Lipase-catalyzed synthesis of butyl butyrate by alcoholysis in an integrated liquid-vapor system

This paper reports experimental work pertaining to alcoholysis between butanol and ethyl butanoate, catalyzed by an immobilized lipase in a liquid-vapor system where chemical reaction and physical separation are simultaneously carried out. The processing setup was tested for various compositions of the starting feedstock and operated under reduced pressure. Samples were withdrawn both from the boiler and the condenser, and they were chromatographically assayed for butyl butyrate. The integrated configuration tested is quite effective toward improvement of the final yield of the desired product.     

Kinetic model for synthesis of fructosyl-stevioside using suspended β-fructofuranosidase

The synthesis experiments of fructosyl-stevioside were conducted under the various conditions of the initial concentrations of the substrates and the enzyme. The transfructosylation of stevioside with sucrose and the hydrolyses of sucrose and fructosyl-stevioside simultaneously occurred. The fructosyl-stevioside synthesis was inhibited by the side products, glucose and fructose. A kinetic model was constructed by considering the Ping-Pong Bi Bi mechanism for the transfructosylation, the apparent Ordered Uni Bi mechanism for the hydrolysis and the competitive inhibition by the side products. The model constants were estimated by fitting the model equations with the experimental results for the sucrose hydrolysis and the fructosyl-stevioside synthesis. The model can predict not only the appropriate conditions to efficiently synthesize the fructosyl-stevioside, but also the reaction time giving the maximum conversion.     

Regioselective acylation of disaccharides in tert-butyl alcohol catalyzed by Candida antarctica lipase

The acylation of several disaccharides by ethyl butanoate and ethyl dodecanoate was catalyzed by Candida antarctica lipase in tert-butyl alcohol, at temperatures ranging from 40 degrees to 82 degrees C (reflux temperature). The relative reaction rates of the various disaccharides were directly related to their solubility. The primary products were the monoesters derived from acylation of the primary alcohol groups. At higher conversions diesters were formed, and the ratio of diester to monoester was markedly dependent on the structure of the disaccharide. Thus, reaction of maltose with ethyl dodecanoate in refluxing tert-butyl alcohol afforded the 6'-monododecanoate even at high conversions. Trehalose, in contrast, afforded the 6,6'-diester. Acylation of the less soluble sucrose and lactose was much slower, but a moderate (37%) conversion of sucrose was observed after a prolonged reaction time (7 days). A number of other lipases and proteases were tested but C. antarctica lipase was unique in catalyzing the acylation of sucrose in refluxing tert-butyl alcohol. (c) 1996 John Wiley & Sons, Inc.     

Synthesis of short chain alkyl esters using cutinase from Burkholderia cepacia NRRL B2320

Short chain alkyl esters are well appreciated for fruity flavors they provide. These are mainly applied to the fruit-flavored products like jam, jelly, beverages, wine and dairy. Cutinase from Burkholderia cepacia NRRL B 2320 was found to be active in catalyzing the synthesis of alkyl esters in organic solvent. The optimal temperature range for the enzyme catalyzed synthesis was found to be from 35 °C to 40 °C. The maximum conversion (%) during synthesis of ester was obtained for butyric acid (C4) and valeric acid (C5) with butanol reflecting the specificity of the enzyme for short-chain length fatty acids. In case of alcohol specificity, butanol was found to be most preferred substrate by the enzyme and conversion (%) decreased with increasing carbon chain length of alcohol used in the esterification reaction. The kinetic analysis for the synthesis of butyl butyrate by varying concentration of one substrate at a time (butanol or butyric acid), showed that Ping–Pong Bi Bi model with acid inhibition and influence of initial water is most suitable model for the prediction of the reaction kinetics.     

Random Bi Bi机制的非稳态酶动力学布尔函数图论研究

本文以非稳态酶动力学的布尔函数图形方法^[1],来研究一类Random Bi Bi机制的非稳态酶动力学问题,推导同此类反应的非稳态酶动力学方程,并对此动力学方程进行了讨论,分析了此类Random Bi Bi机制酶反应体系的非稳态酶动力学方程。     

Ping Pong Bi Bi机制的非稳态酶动力学分布函数图论研究

以非稳态酶动力学的布尔函数图形方法,来研究一类PingPongBiBi机制的非记酶动力学问题,推导出此类反应的非稳态酶动力学方程,并对此动力学方程进行了讨论,分析了此类PingPongBiBi机制酶反应体系的非稳态酶动力学方程。     

