The specificities and configurations of ternary complexes of yeast and liver alcohol dehydrogenases

1. Some aspects of the substrate specificities of liver and yeast alcohol dehydrogenases have been investigated with pentan-3-ol, heptan-4-ol, (+)-butan-2-ol, (+/-)-butan-2-ol, (+/-)-hexan-3-ol and (+/-)-octan-2-ol as potential substrates. The liver enzyme is active with all substrates tested, including both isomers of each optically active alcohol. In contrast, the yeast enzyme is completely inactive towards those secondary alcohols where both alkyl groups are larger than methyl and active with only the (+)-isomers of butan-2-ol and octan-2-ol. 2. The absence of stereospecificity of liver alcohol dehydrogenase towards optically active secondary alcohols and its broad specificity towards secondary alcohols in general are explained in terms of an alkyl-binding site that will react with a variety of alkyl groups and the ability of the enzyme to accommodate a fairly large unbound alkyl group in an active enzyme-NAD(+)-secondary alcohol ternary complex. The absolute optical specificity of the yeast enzyme towards n-alkylmethyl carbinols and its unreactivity towards pentan-3-ol, hexan-3-ol and heptan-4-ol are explained by its inability to accept alkyl groups larger than methyl in the unbound position in a viable ternary complex. 3. Comparison of the known configurations of the n-alkylmethyl carbinols and [1-(2)H]ethanol and [1-(3)H]geraniol, which have been used in stereospecificity studies with these enzymes by other workers, provides strong evidence for which alkyl group of the substrate is bound to the enzyme in the oxidation of n-alkylmethyl carbinols. The conclusions reached are, for butan-2-ol oxidation with the liver enzyme, confirmed by deductions from kinetic data obtained with (+)-butan-2-ol and a sample of butan-2-ol containing 66% of (-)-butan-2-ol. 4. Initial-rate parameters for the oxidations of (+)-butan-2-ol, 66% (-)-butan-2-ol and pentan-3-ol by NAD with liver alcohol dehydrogenase are presented. The data are completely consistent with a general mechanism of catalysis previously proposed for this enzyme.     

茶尺蠖绒茧蜂对茶梢挥发物的EAG和行为反应

为筛选有效诱集茶尺蠖绒茧蜂Apanteles sp. 的信息化合物及其组合, 选用了源于健康和虫害茶梢的27种典型挥发物的10-2 g / mL石蜡溶液、混合物1（含等量反-2-己烯醛、顺-3-己烯-1-醇和芳樟醇石蜡溶液）和混合物2（含等量反-2-己烯醛、顺-3-己烯-1-醇、2-戊烯-1-醇、反-2-戊烯醛、顺-3-己烯乙酸酯、正戊醇、正己醇和1-戊烯-3-醇石蜡溶液）, 用1~2日龄雌蜂为试虫, 测试其EAG反应, 并采用Y形嗅觉仪测定其行为反应; 另外, 选择5个茶园进行了野外生测试验。 EAG结果表明: 各味源的EAG值之间差异显著; 脂肪酸衍生物引起较强EAG反应, 其次为芳香化合物和异硫氰酸酯, 再次为倍半萜类和单萜类; 单组分中, 顺-3-己烯乙酸酯、反-2-己烯醛、水杨酸甲酯、反-2-戊烯醛、苯乙酮、苯乙醇、苯甲醇、苯甲醛和茉莉酸甲酯引起的EAG值较大, 1-戊烯-3-醇、2-戊烯-1-醇、顺-3-己烯-1-醇、香叶醇、罗勒烯、α-萜品烯、(+)-雪松醇、(+)-3-蒈烯、α-忽布烯和β-紫罗酮引起的值较小, Z-茉莉酮引起的最小; 混合物1引起的EAG值最大, 混合物2引起的较小. 使用EAG值较大的水杨酸甲酯、反-2-戊烯醛和混合物1等8种味源, 以Y形嗅觉仪进行的行为测定结果与EAG反应基本一致. 以正己烷为溶剂的10^-3, 10^-2和10^-1 g / mL水杨酸甲酯、10^-2 g / mL水杨酸甲酯和反-2-己烯醛混合溶液分别制成诱集剂, 载于橡皮头诱芯, 在浙滇闽粤茶园强烈地诱集茶尺蠖绒茧蜂、单白绵绒茧蜂和其他茧蜂, 并表现梯度效应。据此认为虫害诱发的茶梢互利素引起该蜂强烈EAG反应和趋向性, 互利素与互利素或普通植物挥发物的恰当组合可于茶园中有效诱集该蜂。     

