Production of derivatives of α-terpineol by bacterial CYP102A1 enzymes.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2024-11-25 DOI:10.1007/s10529-024-03540-w

Jeong-Hoon Kim, Chan Mi Park, Hae Chan Jeong, Sungbeom Lee, Chul-Ho Yun

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引用次数: 0

Abstract

The monooxygenase activity of engineered CYP102A1 on α-terpineol was investigated. CYP102A1 M850 mutant (F11Y/R47L/D68G/F81I/F87V/E143G/L188Q/E267V/H408R) showed the highest catalytic activity toward α-terpineol among the engineered mutants produced by random mutagenesis. The major product (P1) of α-terpineol, p-menth-1-ene-3,8-diol, was characterized by high-performance liquid chromatography, gas-chromatography mass spectrometry, and nuclear magnetic resonance spectroscopy. Three minor products (P2-P4) of α-terpineol were considered as 6-hydroxy-α,α,4-trimethyl-3-cyclohexene-1-methanol (P2), trans-sobrerol (P3), and carvone hydrate (P4). Optimal conditions for product formation were determined as pH 7.0 and 30 °C. Production of p-menth-1-ene-3,8-diol was 0.87 mM at 1 h. Structure modeling using PyMOL and CAVER Web 1.2 server indicated that several mutations of CYP102A1 M850 were involved in access tunnels and active sites, resulting in increased activity toward α-terpineol. The major product, p-menth-1-ene-3,8-diol, of α-terpineol was produced by engineered CYP102A1 M850 via regioselective carbon hydroxylation. The engineered CYP102A1 could be a suitable biocatalyst for producing α-terpineol derivatives.

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细菌 CYP102A1 酶生产 α-松油醇衍生物。

研究了工程 CYP102A1 对 α-松油醇的单加氧酶活性。在随机诱变产生的工程突变体中，CYP102A1 M850突变体（F11Y/R47L/D68G/F81I/F87V/E143G/L188Q/E267V/H408R）对α-松油醇的催化活性最高。α-松油醇的主要产物（P1）对薄荷-1-烯-3,8-二醇通过高效液相色谱法、气相色谱-质谱法和核磁共振光谱法进行了表征。α-松油醇的三种次要产物（P2-P4）被认为是 6-羟基-α,α,4-三甲基-3-环己烯-1-甲醇（P2）、反式-香叶醇（P3）和香芹酮水合物（P4）。产品形成的最佳条件被确定为 pH 值为 7.0，温度为 30 °C。使用 PyMOL 和 CAVER Web 1.2 服务器进行的结构建模表明，CYP102A1 M850 的几个突变涉及通道和活性位点，导致其对α-松油醇的活性增加。经改造的 CYP102A1 M850 通过区域选择性碳羟化作用生成了 α-松油醇的主要产物对薄荷-1-烯-3,8-二醇。经改造的 CYP102A1 可作为生产 α-松油醇衍生物的合适生物催化剂。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.