Kinetic cutinase-catalyzed esterification of caproic acid in organic solvent system

Practical application of any chemical reaction requires the knowledge of its kinetics; in particular if one wishes to be able to describe a chemical reactor over an extended range of reaction conditions or if one intends to optimize the reaction conditions, a suitable kinetic model must be obtained. In order to ensure that the model is applicable over a wide range of experimental conditions it should be based on a mechanistic scheme describing the fundamental steps involved in the reaction; the development of these kind of models can also be used to provide insight into the processes that are taking place.A kinetic study, using experiments carried out in a batch stirred reactor, has been made for the enzymatic esterification of caproic acid with ethyl alcohol catalyzed by Fusarium solani pisi cutinase. Different acid and alcohol concentrations (whilst also varying the acid/alcohol molar ratio) were tested and the results were used to identify the best reaction scheme to describe the results obtained over an extended range of conditions. Several different approaches were used to identify the most adequate mechanistic model, namely by resorting to the quasi stationary state and the rate-limiting hypothesis. The main kinetic characteristics observed in esterification reaction were found to follow an ordered Ping-Pong Bi–Bi mechanism but different modifications were used o ensure that the kinetic model was applicable over the entire range of experimental conditions that were covered.     

Effects of pear fruit-derived volatiles and their concentrations on the responses of oriental fruit moths

Grapholita molesta (Busck) is an important pest of pear trees. Numerous esters accumulate consistently in mature pear fruits. However, little is known about the effects of single esters from pear fruits at different concentrations on the responses of male and female G. molesta. In this study, the responses of virgin males, virgin females, and gravid females to five esters (3-methylbutyl acetate, ethyl hexanoate, ethyl butanoate, butyl acetate, and hexyl acetate) of pear fruits at three dosage levels (1, 5 and 10 μg/μL) were investigated using electroantennography (EAG), wind tunnel experiments, and field tests. Our results showed that ethyl butanoate (5 μg/μL) elicited higher EAG responses in virgin females and males than a mixture of all five volatiles, and elicited an equal EAG response to that for the mixture in gravid females, with all responses to ethyl butanoate and the mixture being higher than the response to hexane alone. For upwind flight, ethyl butanoate (5 μg/μL) excited equal moth flight–location behavior in virgin and gravid females to that for the mixture, which were both higher than the response to hexane. For close flight, ethyl hexanoate (5 μg/μL) and 3-methylbutyl acetate (10 μg/μL) appeared to be the activating compounds, eliciting equal responses from virgin females to that of the mixture, and higher responses of gravid females to that of the mixture, both of which were higher than the responses to hexane. Upwind and close flight responses of virgin males were lower than those of virgin and gravid females. In field trials, we verified that traps baited with ethyl butanoate (5 μg/μL) captured more moths than the other esters. Therefore, from the esters and concentrations tested, we recommend ethyl butanoate (5 μg/μL) for potential use as an attractant for G. molesta in orchards.     

Insect pest control agents: Novel chiral butanoate esters (juvenogens)

During the investigation of ester derivatives (juvenogens, biochemically activated insect hormonogenic compounds) of biologically active alcohols with potential application in insect pest control, a need for availability of all existing stereoisomers of ethyl N-{2-[4-(2-butanoyloxycyclohexyl)methyl]phenoxy}ethyl carbamate occurred. They were synthesized from their chiral precursors, the corresponding stereoisomers of 2-(4-methoxybenzyl)cyclohexyl butanoate, by removing their protecting group (methyl), and by subsequent condensation of the aromatic hydroxyl moiety with ethyl N-(2-bromoethyl) carbamate. The requested enantiomers of 2-(4-methoxybenzyl)cyclohexyl butanoate were obtained by a Candida antarctica lipase-mediated transesterification and chiral resolution of the respective racemic cis- and trans-isomers of 2-(4-methoxybenzyl)cyclohexanol either directly or after a subsequent chemical esterification of the chiral precursor. In this synthesis, two convenient butanoic acid activating esters, vinyl butanoate and 2,2,2-trifluoroethyl butanoate, were employed, and the chiral precursors in the synthesis of the target molecules were obtained in 41-48% yields (i.e., 82-96% conversion), and with enantiomeric purity ee=96-98%, respectively. The enantiomeric purity of the products was determined by chiral HPLC analysis, and their absolute configuration was assigned on the basis of analyzing the (1)H and (19)F NMR spectra of their diastereoisomeric Mosher acid (3,3,3-trifluoromethyl-2-methoxy-2-phenylpropanoic acid) esters.     

