary from 18 to 72mMdepending on pH and the type of buffer used. It was proved that the enzymecan be used for the determination of urea by the reaction rate assay method.

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The biosynthetic origin of the amino acid moieties of enduracidin was investigated by feeding experiments with labeled compounds. Results of the incorporation and the distribution of radioactivity into the antibiotic revealed that glycine, L-serine, L-threonine, L-alanine, L-aspartic acid, L-ornithine and L-citrulline were incorporated into the corresponding amino acid moieties. Unique amino acids, enduracididine and its isomer with an imidazolidine ring, were derived from larginine, but not histidine. K₁ (4-hydroxyphenylglycine) and K₂ (3, 5-dichloro-K₁) moieties were derived from L-tyrosine. ³⁶Cl-Sodium chloride was incorporated into the antibiotic in the early stage of fermentation.

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Corynebacterium equi IFO 3730 was found to oxidize a wide variety of sec-oalcohols, including alkanols, substituted alkanols, alkenols and cyclic alcohols, in moderate to high yields. Among them, the sec-alcohols which have longer carbon chains were oxidized more smoothly than those with smaller numbers of carbon. Although both enantiomers of unsymmetrically disubstituted carbinols were oxidized, the S form of 2-dodecanol was converted to the corresponding ketone a little faster than the other enantiomer.

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D-Gluconate dehydrogenase catalyzing the oxidation ofD-gluconate to 2-keto-D-gluconate was solubilized with Triton X-100 from the membrane of Gluconobacter dioxyacetonicus IFO 3271 and purified to an almost homogeneousstate by chromatographies on DEAE-cellulose and CMToyopearl in the presence of 0.1% Triton X-100. The enzyme had three subunits with molecular weights of 64, 000, 45, 000 and 21, 000, and contained approximately 2mol of heme per mol of the enzyme. The prosthetic group of the dehydrogenase was found to be a flavin covalently bound to the enzyme protein. The substrate specificity of the purified enzyme was very strict for D-gluconate and the apparent Michaelis constant for D-gluconate was 2.2mM.The optimum pH and temperature of the purified enzyme were 6.0 and 40°C, respectively.

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In the initial stage of hydrolysis, exo-β-(1→3)-D-glucanase from Basidiomycetes sp. QM806 cleaved laminaran from Eisenia bicyclis with a pattern resembling an erdo-hydrolase. Five kinds of intermediate gluco-oligosaccharides were separated by a combination of gel filtration and HPLC. They were shown to be 3²-O-β-D-glucosyl-gentiobiose, 3²-O-β-D-gentiobiosyl-gentiobiose, 3³-O-β-D-glucosyl-gentiotriose, 3⁴-O-β-D-glucosyl-3²-O-gentiobiosyl-gentiobiose, and 3³-O-β-D-gentiobiosylgentiotriose by enzymic hydrolysis and methylation analysis as well as by ¹³C NMRspectroscopy. As a result, such kinds of β(1→3)-β-(1→6)-linked oligosaccharides could be accounted for in the initial cleavage, and they were hydrolyzed ultimately to glucose, gentiobiose, and gentiotriose. It suggests that a single (1→3)-linkage on a block of (1→6)-links show some resistance to attack by this enzyme.

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Simple and speedy purification of Aspergillus oryzae metallo-proteinase was performed using Talopeptin-aminohexyl-Sepharose. The properties of the metallo-proteinase were: optimum pH 6.5; pH stability, pH 5-11; optimum temperature, 50°C; and molecular weight 42, 000 (SDS electrophoresis). These results were similar to those of neutral protease I from Aspergillus oryzae reported by Nakadai et al. This metallo-proteinase was compared with others from microbes using the metallo-proteinase inhibitors FMPI, PLT, and Talopeptin. The metallo-proteinase is unique in the point at which FMPI and PLT gave nearly stoichiometrical inhibition.

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Both ovalbumin and s-ovalbumin gave maximumgel strength at both sides of the isoelectric point. Maximumgel forming pHs of s-ovalbumin were almost the same as those of ovalbumin, but maximumgel strength values of s-ovalbumin were much smaller than those of ovalbumin. Although the gel strength of both proteins increased with increased heating temperature, the gel strength of s-ovalbumin was much smaller than that of ovalbumin at every heating temperature. About the results of creep experiments, all heat-induced gels were analyzed as a four-element model and the magnitude of all the parameters of both s-ovalbumin and intermediate was smaller than that of ovalbumin. Scanning electron microscopic studies showedthat the structure of ovalbumin gels was very fine comparing those of s-ovalbumin and the intermediate.

