Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase
Sebacic acid (1,10-decanedioic acid) is an important chemical intermediate. Traditional chemical oxidation methods for sebacic acid production do not conform with “green” manufacturing. With the rapid development of enzymatic technologies, a biocatalytic cascade method based on the Baeyer–Villiger m...
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MDPI AG
2022-10-01
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| Series: | Catalysts |
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| Online Access: | https://www.mdpi.com/2073-4344/12/11/1318 |
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| author | Jie Lu Dong Lu Qiuyang Wu Shuming Jin Junfeng Liu Meng Qin Li Deng Fang Wang Kaili Nie |
| author_facet | Jie Lu Dong Lu Qiuyang Wu Shuming Jin Junfeng Liu Meng Qin Li Deng Fang Wang Kaili Nie |
| author_sort | Jie Lu |
| collection | DOAJ |
| description | Sebacic acid (1,10-decanedioic acid) is an important chemical intermediate. Traditional chemical oxidation methods for sebacic acid production do not conform with “green” manufacturing. With the rapid development of enzymatic technologies, a biocatalytic cascade method based on the Baeyer–Villiger monooxygenase was developed. The most attractive point of the method is the oleic acid that can be utilized as raw material, which is abundant in nature. However, this bio-catalysis process needs co-factor electron carriers, and the high cost of the co-factor limits its progress. In this piece of work, a co-factor in situ regeneration system between ADH from <i>Micrococcus luteus</i> WIUJH20 (<i>Ml</i>ADH) and BVMO is proposed. Since the co-factors of both enzymes are different, switching the co-factor preference of native <i>Ml</i>ADH from NAD<sup>+</sup> to NADP<sup>+</sup> is necessary. Switching research was carried out based on in silico simulation, and the sites of Tyr36, Asp 37, Ala38, and Val39 were selected for mutation investigation. The experimental results demonstrated that mutants of <i>Ml</i>ADH_D37G and <i>Ml</i>ADH_D37G/A38T/V39K would utilize NADP<sup>+</sup> efficiently, and the mutant of <i>Ml</i>ADH_D37G/A38T/V39K demonstrated the highest sebacic acid yield with the combination of BVMO. The results indicated that the in situ co-factor generation system is successfully developed, which would improve the efficiency of the biocatalytic cascade for sebacic acid production and is helpful for simplifying product isolation, thus, reducing the cost of the enzymatic transformations process. |
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| series | Catalysts |
| spelling | doaj.art-edb7c2075c0348e6b71451191c0169fe2023-11-24T04:05:24ZengMDPI AGCatalysts2073-43442022-10-011211131810.3390/catal12111318Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol DehydrogenaseJie Lu0Dong Lu1Qiuyang Wu2Shuming Jin3Junfeng Liu4Meng Qin5Li Deng6Fang Wang7Kaili Nie8College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaSebacic acid (1,10-decanedioic acid) is an important chemical intermediate. Traditional chemical oxidation methods for sebacic acid production do not conform with “green” manufacturing. With the rapid development of enzymatic technologies, a biocatalytic cascade method based on the Baeyer–Villiger monooxygenase was developed. The most attractive point of the method is the oleic acid that can be utilized as raw material, which is abundant in nature. However, this bio-catalysis process needs co-factor electron carriers, and the high cost of the co-factor limits its progress. In this piece of work, a co-factor in situ regeneration system between ADH from <i>Micrococcus luteus</i> WIUJH20 (<i>Ml</i>ADH) and BVMO is proposed. Since the co-factors of both enzymes are different, switching the co-factor preference of native <i>Ml</i>ADH from NAD<sup>+</sup> to NADP<sup>+</sup> is necessary. Switching research was carried out based on in silico simulation, and the sites of Tyr36, Asp 37, Ala38, and Val39 were selected for mutation investigation. The experimental results demonstrated that mutants of <i>Ml</i>ADH_D37G and <i>Ml</i>ADH_D37G/A38T/V39K would utilize NADP<sup>+</sup> efficiently, and the mutant of <i>Ml</i>ADH_D37G/A38T/V39K demonstrated the highest sebacic acid yield with the combination of BVMO. The results indicated that the in situ co-factor generation system is successfully developed, which would improve the efficiency of the biocatalytic cascade for sebacic acid production and is helpful for simplifying product isolation, thus, reducing the cost of the enzymatic transformations process.https://www.mdpi.com/2073-4344/12/11/1318co-factor regenerationalcohol dehydrogenasesebacic acidenzymatic cascade reactionin silico simulation |
| spellingShingle | Jie Lu Dong Lu Qiuyang Wu Shuming Jin Junfeng Liu Meng Qin Li Deng Fang Wang Kaili Nie Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase Catalysts co-factor regeneration alcohol dehydrogenase sebacic acid enzymatic cascade reaction in silico simulation |
| title | Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase |
| title_full | Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase |
| title_fullStr | Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase |
| title_full_unstemmed | Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase |
| title_short | Biocatalytic Cascade of Sebacic Acid Production with In Situ Co-Factor Regeneration Enabled by Engineering of an Alcohol Dehydrogenase |
| title_sort | biocatalytic cascade of sebacic acid production with in situ co factor regeneration enabled by engineering of an alcohol dehydrogenase |
| topic | co-factor regeneration alcohol dehydrogenase sebacic acid enzymatic cascade reaction in silico simulation |
| url | https://www.mdpi.com/2073-4344/12/11/1318 |
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