Comparison of the olfactory sensitivity of two sympatric steppe grasshopper species (Orthoptera: Acrididae) to plant volatile compounds

Two Oedipodinae grasshopper species Oedaleus decorus asiaticus B.-Bienko and Angaracris baraben-sis Pall. (Orthoptera: Acrididae) are important pests on the natural grasslands in Inner Mongolia and often require insecticide treatment during outbreaks[1]. They both prefer overgrazed steppes and xerophytous habi-tats, and have thus been suggested as indicator species for steppe deterioration in typical steppe zones of In-ner Mongolia[2]. The two species have a sympatric distribution and sync…     

Microbial hydroxylation of unsaturated, cyclic carboxylic acids protected as benzoxazoles

Microbial hydroxylation of 2-(cyclopent-1-enyl)benzoxazole (1) and 2-(cyclohex-1-enyl)benzoxazole (2) by Cunninghamella blakesleeana DSM 1906 and Bacillus megaterium DSM 32, respectively, gave chiral allylic alcohols 3-(benz-1,3-oxazol-2-yl)cyclopent-2-en-1-ol (3) and 3-(benz-1,3-oxazol-2-yl)cyclohex-2-en-1-ol (4) along with achiral ketones 3-(benz-1,3-oxazol-2-yl)cyclopent-2-en-1-one (5) and 3-(benz-1,3-oxazol-2-yl)cyclohex-2-en-1-one (6). Both allylic alcohols were produced in enantiomeric excesses higher than 99%. The determination of their absolute configurations (S in both cases) is described.     

Syntheses of alpha-D-galactosamine neoglycolipids

Several N-acetyl-alpha-d-galactosamine neoglycolipids, as well as hydrophobized T and T(N) antigen analogues, were prepared for embedment onto liposomes. Three different lipidic structures were used for the anchoring, that is cholesterol, 1,3-bis(undecyloxy)propan-2-ol and 1,3-bis(3,7,11,15-tetramethylhexadecyloxy)propan-2-ol. Oligoethyleneglycol spacers were used to link the carbohydrate and the hydrophobic moieties; their lengths were varied in order to obtain model compounds for the selective recognition by sialyl transferases involved in cancer processes. Glycosylation reactions were optimized to sluggish amphiphilic acceptor alcohols, in order to reach good 1,2-cis-stereoselectivities and acceptable yields. This aim was achieved by using 3,4,6-tri-O-acetyl-2-azido-2-deoxy-d-galactopyranosyl trichloroacetimidate as the donor, trimethylsilyl trifluoromethanesulfonate as the promoter and diethyl ether or mixtures of diethyl ether and dichloromethane as solvents.     

HS-SPME-GC/MS analysis of the volatile compounds of <Emphasis Type="Italic">Achillea collina</Emphasis>: Evaluation of the emissions fingerprint induced by <Emphasis Type="Italic">Myzus persicae</Emphasis> infestation

A Headspace Solid-phase Microextraction (HS-SPME) method combined with Gas Chromatography-Mass Spectrometry (GC/MS) was developed and optimized to extrat and analyze the volatile compounds of aerial parts of Achillea collina Becker ex Rchb. and to investigate the effect of the phlem feeding aphid Myzus persicae Sulzer on the Volatile Organic Compounds (VOCs) emitted by the infested plants. The extraction of 1 g of powdered freeze dried plant samples for 120 min at 30°C using divinylbenzene-carbowax-polydimethylsiloxane (DVB/CAR/PDMS) fiber showed the highest area counts for the majority of the volatile compounds. Overall, 62 and 80 volatile compounds were detected in control and infested plant samples respectively. In A. collina infested plants, we observed a great increase in both monoterpenes and sesquiterpenes fractions. Several changes among alcohols also occurred, particularly regarding Z-3-hexen-1-ol, E-3-hexen-1-ol and E-2-hexen-1-ol proposing these compounds as herbivore-induces plant volatiles (HIPVs). New perspective for agricultural practice may derive from the opportunity to identify novel herbivores-induced plant VOCs active as plant protection agents.     

