Kinetic analysis and modelling of enzymatic (R)-phenylacetylcarbinol batch biotransformation process

Leksawasdi, Noppol; Chow, Yvonne; Breuer, Michael; Hauer, Bernhard; Rosche, Bettina; Rogers, Peter L.

doi:10.1016/j.jbiotec.2004.04.001

Cited by 23 publications

(64 citation statements)

References 25 publications

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“…Metabolically, the efficiency of ethanol production process depends not only on PDC activity but also alcohol dehydrogenase activity as it is a two steps bioconversion process of pyruvate to acetaldehyde and acetaldehyde to ethanol (Candy & Duggleby, ). The PAC production process, on the other hand, is a carboligation reaction of joining active acetaldehyde being produced from pyruvate to benzaldehyde at the active site of PDC in which PAC is released as a product (Leksawasdi et al, ). Based on above reasons, one might thus propose that ethanol production level can be used as an indication for the presence of PDC activity to a certain degree but the ethanol level being produced might not necessarily correspond to PAC production level.…”

Section: Resultsmentioning

confidence: 99%

“…The benzoic acid concentration levels were detected during biotransformation in range of 1.56 ± 0.62–3.94 ± 0.31 mM with whole cells of C. tropicalis TISTR 5306 while higher levels of this acid (2.04 ± 0.86–6.83 ± 0.84 mM) were detected for S. cerevisiae TISTR 5606. Benzoic acid was generated through oxidation process of benzaldehyde with dissolved oxygen in the buffering system (Leksawasdi et al, ). The other by‐products (acetaldehyde, acetoin, and benzyl alcohol), were not detected in this reaction system.…”

Section: Resultsmentioning

confidence: 99%

“…Biotransformation experiment was carried out for whole cells of both yeast strains collected at 48 hr cultivation period in an equal volume of 15:15 ml bi‐phasic system (adapted from Gunawan, Breuer, Hauer, Rogers, & Rosche, ) based on similar system described by Leksawasdi et al () and Agustina et al (). The aqueous phase contained 240 mM sodium pyruvate in 1 M phosphate buffer (pH 6.5/10 M KOH) (Khemacheewakul et al, ) with 1 mM MgSO 4 .7H 2 O and 1 mM thiamine pyrophosphate (TPP) as cofactors (Leksawasdi et al, ; Leksawasdi, Rogers, & Rosche, ). Palm oil (1.03 USD/l, AT‐Science, ()) was used as an organic phase with the presence of 200 mM benzaldehyde.…”

Section: Methodsmentioning

confidence: 99%

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Ethanol and phenylacetylcarbinol production processes of Candida tropicalis TISTR 5306 and Saccharomyces cerevisiae TISTR 5606 in fresh juices from longan fruit of various sizes

Nunta

Techapun

Kuntiya

et al. 2018

J Food Process Preserv

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Fresh juice from C grade longan (mg/g fresh fruit; total sugars 51.6 ± 0.5, nitrogen 0.021 ± 0.004; gallic acid 0.025 ± 0.001; ellagic acid 0.016 ± 0.001) with addition of 8.52 g ammonium sulfate/l was the optimal medium for cultivation processes of Saccharomyces cerevisiae TISTR 5606 and Candida tropicalis TISTR 5306 capable of producing ethanol and phenylacetylcarbinol (PAC) among five longan grades. S. cerevisiae TISTR 5606 produced ethanol (33.4 ± 3.2 g/L) at a significantly higher level (p ≤ 0.05) than C. tropicalis TISTR 5306 (22.3 ± 1.1 g/L) with similar ethanol yields (Yp/s) between 0.21 and 0.22 g ethanol produced/g sugars consumed. Whole cells of C. tropicalis TISTR 5306 produced a significantly higher (p ≤ 0.05) PAC level (27.2 ± 0.7 mM) than S. cerevisiae TISTR 5606 (3.59 ± 1.33 mM) after 6 hr in an equivalent volume biphasic biotransformation system. Practical applications Longan is one of the important economic fruit of Thailand with production volume in 2017 reaching one million tons. Less than 10% of these are domestically consumed while more than 90% are exported to several countries whose population frequently consume longan as a nutritious food supplement. This study can help solving overproduction problem of longan fruit by processing C grade fresh longan, which is accounted for 5% of the overall production volume, in the form of juice extract with the relatively high sugars content. Microbial fermentation of this juice and subsequent whole cells biotransformation process result in ethanol and PAC, respectively. Ethanol can be used as an alternative biofuel or important industrial solvent while PAC is a precursor for production of commercial nasal decongestant (ephedrine) or anti‐asthmatic compound (pseudoephedrine).

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Ethanol and phenylacetylcarbinol production processes of Candida tropicalis TISTR 5306 and Saccharomyces cerevisiae TISTR 5606 in fresh juices from longan fruit of various sizes

Nunta

Techapun

Kuntiya

et al. 2018

J Food Process Preserv

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“…Pyruvate decarboxylase (PDC) is the crucial penultimate tetrameric enzyme in the ethanol formation process with the role of cleaving a carbon dioxide molecule from pyruvate through a decarboxylation catalytic step resulting in an active acetaldehyde (Leksawasdi et al, ; Leksawasdi, Rogers, & Rosche, ; Rosche et al, ). The proton consuming step of carbo‐ligation reaction between active acetaldehyde and added PDC deactivating substrate—benzaldehyde (Khemacheewakul et al, ; Leksawasdi, Breuer, Hauer, Rosche, & Rogers, )—on the active sites of PDC would result in the formation of a precursor— R ‐phenylacetylcarbinol (PAC, C 9 H 10 O 2 , MW 150 g/mol)—for production of well‐known diastereomeric commercial bronchial dilators such as ephedrine and pseudoephedrine (both C 10 H 15 NO, MW 165 g/mol) (Agustina et al, ; Leksawasdi, Rosche, & Rogers, ; PubChem, , , ).…”

