Rational enhancement of enzyme-catalyzed enantioselective reaction by construction of recombinant enzymes based on additive strategy

Han, Yu; Zhou, Xiaoxue; Zheng, Liangyu

doi:10.1007/s00449-019-02170-1

Cited by 4 publications

(3 citation statements)

References 35 publications

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“…SDS‐PAGE analysis of the broken bacterial lysate confirmed that some constructed n His‐CalB‐10Lys such as 2His‐CalB‐10Lys, 4His‐CalB‐10Lys, 6His‐CalB‐10Lys and 8His‐CalB‐10Lys could be expressed as soluble proteins. For 6His‐CalB‐ n Lys such as 6His‐CalB and 6His‐CalB‐5Lys, they were expressed mainly as an inclusion body form 34 . Although the soluble r CalBs could be easily obtained, they could not be purified using a nickel affinity column, which was unfavorable for the deep study of the r CalB‐catalyzed MBH reactions.…”

Section: Resultsmentioning

confidence: 99%

“…For 6His-CalB-nLys such as 6His-CalB and 6His-CalB-5Lys, they were expressed mainly as an inclusion body form. 34 Although the soluble rCalBs could be easily obtained, they could not be purified using a nickel affinity column, which was unfavorable for the deep study of the rCalB-catalyzed MBH reactions. In view of the commercial immobilized Novozym 435 applied in our previous lipase/amide co-catalyzed system, 32 the immobilized rCalBs were prepared in our laboratory by bioinspired immobilization technology.…”

Section: Resultsmentioning

confidence: 99%

“…This study is started from our previously established lipase CalB/amide co‐catalyzed MBH reaction between 4‐nitrobenzaldehyde and 2‐cyclohexen‐1‐one 32 . Combined with our previous findings that the lipase CalB‐catalyzed enantioselective hydrolysis can be enhanced by construction of the recombinant CalBs ( r CalBs) with polyamino acid tails, 33,34 it allows us to consider, since both the amino acids fused on CalB and the amide additive in CalB/amide co‐catalyzed MBH reaction belong to amine compounds, that the lipase r CalBs may produce positive effects on the MBH reactions. According to this assumption, the r CalBs are first attempted to be used in the MBH reaction to observe their catalytic performance.…”

Section: Introductionmentioning

confidence: 99%

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Engineering the simply asymmetric Morita–Baylis–Hillman reactions in a sodium phosphate buffer system: from enzyme‐ to proline‐catalyzed transformation

Cui

Gao

et al. 2022

J of Chemical Tech & Biotech

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BACKGROUND The Morita–Baylis–Hillman (MBH) reaction is an important carbon–carbon bond‐forming reaction. However, most MBH reactions suffer disadvantages such as high amounts of catalyst and hazardous solvents, long reaction time, low reactivity and selectivity. In this study, simple, cost‐effective and eco‐friendly asymmetric MBH reactions are developed. RESULTS Following our previous study that the commercial Candida antarctica lipase B (lipase CalB)‐catalyzed MBH reaction can be activated by introducing an amide compound as a co‐catalyst, the recombinant lipase CalBs (rCalBs) with polyamino acid tails are first attempted to be used in the same MBH reaction, but unfortunately chiral MBH products are not obtained. Enoate reductase (ER) is found to give asymmetric transformation of the MBH reaction with a conversion of 36.9% and enantiomeric excess (e.e.p) of 6.8% when other enzyme resources are expanded. According to the observation that the surface amino acids of ER may be key factors in achieving the MBH reaction, single amino acids are subsequently selected to catalyze the MBH reactions, and the reverse configuration of (R)‐ and (S)‐MBH products with e.e.p of 63.9% and 62.1% for single l‐proline‐ and d‐proline‐catalyzed MBH reactions are reached only in 4 h, respectively. CONCLUSION Proline can afford the asymmetric MBH reaction in a sodium phosphate buffer only in 4 h without the help of the other co‐catalyst in our study, which is superior to those reported. The study aims to introduce a novel phenomenon observed from enzyme‐catalyzed to proline‐catalyzed MBH reactions. © 2022 Society of Chemical Industry (SCI).

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%