Purification and characterization of a NAD+-dependent secondary alcohol dehydrogenase from propane-grown Rhodococcus rhodochrous PNKb1

NAD⁺-linked primary and secondary alcohol dehydrogenase activity was detected in cell-free extracts of propane-grown Rhodococcus rhodochrous PNKb1. One enzyme was purified to homogeneity using a two-step procedure involving DEAE-cellulose and NAD-agarose chromatography and this exhibited both primary and secondary NAD⁺-linked alcohol dehydrogenase activity. The Mr of the enzyme was approximately 86,000 with subunits of Mr 42,000. The enzyme exhibited broad substrate specificity, oxidizing a range of short-chain primary and secondary alcohols (C₂–C₈) and representative cyclic and aromatic alcohols. The pH optimum was 10. At pH 6.5, in the presence of NADH, the enzyme catalysed the reduction of ketones to alcohols. The K _m values for propan-1-ol, propan-2-ol and NAD were 12 mM, 18 mM and 0.057 mM respectively. The enzyme was inhibited by metal-complexing agents and iodoacetate. The properties of this enzyme were compared with similar enzymes in the current literature, and were found to be significantly different from those thus far described. It is likely that this enzyme plays a major role in the assimilation of propane by R. rhodochrous PNKb1.Abbreviations HPLC high performance liquid chromatography - DEAE diethyl amino ethyl - IEF isoelectrofocusing - NTG nitrosoguanidine - SDS-PAGE sodium dodecylsulphate polyacrylamide gel electrophoresis - pI isoelectric point     

Pine shoot beetle, Tomicus piniperda (Col., Scolytidae), responses to common green leaf volatiles

We tested the hypothesis that green leaf volatiles (GLVs) disrupt the response of overwintered pine shoot beetles, Tomicus piniperda (L.) to multiple-funnel traps baited with the attractive host volatile α-pinene. A combination of four GLV alcohols, 1-hexanol ( E )-2-hexen-1-ol ( Z )-2-hexen-1-ol, and ( Z )-3-hexen-1-ol, caused 54 and 36% reduction in the number of pine shoot beetles captured in two separate trapping experiments. Similarly, a combination of the four alcohols plus two GLV aldehydes, hexanal and ( E )-2-hexenal, caused 38% reduction in the number of pine shoot beetles captured compared with α-pinene alone. A blend of the two GLV aldehydes was not disruptive. None of the four GLV alcohols nor the two GLV aldehydes were disruptive when tested individually. The finding that the blend of four GLV alcohols reduced attraction of T. piniperda supports the general hypothesis that GLVs common to nonhost angiosperms are disruptive to conifer-attacking bark beetles (Scolytidae).     

Dialkylmethyl beta-glycosides of N-acetylmuramyl-L-alanyl-D-isoglutamine: synthesis and infection protective and cytotoxic activities

Symmetric secondary linear alcohols were proposed as aglycones for the synthesis of lipophilic beta-glycosides of N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP). Pentadecan-8-ol, nonadecan-10-ol, and tricosan-12-ol were glycosylated by the oxazoline method. Based on the corresponding glucosaminides, alkyl beta-glycosides of 4,6-O-isopropylidene-N-acetylmuramic acid were synthesized and coupled with the dipeptide. Deprotection of isopropylidene groups by acidic hydrolysis and catalytic hydrogenolysis of benzyl esters resulted in the target muramyldipeptide glycosides. Nonadecan-10-yl and tricosan-12-yl [beta]-MDPs at doses 2 microg/mice most effectively stimulated antibacterial resistance in mice against Staphylococcus aureus. In contrast to the previously synthesized undecan-6-yl beta-MDP, pentadecan-8-yl, nonadecan-10-yl, and tricosan-12-yl beta-MDPs demonstrated direct cytotoxicity toward tumor cells E-562 and blood mononuclear cells.     