Section: Introductionmentioning

confidence: 99%

Batch and continuous cultivation processes of Candida tropicalis TISTR 5306 for ethanol and pyruvate decarboxylase production in fresh longan juice with optimal carbon to nitrogen molar ratio

Nunta

Techapun

Jantanasakulwong

et al. 2019

J Food Process Engineering

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The carbon to nitrogen (C/N) molar ratio of 21.88 ± 0.20 in fresh juice from C‐grade longan (FLJ‐C) supplemented with ammonium sulfate (AMS) was optimal for cultivation of Candida tropicalis TISTR 5306 producing ethanol concentration of 24.0 ± 1.1 g/L and specific pyruvate decarboxylase (PDC) activity of 0.138 ± 0.001 U/mg protein. The highest ethanol concentration in a 10 L continuous process (34.3 ± 0.5 g/L, dilution rate [D] = 0.0492 hr−1) was significantly higher (p ≤ .05) than that in 100 L batch process (13.2 ± 0.2 g/L at 108 hr). The highest volumetric PDC activities of 0.081 ± 0.001 U/ml (D = 0.0070 hr−1) and 0.107 ± 0.001 U/ml (48 hr) were attained for continuous and batch processes, respectively. The affinity constants and growth‐related weighting factors on sucrose and monosaccharides (glucose and fructose with the same values) utilization were (123 ± 1 g/L, 0.14 ± 0.01) and (34.1 ± 1.5 g/L, 0.43 ± 0.01) with MS and R2 of 92.2 and .9552, respectively. Practical applications FLJ‐C is an example of agricultural produces with relatively high production quantity (50,000 t/year in Thailand) and high sugars content but low marketing demands due to the smallest size of pulp. The complex nitrogen source contains costly components (yeast extract—78.3 USD/kg, malt extract—148 USD/kg, and peptone—151 USD/kg) which can be replaced with an inexpensive AMS (1.05 USD/kg). Further consideration of C/N molar ratio in FLJ‐C can optimize growth, ethanol (alternative biofuel/industrial solvent) production, and specific PDC (biocatalyst for phenylacetylcarbinol—PAC, anti‐asthmatic and nasal decongestant precursors) activity of C. tropicalis TISTR 5306 whole cells which can be used as a case study for other related yeasts. Batch and continuous production at 100 L and 10 L also illustrate practical utilization of FLJ‐C with AMS supplement at larger scales for development of subsequent zero waste process.

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“…Fermenting yeasts such as Saccharomyces cerevisiae have isoenzymes of pyruvate decarboxylase (PDC) which are capable of carboligation. Typically, carboligation involves the condensation of two molecules of acetaldehyde to produce acetoin [1]. In this biotransformation, acetaldehyde (formed from the decarboxylation of pyruvate which is a product of glycolysis) condenses with exogenously added benzaldehyde to form (R)-PAC.…”

Section: Introductionmentioning

confidence: 99%

(R)-PAC Biosynthesis in [BMIM][PF6]/Aqueous Biphasic System Using Saccharomyces cerevisiae BY4741 Cells

Kandar

Suresh

Noronha

2014

Appl Biochem Biotechnol

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(R)-phenylacetylcarbinol or (R)-PAC is a pharmaceutical precursor of (1R, 2S) ephedrine and (1S, 2S) pseudoephedrine. Biotransformation of benzaldehyde and glucose by pyruvate decarboxylase produces (R)-PAC. This biotransformation suffers from toxicity of the substrate, product [(R)-PAC] and by-product (benzyl alcohol). In the present study, ionic liquid/aqueous biphasic system was employed to enhance (R)-PAC production. Fermented broth was the reaction medium in which Saccharomyces cerevisiae BY4741 was the source of pyruvate decarboxylase. Hydrophobic ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]) was the non-aqueous phase in which toxic compounds reside. Biocompatibility of [BMIM][PF6] and adequate distribution coefficients of benzaldehyde, (R)-PAC and benzyl alcohol were determined. A Box-Behnken design and response surface methodology were used for the optimization of biotransformation variables in order to maximize (R)-PAC yield and productivity. The results showed higher (R)-PAC yield and productivity of ∼1.5-fold each in the biphasic biotransformation of phase volume ratio 0.05 as compared to the monophasic (conventional) biotransformation. Moreover, the level of major by-product benzyl alcohol was also 3.5-fold lower in biphasic biotransformation. [BMIM][PF6]/aqueous biphasic system is a new approach which could intensify the (R)-PAC production.

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Kinetic analysis and modelling of enzymatic (R)-phenylacetylcarbinol batch biotransformation process

Cited by 23 publications

References 25 publications

Ethanol and phenylacetylcarbinol production processes of Candida tropicalis TISTR 5306 and Saccharomyces cerevisiae TISTR 5606 in fresh juices from longan fruit of various sizes

Ethanol and phenylacetylcarbinol production processes of Candida tropicalis TISTR 5306 and Saccharomyces cerevisiae TISTR 5606 in fresh juices from longan fruit of various sizes

Batch and continuous cultivation processes of Candida tropicalis TISTR 5306 for ethanol and pyruvate decarboxylase production in fresh longan juice with optimal carbon to nitrogen molar ratio

(R)-PAC Biosynthesis in [BMIM][PF6]/Aqueous Biphasic System Using Saccharomyces cerevisiae BY4741 Cells

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