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通过异源表达 1-(4-羟苯基)-乙醇脱氢酶生产 ρ-羟基苯乙酮。

Production of ρ-Hydroxyacetophenone by Engineered Heterologously Expressing 1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase.

机构信息

Biological Engineering Laboratory, College of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, P.R. China.

Hunan Engineering Technology Research Center for Bioactive Substance Discovery of Chinese Medicine, Changsha, Hunan 410208, P.R. China.

出版信息

J Microbiol Biotechnol. 2024 Feb 28;34(2):467-475. doi: 10.4014/jmb.2310.10019. Epub 2023 Dec 8.

Abstract

ρ-Hydroxyacetophenone is an important and versatile compound that has been widely used in medicine, cosmetics, new materials, and other fields. At present, there are two ways to obtain ρ-hydroxyacetophenone. One is to extract it from plants, such as Thunb and Schneid, and the other is to synthesize it by using chemical methods. Of these two methods, the second is the main one, although it has problems, such as flammable and explosive reagents, difficult separation of by-products, and harsh reaction conditions. To solve these issues, we adopted genetic engineering in this study to construct engineered containing gene or gene. Whole-cell biotransformation was conducted under the same conditions to select the engineered with the higher activity. Orthogonal tests were conducted to determine the optimal biotransformation condition of the engineered . The results showed that the optimal condition was as follows: substrate concentration of 40 mmol/l, IPTG concentration of 0.1 mmol/l, an induction temperature of 25°C, and a transformation temperature of 35°C. Under this condition, the effects of transformation time on the ρ-hydroxyacetophenone concentration and cell growth were further studied. We found that as the transformation time extended, the ρ-hydroxyacetophenone concentration showed a gradually increasing trend. However, when the ρ-hydroxyacetophenone concentration increased to 1583.19 ± 44.34 mg/l in 24 h, cell growth was inhibited and then entered a plateau. In this research, we realized the synthesis of ρ-hydroxyacetophenone by biotransformation, and our findings lay a preliminary foundation for further improving and developing this method.

摘要

对苯二酚是一种重要且用途广泛的化合物,已广泛应用于医学、化妆品、新材料等领域。目前,获得 ρ-羟基苯乙酮有两种方法。一种是从植物中提取,如 和 ,另一种是通过化学方法合成。这两种方法中,第二种是主要方法,尽管它存在易燃和易爆试剂、副产物难以分离以及反应条件苛刻等问题。为了解决这些问题,我们在这项研究中采用了基因工程技术,构建了含有 基因或 基因的工程菌。在相同条件下进行全细胞生物转化,以选择具有更高活性的工程菌。通过正交试验确定了工程菌的最佳生物转化条件。结果表明,最佳条件如下:底物浓度为 40mmol/L,IPTG 浓度为 0.1mmol/L,诱导温度为 25°C,转化温度为 35°C。在该条件下,进一步研究了转化时间对 ρ-羟基苯乙酮浓度和细胞生长的影响。我们发现,随着转化时间的延长,ρ-羟基苯乙酮浓度呈现逐渐增加的趋势。然而,当 ρ-羟基苯乙酮浓度在 24 h 内增加到 1583.19±44.34mg/L 时,细胞生长受到抑制,然后进入平台期。在这项研究中,我们通过生物转化实现了 ρ-羟基苯乙酮的合成,为进一步改进和开发这种方法奠定了初步基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b5d/10940773/3f8e08b4f80d/jmb-34-2-467-f4.jpg

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