The preparation and application of functionalised synthetic oligonucleotides: III. Use of H-phosphonate derivatives of protected amino-hexanol and mercapto-propanol or -hexanol.

Syntheses of H-phosphonate salts (4a-e) of N/S-protected alcohols such as 6-aminohexan-1-ol, 3-mercaptopropan-1-ol and 6-mercaptohexan-1-ol are described using 2-chloro-5,6-benzo-1,3,2-phosphorin-4-one (2) as the phosphonylating agent. The H-phosphonate salts (4a-e), in the presence of pivaloyl chloride or adamantoyl chloride as an activator, were coupled to the 5'-end of synthetic oligonucleotides on solid supports to produce amino or thio-linked oligonucleotides. Following deprotection and purification, fluorescent dyes, biotin derivatives and poly-L-lysine-maleimide were separately attached to the functionalised oligonucleotides. Identical derivatized oligomers were obtained with cyanoethyl-N,N-diisopropylamidite chemistry and amidites (5a-e) of the respective alcohols.     

Synthesis of silicon-containing azole derivatives with magnesium bromide diethyl etherate,and an investigation of their fungicidal activities

Enhancement of magnesium bromide diethyl etherate in addition reaction of trimethylsilylmethylmagnesium chloride 5 to 2-(1H-1,2,4-triazol-1-yl)acetophenones 6 allowed us to prepare silicon-containing azole derivatives, which were tested for fungicidal activity and phytotoxicity. Among them, 2-(4-fluorophenyl)-1-(1H-1,2,4-triazol-1-yl)-3-trimethylsilylpropan-2-ol 1a was determined to be the most effective and potential candidate for novel fungicide.     

Effect of ageing on sterol metabolism in potato tuber slices

When fresh potato tuber slices were incubated with [1-¹⁴C]-sodium acetate, cycloartenol was heavily labelled but no radioactivity was recovered in 24-methylene cycloartanol and free sterols. If potato slices were aged for 0–24 hr before feeding with radioactive acetate, a rapid increase of the label in the sterol precursors and the free sterols was observed. The free sterol content was 5 × higher after ageing for 24 hr. Isofucosterol synthesis was especially stimulated. The synthesis of sterols during the ageing process seems to be related to the appearance of a cycloartenol C24-methylase and may be linked to a biogenesis of membranes.Nomenclature: (1) 4,4,14α-trimethyl 9β, 19β-cyclo-5α-cholest-24-en 3β-ol; (2) 4,4,14α-trimethyl 9β, 19β-cyclo-5α-ergost-24(28)-en 3β-ol; (3) 4α,14α-dimethyl 9β,19β-cyclo 5α-ergost 24(28)-en 3β-ol; (4) 4α, 14α-dimethyl 5α-ergosta 8.24(28)-dien 3β-ol; (5) 4α-methyl 5α-ergosta 7,24(28)-dien 3β-ol; (6) ergosta 5,24(28)-dien 3β-ol; (7) stigmasta 5,Z-24(28)-dien 3β-ol; (8) (24R)-24 methyl cholest 5-en 3β-ol; (9) (24R)-24 ethyl cholest 5-en 3β-ol; (10) (24S)-24 ethyl cholesta 5,E-22(23)-dien 3β-ol; (11) cholest 5-en 3β-ol.     

