Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate
Monoethyl adipate (MEA) is a highly valuable monoester for activating resistance mechanisms and improving protective effects in pathogen-attacked plants. The cutinase ACut2 from the non-conventional yeast <i>Blastobotrys (Arxula) raffinosifermentans (adeninivorans)</i> was used for its s...
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| Format: | Article |
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MDPI AG
2022-06-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/7/1316 |
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| author | Marion Rauter Daniela Nietz Gotthard Kunze |
| author_facet | Marion Rauter Daniela Nietz Gotthard Kunze |
| author_sort | Marion Rauter |
| collection | DOAJ |
| description | Monoethyl adipate (MEA) is a highly valuable monoester for activating resistance mechanisms and improving protective effects in pathogen-attacked plants. The cutinase ACut2 from the non-conventional yeast <i>Blastobotrys (Arxula) raffinosifermentans (adeninivorans)</i> was used for its synthesis by the desymmetrization of dicarboxylic acid diester diethyl adipate (DEA). Up to 78% MEA with 19% diacid adipic acid (AA) as by-product could be synthesized by the unpurified ACut2 culture supernatant from the <i>B. raffinosifermentans</i> overexpression strain. By adjusting pH and enzyme concentration, the selectivity of the free ACut2 culture supernatant was increased, yielding 95% MEA with 5% AA. Selectivity of the carrier immobilized ACut2 culture supernatant was also improved by pH adjustment during immobilization, as well as carrier enzyme loading, ultimately yielding 93% MEA with an even lower AA concentration of 3–4%. Thus, optimizations enabled the selective hydrolysis of DEA into MEA with only a minor AA impurity. In the up-scaling, a maximum of 98% chemical and 87.8% isolated MEA yield were obtained by the adsorbed enzyme preparation with a space time yield of 2.6 g L<sup>−1</sup> h<sup>−1</sup>. The high monoester yields establish the ACut2-catalyzed biosynthesis as an alternative to existing methods. |
| first_indexed | 2024-03-09T10:14:28Z |
| format | Article |
| id | doaj.art-7cac91fdbf7c488f800bdf35711e8272 |
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| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T10:14:28Z |
| publishDate | 2022-06-01 |
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| series | Microorganisms |
| spelling | doaj.art-7cac91fdbf7c488f800bdf35711e82722023-12-01T22:28:12ZengMDPI AGMicroorganisms2076-26072022-06-01107131610.3390/microorganisms10071316Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl AdipateMarion Rauter0Daniela Nietz1Gotthard Kunze2Orgentis Chemicals GmbH, Bahnhofstr. 3–5, Gatersleben, D-06466 Stadt Seeland, GermanyLeibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, Gatersleben, D-06466 Stadt Seeland, GermanyLeibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, Gatersleben, D-06466 Stadt Seeland, GermanyMonoethyl adipate (MEA) is a highly valuable monoester for activating resistance mechanisms and improving protective effects in pathogen-attacked plants. The cutinase ACut2 from the non-conventional yeast <i>Blastobotrys (Arxula) raffinosifermentans (adeninivorans)</i> was used for its synthesis by the desymmetrization of dicarboxylic acid diester diethyl adipate (DEA). Up to 78% MEA with 19% diacid adipic acid (AA) as by-product could be synthesized by the unpurified ACut2 culture supernatant from the <i>B. raffinosifermentans</i> overexpression strain. By adjusting pH and enzyme concentration, the selectivity of the free ACut2 culture supernatant was increased, yielding 95% MEA with 5% AA. Selectivity of the carrier immobilized ACut2 culture supernatant was also improved by pH adjustment during immobilization, as well as carrier enzyme loading, ultimately yielding 93% MEA with an even lower AA concentration of 3–4%. Thus, optimizations enabled the selective hydrolysis of DEA into MEA with only a minor AA impurity. In the up-scaling, a maximum of 98% chemical and 87.8% isolated MEA yield were obtained by the adsorbed enzyme preparation with a space time yield of 2.6 g L<sup>−1</sup> h<sup>−1</sup>. The high monoester yields establish the ACut2-catalyzed biosynthesis as an alternative to existing methods.https://www.mdpi.com/2076-2607/10/7/1316diethyl adipatemonoethyl adipatecutinase 2 (ACut2)desymmetrization<i>Blastobotrys raffinosifermentans</i> |
| spellingShingle | Marion Rauter Daniela Nietz Gotthard Kunze Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate Microorganisms diethyl adipate monoethyl adipate cutinase 2 (ACut2) desymmetrization <i>Blastobotrys raffinosifermentans</i> |
| title | Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate |
| title_full | Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate |
| title_fullStr | Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate |
| title_full_unstemmed | Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate |
| title_short | Cutinase ACut2 from <i>Blastobotrys</i><i>raffinosifermentans</i> for the Selective Desymmetrization of the Symmetric Diester Diethyl Adipate to the Monoester Monoethyl Adipate |
| title_sort | cutinase acut2 from i blastobotrys i i raffinosifermentans i for the selective desymmetrization of the symmetric diester diethyl adipate to the monoester monoethyl adipate |
| topic | diethyl adipate monoethyl adipate cutinase 2 (ACut2) desymmetrization <i>Blastobotrys raffinosifermentans</i> |
| url | https://www.mdpi.com/2076-2607/10/7/1316 |
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