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Twelve bacterial strains which were concerned with dechlorination of 1, 2, 4-trichlorobenzene (TCB) were isolated from the intestinal contents of rats and it was found that they belonged to Staphylococcus epidermidis (strain A-F), Staphylococcus saprophyticus (strain G), Streptococcus sp. (strains H and I), Bacillus sp. (strain J), Gram negative rod (strain K) and Lactobacillus sp. (strain L).
In Staphylococcus epidermidis (Strain A), TCB was mainly converted to o-dichlorobenzene
and the latter was preferentially converted to monochlorobenzene (MCB) among dichlorobenzenes (DCBs). These conversions proceeded only under a gas phase of hydrogen. Furthermore, dry and broken cells of intact bacteria also maintained the dechlorinating activities, which were stimulated by the addition of NADPH.
Therefore, it was supposed that the conversion of TCB to MCB via DCBs was reductively carried out by enzymes originating from the isolated bacteria.

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Methionine-insensitive revertants with normal homoserine dehydrogenase (HD) derived from Brevibacterium flayum mutant No. 1-231, a lysine producer with S-(2-aminoethyl)-L-cysteine (AEC) resistance, methionine sensitivity, a low HDlevel and a pyruvate kinase (PK) defect, were still AEC-resistant and PK-deficient similar to No. 1-231. But they did not produce more lysine than the original strain, No. 15-8, from which strain No. 1-231 was derived. A high lysine producing mutant, No. 22, which was derived from strain No. 1-231, selected by sensitivity to β-fluoropyruvate (FP), and was defective in HD, produced more lysine than HD-defective mutants which were derived by two-step mutation from strain No. 1-231, selected by homoserine auxotrophy. Strain No. 22 did not showFP sensitivity under the conditions tested. Amongvarious lysine-biosynthetic enzymes examined, it had a higher level of aspartate-β-semialdehyde dehydrogenase than did its parent and the latter HD-defective mutants. Strain No. 22 produced 50 g/liter of lysine as the HC1 salt when cultured for 72 hr in a mediumcontaining soybean-meal hydrolysate, methionine and 100g/liter of glucose.

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The influence of 16 flavonoids on yeast α-glucosidase and β-fructosidase was investigated. Either p-nitrophenyl-α-D-glucopyranoside or maltose was used as substrate for α-glucosidase, and sucrose for β-fructosidase. Quercetin, myricetin, fisetin, and quercitrin strongly inhibited α-glucosidase at 0.1 mM. The concentrations which gave 50% inhibition were 4, 8, 8, 20, and 20 μM for myricetin, quercetin, fisetin, kaempferol, and quercitrin, respectively. The mode of inhibition was found to be mixed type, close to non-competitive type with quercetin and quercitrin. Fisetin, naringenin, and morin showed mixed type inhibition. Albumin from bovine serum slightly affected the anti-a-glucosidase activity of quercetin. The flavonoids tested exhibited little effect on β-fructosidase activity at 0.1 mM.

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L-Tartaric and L-malic acids in grape must were remarkably decomposed on incubation of Botrytis cinerea IFO 5964 under the conditions of 15 to 25% sugar content and pH 2.5 to 3.5 at 15 to 27°C. MoreL-tartaric acid was degraded by the fungus than L-malic acid in the absence of glucose.
Accumulation of ten organic acids was found in the mediumcontaining L-tartaric acid on incubation of B. cinerea. The main metabolites from L-tartaric acid with intact mycelia in 1/15 m phosphate buffer (pH 3.5 and 7.0) were L-lactic, acetic, D-glyceric, succinic, citric, L-malic, pyruvic and oxaloacetic acids.
The improvement of tartrate stability and sensory evaluation was achieved in the wine made from Riesling grape must which was blended with the botrytized must.

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The structure of soybean β-amylase in trigonal (P3₂21) crystals was determined at 4.5 A resolution by X-ray crystallographic techniques using the isomorphous replacement method. X Ray diffraction data were collected by the screened precession method for the native enzymeand two heavy atom derivatives. The shape of the enzymemolecule and the locations of mercurial binding are presented. The molecule appeared to be composed of two domains: the larger domain contains one mercurial site on its surface and the smaller domainhas another mercurial site, which seemed to be the so-called essential sulfhydryl group. A distinct cleft formed between the domains near the latter sulfhydryl group maybe a substrate binding region.

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Comparison of the effects of dietary PCB(0.03%) and caffeine (0.3%) on serum and liver lipids, and urinary ascorbic acid was done after different times. Serum total cholesterol, liver total lipids and triglyceride (TG) were found to continuously increase at 2, 4, and 8 weeks, but urinary ascorbic acid, serum TG and liver phospholipids were elevated up to 4 weeks in the PCB-fed rats. Liver cholesterol showed a decreasing trend after 2 weeks. On the other hand, dietary caffeine continuously increased serum cholesterol up to 8 weeks. Urinary ascorbic acid remained the same throughout the experimental period, but was significantly higher than in the respective controls at all times. SerumTGalso remained the same, but was lower than in the respective controls. Liver total lipids, cholesterol and TGdid not change in the caffeine-fed animals. The results clearly indicate that dietary PCBincreased all the parameters investigated whereas caffeine elevated serum cholesterol and urinary ascorbic acid, but depressed the serum TG concentration.