Novel very long-chain methyl-branched alcohols and their esters,and methyl-branched alkanes in pupae of the cabbage looper,Trichoplusia ni (Hubner)

Four homologous series of very long-chain methyl-branched alcohols were found in the internal lipids of cabbage looper, Trichoplusia ni, pupae both as free alcohols and as esters. These alcohols were identified based on mass spectra of their TMS and acetate derivatives, and on the Kovats Indices and mass spectra of the alkanes obatained by reduction of their bromide derivatives with LiAlH₄. The four homologous series (from C36 to C46) consisted of monomethyl-, two dimethyl- and a trimethyl-branched alcohol series. The major alcohols (with the corresponding alkanes in parentheses) were identified as 24-methyltetracontan-1-ol (17-methyltetracontane), 24,28-dimethyltetracontan-1-ol (13,17-dimethyltetracontane), 24,36-dimethyltetracontan-1-ol (5,17-dimethyltetracontane) and 24,28,36-trimethyltetracontan-1-ol (5,13,17-trimethyltetracontane). The minor components had an odd number of carbon atoms (C37, C39, C41 and C43) in the carbon chain and the methylalkanes formed from their reduction had methyl branches on the 18- and 14,18-positions. The methylalkanes found internally in pupae were similar to those previously reported in larval cuticular lipids (de Renobales and Blomquist, Insect Biochem. 13, 493–502, 1983). Additional novel methylalkanes identified were 2,X-dimethylalkanes, 5,15-dimethylhentriacontane, 5,23-dimethyltritriacontane, 5,17,23-trimethyltritriacontane and 5,15,23-trimethylpentatriacontane. The methyl branching positions of the major methyl-branched alcohols were different from those of the major methylalkanes.     

Volatile compounds of endophyte-free and infected tall fescue (Festuca arundinacea Schreb.).

Volatile compounds produced by intact plants and ground leaf tissue from endophyte-infected (E+) and endophyte-free (E-) tall fescue (Festuca arundinacea Schreb.) were collected by a purge-and-trap procedure and analyzed by gas chromatography/mass spectrometry The volatile compound profile from ground leaf tissue was similar between E+ and E- clonal plants; however, the sheaths of E+ clonal plants produced higher levels of 1-octen-3-ol, a characteristic volatile compound derived from lipid peroxidation in fungi, which was absent in E- clonal plants. Intact plants produced fewer volatiles than macerated leaves. At 25 degrees C, (Z)-3-hexen-1-ol acetate was the most abundant compound, accounting for 77 and 89% of the total volatile emission from E+ and E- plants, respectively. Higher temperature (32 degrees C) significantly reduced the production of (Z)-3-hexen-1-ol acetate. Nonanal was the most abundant compound at 32 degrees C accounting for 52 and 45% of the total volatile emission from E+ and E- plants. Treatment of E+ and E- plants with jasmonic acid (JA) dramatically altered the volatile compound profile. The levels of (E)-beta-ocimene increased more than 200-fold and accounted for at least 43% of the total volatile emission. Although the presence of endophyte resulted in some qualitative and quantitative differences in the production of volatile compounds, they are unlikely to account for the differences in insect resistance between E+ and E- plants. Nevertheless, the production of a unique spectrum of volatiles after JA treatment may represent a significant plant-based defense response in tall fescue that is independent of endophyte.     

Practical enantioresolution of alcohols with 2-methoxy-2-(1-naphthyl)propionic acid and determination of their absolute configurations by the (1)H NMR anisotropy method.

The enantioresolution of racemic alcohols as esters of 2-methoxy-2-(1-naphthyl)propionic acid (MalphaNP acid 1) and the determination of their absolute configurations on the basis of (1)H NMR anisotropy effect are described. The enantiopure MalphaNP acid (S)-(+)-1 was allowed to react with racemic 2-alkanols and 1-octyn-3-ol, yielding diastereomeric mixtures of esters, which were easily separated by HPLC on silica gel. To determine the absolute configurations of the first-eluted diastereomeric esters by the (1)H NMR anisotropy method, the general scheme was proposed. Separated esters were reduced with LiAlH(4) or hydrolyzed with KOH/EtOH to recover enantiopure alcohols.     

