桂花CCD4的过表达及体外β-胡萝卜素合成β-离子酮的研究

Q2 Chemical Engineering

Journal of Molecular Catalysis B-enzymatic Pub Date : 2016-12-01 DOI:10.1016/j.molcatb.2016.10.003

Xuesong Zhang , Jianjun Pei , Linguo Zhao , Feng Tang , Xianying Fang , Jingcong Xie

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引用次数: 21

摘要

本研究以桂花为原料，利用大肠杆菌在不同的细菌生长条件下制备了重组类胡萝卜素裂解双加氧酶4，并建立了以β-胡萝卜素为原料制备天然β-离子酮的生物技术方法。用高效液相色谱法分析β-离子酮，并根据β-离子酮的浓度变化测定OfCCD4活性。在pH 8.0和35℃条件下，纯化后的重组蛋白在1 h内活性最高，为14.3 U/mg， β-离子酮浓度最高，为71.186 mg/L，添加9% Triton X-100和2%液体石蜡后，酶活性和β-离子酮浓度均可提高近6倍。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Overexpression and characterization of CCD4 from Osmanthus fragrans and β-ionone biosynthesis from β-carotene in vitro

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Overexpression and characterization of CCD4 from Osmanthus fragrans and β-ionone biosynthesis from β-carotene in vitro

In this study, a recombinant carotenoid cleavage dioxygenase 4 was produced from Osmanthus fragrans by E. coli under different bacterial growth conditions and used to develop a biotechnological method for preparation of natural β-ionone from β-carotene. β-ionone was analyzed by HPLC and OfCCD4 activity was measured based on concentration change of β-ionone. At pH 8.0 and 35 °C, the greatest activity of the purified recombinant protein was 14.3 U/mg and the maximum concentration of β-ionone was 71.186 mg/L within 1 h. Both the enzyme activity and the concentration of β-ionone could increase by nearly 6 times with addition of 9% Triton X-100 and 2% liquid paraffin.

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来源期刊

Journal of Molecular Catalysis B-enzymatic 生物-生化与分子生物学

CiteScore

2.58

自引率

0.00%

发文量

审稿时长

3.4 months

期刊介绍： Journal of Molecular Catalysis B: Enzymatic is an international forum for researchers and product developers in the applications of whole-cell and cell-free enzymes as catalysts in organic synthesis. Emphasis is on mechanistic and synthetic aspects of the biocatalytic transformation. Papers should report novel and significant advances in one or more of the following topics; Applied and fundamental studies of enzymes used for biocatalysis; Industrial applications of enzymatic processes, e.g. in fine chemical synthesis; Chemo-, regio- and enantioselective transformations; Screening for biocatalysts; Integration of biocatalytic and chemical steps in organic syntheses; Novel biocatalysts, e.g. enzymes from extremophiles and catalytic antibodies; Enzyme immobilization and stabilization, particularly in non-conventional media; Bioprocess engineering aspects, e.g. membrane bioreactors; Improvement of catalytic performance of enzymes, e.g. by protein engineering or chemical modification; Structural studies, including computer simulation, relating to substrate specificity and reaction selectivity; Biomimetic studies related to enzymatic transformations.