Evaluation of free radical scavenging activity of Butea monosperma Lam

In the present study, ethyl acetate, butanol and aqueous fractions derived from total methanol extract of Butea monosperma flowers were evaluated for radical scavenging activities using different in vitro models like reducing power assay, scavenging of 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, hydroxyl radical and inhibition of erythrocyte hemolysis using 2, 2' azo-bis (amidinopropane) dihydrochloride (AAPH). Methanol extract along with its ethyl acetate and butanol fractions showed potent free radical scavenging activity, whereas aqueous fraction was found to be devoid of any radical scavenging properties. The observed activity could be due to the higher phenolic content in the extracts (16.1, 25.29, and 17.74% w/w in methanol extract, ethyl acetate and butanol fractions respectively). HPTLC fingerprint profile of the ethyl acetate and butanol fractions were developed which would serve as reference standard for quality control of the extracts.     

Alterations in Endogenous Levels of Gibberellin-like Substances during Germination of Phaseolus vulgaris Seeds

The endogenous gibberellin-like substances were determined in mature dry and germinating bean seeds, Phaseolus vulgaris L. cv. Alabaster. Methanol extracts were partitioned against ethyl acetate and butanol at neutral and acid pH. Each phase was individually chromatographed on a silica gel column. The gibberellin activity was measured with the Tan-ginbozu dwarf rice micro-drop bioassay. Each extract was tested in two dilutions. Extracts from dry seeds showed the highest gibberellin activity, largely attributable to ethyl acetate-soluble substances. The activity was considerably reduced in extracts from seeds imbibed for 1 day. Gibberellin-like substances soluble in butanol appeared in extracts from seeds soaked for 1 or more days.     

不同品种草莓果实挥发性物质的GC-MS分析

以草莓‘晶玉’、‘甜查理’、‘晶瑶’、‘章姬’和‘丰香’等5个品种为材料,采用顶空固相微萃取-气质联用技术,分析各品种果实挥发性物质成分差异,为草莓香味育种提供依据。结果表明,5个品种果实挥发性物质中酯类均占59%以上(59.36%～77.12%);丁酸乙酯、己酸乙酯、沉香醇、呋喃酮、丁酸甲酯、己酸甲酯、2-庚酮和橙花叔醇是5品种果实中主要的特征香味物质。     

Chiral alcohol production by NADH-dependent phenylacetaldehyde reductase coupled with in situ regeneration of NADH.

Phenylacetaldehyde reductase (PAR) produced by styrene-assimilating Corynebacterium strain ST-10 was used to synthesize chiral alcohols. This enzyme with a broad substrate range reduced various prochiral aromatic ketones and beta-ketoesters to yield optically active secondary alcohols with an enantiomeric purity of more than 98% enantiomeric excess (e.e.). The Escherichia coli recombinant cells which expressed the par gene could efficiently produce important pharmaceutical intermediates; (R)-2-chloro-1-(3-chlorophenyl)ethanol (28 mg.mL-1) from m-chlorophenacyl chloride, ethyl (R)-4-chloro-3-hydroxy butanoate) (28 mg.mL-1) from ethyl 4-chloro-3-oxobutanoate and (S)-N-tert-butoxycarbonyl(Boc)-3-pyrrolidinol from N-Boc-3-pyrrolidinone (51 mg.mL-1), with more than 86% yields. The high yields were due to the fact that PAR could concomitantly reproduce NADH in the presence of 3-7% (v/v) 2-propanol in the reaction mixture. This biocatalytic process provided one of the best asymmetric reductions ever reported.     

Odoriferous compounds from the flowers of the conifers Picea abies,pinus sylvestris and Larix sibirica

A GC/MS analysis of the volatile constituents from the flowers of Norway Spruce, Picea abies, has been carried out. The volatile constituents of the female flowers were distinctly different from those of the male flowers and the twigs. Characteristic constituents are methyl and ethyl benzoate, methyl and ethyl salicylate, methyl and ethyl butanoate, borneol and bornyl acetate. In the scent from the male flowers we could only detect the same monoterpenes as in the twigs. In Larix sibirica methyl benzoate, methyl salicylate, borneol and bornyl acetate were detected in the female flowers and, in the female flowers of Pinus sylvestris, methyl salicylate was found.     

Double kinetic resolution of (±) ethyl 4 - chloro - 3- 0-acetyl butanoate

Summary Single pot double kinetic resolution of (±) ethyl 4-chloro-3-O-acetyl butanoate 1 by transesterification with n-butanol in diisopropyl ether using the enzyme Candida Cylindracea Lipase (CCL) is investigated. This technique is found to give products with better optical purity.     