Comparison of the olfactory sensitivity of two sympatric steppe grasshopper species (Orthoptera: Acrididae) to plant volatile compounds

Electroantennogram (EAG) responses of male and female Oedipodinae grasshoppers, Oedaleus decorus asiaticus B.-Bienko and Angaracris barabensis Pall to 37 plant volatile compounds were recorded. The two species have sympatric distribution and synchronous seasonal activity in Inner Mongolia Grasslands. They have different host plant preference with Oedaleus decorus asiaticus graminivorous and A. barabensis forbivorous. The EAG response profiles to 37 compounds of the two species and their both sexes were similar. Most of the green leaf volatiles elicited higher EAG responses in both species and sexes than terpenic compounds and some aromatic compounds. Strong EAG responses were elicited by 6-carbon alcohols (1-hexanol, Z-hexen-2-ol-1, E-2-hexen-1-ol, E-hexen-3-ol-1), aldehyde (E-2-hexen-1-al), ester (Z-hexen-3-yl acetate), and 7-carbon alcohols (1-heptanol) in both species and sexes. Monoterpenes with functional groups of alcohols and aldehydes were more stimulating than other monoterpenes tested. The sesquiterpenes tested elicited relatively low responses. Benzaldehyde elicited the highest responses for both species among aromatic components. Oedaleus decorus asiaticus showed higher EAG responses than A. barabensis to the green leaf volatiles, 1-decanol, 1-nonanol, 1-pentanol, hexanal, Z-hexen-3-yl acetate and to the sesquiterpenes (-)-trans-caryophyllene. Males have higher responses than females in Oedaleus decorus asiaticus. No sexual difference was observed in A. barabensis. Dose-dependent responses to six selected chemicals were observed from females. In both species, all the chemicals tested elicit EAG responses at concentration between 10^-3 mol/L and 10^-2 mol/L, except that the responses of A. barabensis to terpineol had a threshold concentration of 10-2 mol/L benzaldehyde and 1-hexanol had the highest slopes in dose curves, while 1-octen-3-ol showed the smallest slope in responses to the six chemicals tested. Comparative studies on the responses of two antennal sections and the whole antenna to six selected chemicals were carried out using females of both species. A significant EAG response was only recorded from the distal part of the antenna and not from the proximal seven segments.     

Synthesis of Short-Chain Diols and Unsaturated Alcohols from Secondary Alcohol Substrates by the Rieske Nonheme Mononuclear Iron Oxygenase MdpJ

The Rieske nonheme mononuclear iron oxygenase MdpJ of the fuel oxygenate-degrading bacterial strain Aquincola tertiaricarbonis L108 has been described to attack short-chain tertiary alcohols via hydroxylation and desaturation reactions. Here, we demonstrate that also short-chain secondary alcohols can be transformed by MdpJ. Wild-type cells of strain L108 converted 2-propanol and 2-butanol to 1,2-propanediol and 3-buten-2-ol, respectively, whereas an mdpJ knockout mutant did not show such activity. In addition, wild-type cells converted 3-methyl-2-butanol and 3-pentanol to the corresponding desaturation products 3-methyl-3-buten-2-ol and 1-penten-3-ol, respectively. The enzymatic hydroxylation of 2-propanol resulted in an enantiomeric excess of about 70% for the (R)-enantiomer, indicating that this reaction was favored. Likewise, desaturation of (R)-2-butanol to 3-buten-2-ol was about 2.3-fold faster than conversion of the (S)-enantiomer. The biotechnological potential of MdpJ for the synthesis of enantiopure short-chain alcohols and diols as building block chemicals is discussed.     