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To develop a rapid and convenient purification procedure for high molecular weight urokinase (the native form, mol wt., 54, 000), a reverse-immunoadsorbent which binds only contaminating proteins in the crude urokinase preparation was synthesized. Essentially all the impurity proteins were removed by adsorption to the anti-contaminant-IgG Sepharose 4B without conversion of the high molecular weight urokinase to the low molecular weight urokinase; three to four-fold purification of the enzyme was achieved by this single procedure with a yield of about 90%. The homogeneous high molecular weight urokinase was obtained easily by a following elution from a CM-Sephadexcolumn. The overall purification of the enzyme took less than three days with a final yield of 88%. The ability of the reverse-immunoadsorbent to adsorb impurity proteins was not reduced after six batch reactions. Reverse-immunoadsorption may be a generally useful method for protein purification.

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An amino acid detection system was developed for low picomole analysis of amino acids including proline, based on OPA-post-labeling with a non-switching NaCIO flow system. A computer-controlled HPLCequipped with a three-solvent gradient mixer was assembled with the detection system to analyze 17 kinds of amino acids in 85 min with a stable base line. The optimum conditions of the NaCIOand OPAsolutions were determined as 0.002%-0.22ml/min and 0.16%-0.26ml/min, respectively. The use of NaCIOcaused only approximately 30% decrease in fluorescence response of the usual amino acids, except for Pro and Cys, the latter even being enhanced by about 10 times. The detection limit for Pro and Gys was 500 and 1000 fmol, respectively, and that of the usual amino acids was 100 fmol. The calibration curves of all amino acids showed good linearity in a range between 5 and 500 pmol. This system was used as a detector for enantiomeric analysis of glutamic acid and cysteic acid with a reversed phase HPLC during stereochemical studies on the natural peptide derivative, oxycadystin.

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Glycerol dehydrogenase prepared from Cellulomonas sp. NT3060 by an improved purification procedure was characterized. The molecular weight wfas calculated to be about 390, 000 by gel filtration and about 336, 000 by the sedimentation equilibrium method. The enzymewas composed of eight identical subunits whose molecular weight was 42, 000-43, 000. The NH₂-terminal and COOH-terminal amino acids of the subunit were identified as serine and histidine, respectively. The octameric subunit model of the enzyme was confirmed by electron micrographs, which showed as octad aggregate, composed of two tetragons face to face. Studies on the initial velocity and product inhibition were consistent with an ordered Bi-Bi mechanism in which NAD⁺ is bound first to the enzyme and NADHreleased last.

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Extracellular glucoamylase produced by a starch-fermenting yeast, Saccharomyces diastaticus 5106-9A, was purified. The enzyme was found to be heterogeneous in molecular weight, ranging from approximately 80K to 66K as estimated by gel filtration, and consisted of two subunits, H and Y. The molecular weight of subunit Hwas heterogeneous and was determined to be approximately 68K, 59K, and 53K by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The molecular weight of subunit Y was 14K, estimated by the same gel. the molecular weight of the deglycosylated form of subunit H was 4IK, suggesting that the heterogeneity of the enzyme was due to glycosyl moieties of subunit H. Subunits H and Y were separated by gel filtration in the presence of sodium dodecyl sulfate. Subunit Y seemed to be hydrophobic, since it was insoluble in an aqueous buffer without detergent.

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A variety of benzimidazoles with a terpene chain and related compounds were assayed using the Tribolium cell-free chitin synthetase preparation. Amongthe compounds tested, 1-geranyland 1-nerylbenzimidazoles were the most potent inhibitors of in vitro chitin synthetase, and yet 12 times less inhibitory than polyoxin-D. The benzimidazoles gave steep slopes for the inhibition curves compared with polyoxin-D.

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Poly(A⁺) RNAwas isolated from lactating cow mammary gland and cDNAwas synthesized by a method which resulted in a high yield of products containing complete 5'-terminal mRNAsequences. Transformants were screened with a HaelU-EcoRI fragment for bovine α_s1-casein CDNA[M. Maki, M. Nagao, M. Hirose and H. Chiba, Agric. Biol Chem., 47, 441 (1983)]. A CDNAclone was isolated that contained the full length of α_s1-casein mRNAand its nucleotide sequence was determined. The amino acid sequence predicted from the nucleotide sequence was in good agreement with that determined with the purified mature α_s1-casein. The signal peptide consisting of 15 amino acids was identical to that of ovine α_s1-casein. High homology at the nucleotide sequence level in the signal peptide region and in the 5'-untranslated region was found between rat α-casein and bovine α_s1-casein.

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