The industrial application of biocatalysis for the production of natural flavour compounds is illustrated by a discussion of the production of vanillin, gamma-decalactone, carboxylic acids, C6 aldehydes and alcohols ('green notes'), esters, and 2-phenylethanol. Modern techniques of molecular biology and process engineering, such as heterologous expression of genes, site-directed mutagenesis, whole-cell biocatalysis in biphasic systems, and cofactor regeneration for in vitro oxygenation, may result in more biocatalytic processes for the production of flavour compounds in the future.
2-Phenylethanol (2-PE) is an important flavour and fragrance compound with a rose-like odour. Most of the world's annual production of several thousand tons is synthesised by chemical means but, due to increasing demand for natural flavours, alternative production methods are being sought. Harnessing the Ehrlich pathway of yeasts by bioconversion of L-phenylalanine to 2-PE could be an option, but in situ product removal is necessary due to product inhibition. This review describes the microbial production of 2-PE, and also summarizes the chemical syntheses and the market situation.
An integrated bioprocess for the production of the natural rose-like aroma compounds, 2-phenylethanol (2-PE) and 2-phenylethylacetate (2-PEAc), from L-phenylalanine (L-phe) with yeasts was investigated. The hydrophobicity of the products leads to product inhibition, which can be compensated by in situ product removal (ISPR). An organophilic pervaporation unit, equipped with a polyoctylmethylsiloxane (POMS) membrane, was coupled via a bypass to a bioreactor and proved to be a suitable technique for the in situ removal of high-boiling products from culture broth. With batch cultures of the thermotolerant yeast Kluyveromyces marxianus CBS 600 in a standard medium at 35 degrees C, the use of pervaporation resulted in a double 2-PE concentration (2.2 g/L) and 1.3 g/L 2-PEAc, which only accumulated transiently in low concentrations during cultivation without ISPR. Using a previously optimized medium, the variation of the temperature from 30 degrees C to 40 degrees C caused an increase in the total conversion yield from 63% to 79%, corresponding to total product concentrations of 5.23 and 5.85 g/L, respectively. In the 40 degrees C batch experiment, the volumetric productivity (2-PE + 2-PEAc) during the exponential phase was 5.2 mmol/L h. While for 2-PE, there is still potential for further optimization, the more hydrophobic 2-PEAc was nearly completely removed from the aqueous culture broth (enrichment factor >400), resulting in highly aroma-enriched permeates. Due to the temperature-correlated performance of the pervaporation, the bioconversion was still efficient even at 45 degrees C (conversion yield: 69%). Surprisingly, at 45 degrees C, the molar ratio of the two products inverted and 2-PEAc turned out to be the main product (4.0 g/L), which opens easy control of the reaction's selectivity by external means. Retrofitting the process with interim heating and cooling equipment to use different temperature levels for cultivation and pervaporation resulted in a decreased yield and product concentration caused by multiple stress factors. The medium composition affected the pervaporation efficiency with molasses acting detrimental.
Fourteen yeast strains were screened for production of 2-phenylethanol from L-phenylalanine with molasses as carbon source. Up to 1 g 2-phenylethanol l-1 was obtained. Using oleyl alcohol as a second phase for in situ product removal to enhance the production of 2-phenylethanol increased the yield to about 3 g 2-phenylethanol l-1 at 35 degrees C. The most productive strains were Kluyveromyces marxianus CBS 600 and CBS 397.
Good governance practices through electronic government (eGov) platforms can be suitable instruments for strengthening the outcomes of smart city policies. While eGov is the application of information and communication technologies to public services, good governance defines how well public authorities manage public and social resources. Contemporary public management views, such as ‘new public service’, include citizen participation as a critical factor to sustainable government in smart cities. Public services, in the age of digital technology, need to not only be delivered through eGov platforms, but also need to be coproduced with the engagement of social players, e.g., citizens. In this sense, eGov platforms act as digital commons, and conceived as digital spaces, where citizens and public agents interact and collaborate. In this paper, we presented the Municipal eGov Platform Assessment Model (MEPA), which is a model specifically developed to evaluate eGov platforms regarding their potential to promote commons in smart cities. The study applied MEPA to 903 municipal websites across Brazil. The results revealed that the majority of investigated Brazilian eGov platforms have only a low level of digital commons maturity. This finding discloses less citizenship coproduction, and fewer opportunities for city smartness. As the MEPA model offers public authorities an instrument to depict weaknesses and strengths of municipal eGov platforms, its adoption provides an opportunity for authorities to plan and manage their platforms to act as promoters of digital commons and citizen coproduction.
Selective transformations of limonene by asco- and basidiomycetes were investigated. On the shake flask scale, Penicillium citrinum hydrated R-(+)-limonene to α-terpineol [83% regioselectivity (rs), more than 80 mg 1-1 product yield], and Gongronella butleri catalysed the terminal oxidation to yield perillyl alcohol (60% rs, 16 mg 1-1). On the laboratory bioreactor scale, Penicillium digitatum produced a peak concentration of 506 mg α-terpineol 1-1 in the fed-batch mode, equivalent to a theoretical yield of 67%, and no volatile by-products were found. Fusarium proliferatum transformed R-(+)-limonene enantiospecifically to cis-(+)- carveol (98.6% ee, more than 35 mg 1-1 product yield) and S-(-)-limonene predominantly to trans-(-)-carveol (96.3% ee). Pleurotus sapidus selectively dehydrogenised the accumulating trans-(-)-carveol to the corresponding enantiopure R-(-)-carvone. The results show that a careful selection of strain and bioprocess parameters may improve both the yield and the optical purity of a desired product.
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