NAD(+)-dependent (S)-specific secondary alcohol dehydrogenase involved in stereoinversion of 3-pentyn-2-ol catalyzed by Nocardia fusca AKU 2123

An NAD(+)-dependent alcohol dehydrogenase was purified to homogeneity from Nocardia fusca AKU 2123. The enzyme catalyzed (S)-specific oxidation of 3-pentyn-2-ol (PYOH), i.e., part of the stereoinversion reaction for the production of (R)-PYOH, which is a valuable chiral building block for pharmaceuticals, from the racemate. The enzyme used a broad variety of secondary alcohols including alkyl alcohols, alkenyl alcohols, acetylenic alcohols, and aromatic alcohols as substrates. The oxidation was (S)-isomer specific in every case. The K(m) and Vmax for (S)-PYOH and (S)-2-hexanol oxidation were 1.6 mM and 53 mumol/min/mg, and 0.33 mM and 130 mumol/min/mg, respectively. The enzyme also catalyzed stereoselective reduction of carbonyl compounds. (S)-2-Hexanol and ethyl (R)-4-chloro-3-hydroxybutanoate in high optical purity were produced from 2-hexanone and ethyl 4-chloro-3-oxobutanoate by the purified enzyme, respectively. The K(m) and Vmax for 2-hexanone reduction were 2.5 mM and 260 mumol/min/mg. The enzyme has a relative molecular mass of 150,000 and consists of four identical subunits. The NH2-terminal amino acid sequence of the enzyme shows similarity with those of the carbonyl reductase from Rhodococcus erythropolis and phenylacetaldehyde reductase from Corynebacterium sp.     

Biosynthesis of secondary alcohols and ketones from alkanes

Combined gas-liquid chromatography-mass spectrometry of the secondary alcohols and ketones from broccoli (Brassica oleracea) showed that the former consisted of a mixture of nonacosan-14-ol (43%) and nonacosan-15-ol (57%) whereas the latter contained predominantly nonacosan-15-one (92%). Chemical degradation of the secondary alcohols derived from [4-¹⁴C]stearic acid in B. oleracea indicated that the intact C₁₈ chain was incorporated into nonacosan-14-ol and nonacosan-15-ol and that the hydroxyl groups of the positional isomers were introduced into a preformed aliphatic chain. [G-³H]Nonacosane was incorporated into nonacosan-14-ol and nonacosan-15-ol in B. oleracea in a ratio similar to that found in the natural mixture. A randomly tritiated natural mixture of secondary alcohols was incorporated into the ketone fraction by broccoli leaves. Chemical degradation of substrate secondary alcohols and the ketones biologically derived from them demonstrated that nonacosan-15-ol was the preferred secondary alcohol for biological oxidation to the ketone. The incorporation of [G-³H]nonacosane into nonacosanol and nonacosanone by broccoli leaves required O₂ and was inhibited by phenanthroline. The inhibition was reversed by Fe²⁺ suggesting that conversion of nonacosane into the oxygenated derivatives involve a mixed-function oxidase. From these results it is concluded that in B. oleracea nonacosane is hydroxylated to give nonacosan-15-ol and nonacosan-14-ol with subsequent preferential oxidation of the C-15 isomer to give the ketones.     

Olfactory discrimination of structurally similar alcohols by cockroaches

The capability of the cockroach Periplaneta americana to discriminate odors of structurally similar aliphatic alcohols was studied by using an operant conditioning paradigm. Cockroaches were trained to discriminate three odors: one odor associated with sucrose solution (reward) and two odors associated with NaCl solution (non-reward). After training, their odor preferences were tested by counting the number of visits to each odor source. We tested the capability of cockroaches to discriminate (1) three normal aliphatic alcohols with different numbers of carbon (1-pentanol, 1-hexanol and 1-octanol), (2) three C6 aliphatic alcohols (1-hexanol, 2-hexanol and trans-2-hexen-1-ol), (3) binary mixtures of two of these three alcohols and their components, and (4) 1-hexanol solution of three different concentrations (1, 10 and 100 micro g micro l(-1)). Cockroaches exhibited higher preferences for the odors associated with reward in these tests, and we therefore conclude that cockroaches can discriminate these odors. However, discrimination of 1-hexanol and trans-2-hexen-1-ol and their binary mixture was imperfect, in that some statistical tests suggested significant level of discrimination but other tests did not. In addition, the cockroaches learned to associate a 1-hexanol solution of the highest or lowest concentration with sucrose reward but failed to learn to associate 1-hexanol of an intermediate concentration with reward.