Molecular docking simulation analysis of alcohol acyltransferases from two related fruit species explains their different substrate selectivities

Tropical papaya (Carica papaya) and mountain papaya (Vasconcellea pubescens) fruits are characterised for their strong and particular aroma. The aroma of both fruits is different and dominated by esters, which are synthesised by alcohol acyltransferases (AATs). The ability to produce esters is contrasting, V. pubescens (VpAAT1) being a very active enzyme towards the production of benzyl acetate, whereas C. papaya (CpAAT1) is more active towards the production of ethyl butanoate and methyl butanoate, but not benzyl acetate. In order to understand the mechanism of action at the molecular level, the structural model of CpAAT1 protein was built by comparative modelling. Conformational interaction between the protein and several ligands was carried out by molecular docking. CpAAT1 structure showed two domains connected by a large crossover loop, with a solvent channel in the centre of the structure. CpAAT1 and VpAAT1 proteins showed similar 3D structures, including their catalytic sites, but their solvent channels showed differences in size and shape. CpAAT1 solvent channel is larger, in agreement with its higher selectivity for large acyl-CoA substrates. In addition, the most favourably predicted substrate orientation in CpAAT1 was observed for methanol and butanoyl-CoA, showing a perfect coincidence with the high production rate of methyl butanoate of C. papaya fruit.     

VOLATILE FLAVOR COMPOUNDS AFFECTING CONSUMER ACCEPTABILITY OF KIWIFRUIT

Sensory research on kiwifruit has shown that specific sugars and acids, and volatile flavor compounds have causative effects on "sweet" and "acid" aroma and flavor attributes. New Zealand consumers (n = 162) assessed the perceived flavor intensity and acceptability of three volatile flavor compounds (ethyl butanoate, E-2-hexenal and hexanal) at varying levels in a model base solution. Increasing levels of ethyl butanoate positively affected all the acceptability attributes ("overall liking,""liking of aroma,""liking of flavor") as well as increased the perceived intensity of "kiwifruit aroma" and "kiwifruit flavor." Increasing levels of E-2-hexenal negatively affected all the acceptability attributes but increased the perceived intensity of "kiwifruit aroma" and "acid flavor." An increase in hexanal increased perceived intensity of "kiwifruit aroma." Ethyl butanoate and E-2-hexenel at the levels tested had the most prominent effects on consumer perceived intensity and acceptability of the "kiwifruit flavor."     

Synthesis and evaluation of a bifunctional chelate for development of Bi(III)-labeled radioimmunoconjugates

A new bifunctional ligand C-DEPA was designed and synthesized as a component for antibody-targeted radiation therapy (radioimmunotherapy, RIT) of cancer. C-DEPA was conjugated to a tumor targeting antibody, trastuzumab, and the corresponding C-DEPA-trastuzumab conjugate was evaluated for radiolabeling kinetics with ^205/6Bi. C-DEPA-trastuzumab conjugate rapidly bound ^205/6Bi, and ^205/6Bi-C-DEPA-trastuzumab conjugate was stable in human serum for 72 h. The in vitro radiolabeling kinetics and serum stability data suggest that C-DEPA is a potential chelate for preclinical RIT applications using ²¹²Bi and ²¹³Bi.     

Disruption of sexual communication in the mirid bug Lygocoris pabulinus by hexyl butanoate

1 The metathoracic scent gland in Lygocoris pabulinus contains mostly hexyl butanoate. As secretions of this gland in Heteroptera may serve as an alarm pheromone, we determined whether hexyl butanoate is released by disturbed bugs, and whether this compound disrupts sexual attraction of L. pabulinus males towards females. 2 Undisturbed males and females, and disturbed males released less than 100 ng/h hexyl butanoate, whereas disturbed females released a highly variable amount, ranging from 25 ng/h to more than 1 μg/h. 3 In the field, traps with virgin females and rubber septa containing 20 mg hexyl butanoate, caught a total of one male in a month. In control traps without hexyl butanoate, 36 males were caught in the same period. 4 In Y‐track olfactometer tests, males were not attracted to virgin females when a dispenser with 20 mg hexyl butanoate was placed in the bottle with females. Males were attracted to females when the dispenser was placed downwind from the females, but upwind from the point of male release. 5 These results suggest that males are not repelled by hexyl butanoate, but that this compound inhibits sex pheromone release in females. Application possibilities for pest management are discussed.