细胞色素P460的基因表达及生物学特征分析  

Gene Expression and Biological Characteristics Analysis of Cytochrome P460

作　　者：洪跃辉 范红弟 谭琼 王江海 HONG Yuehui;FAN Hongdi;TAN Qiong;WANG Jianghai(Jiangmen Key Laboratory of Traditional Chinese Medicine Ingredients and Their Mechanisms of Action/Jiangmen Engineering Technology Research Center of Health Products,Guangdong Jiangmen Chinese Medicine College,Jiangmen 529000,China;School of Marine Sciences,Sun Yat-Sen University,Zhuhai 519082,China)

机构地区：[1]广东江门中医药职业学院江门市中药成分及其作用机制重点实验室/江门市健康产品工程技术研究中心,广东江门529000 [2]中山大学海洋科学学院,广东珠海519082

出　　处：《生物化工》2024年第6期7-13,20,共8页Biological Chemical Engineering

基　　金：广东省普通高校重点领域专项(2022ZDZX4116);2022年度江门市基础与理论科学研究类科技计划项目(126);广东江门中医药职业学院2023年度院级科技计划项目和科研创新团队项目([科]JMZYYKY20232004,[科创团]JMZYYKCT20233001)。

摘　　要：目的:检测Paraburkholderia fungorum Gan-35的细胞色素P460在氨氮培养基中的基因表达,剖析该细胞色素P460的生物学特征。方法:采用荧光定量PCR检测基因表达,利用生物信息学方法分析生物学特征,分析内容包括系统进化树、理化参数、开放阅读框、亲疏水性、跨膜区、信号肽、二级结构、三级结构和功能域。结果:该细胞色素P460在氨氮培养基中的基因表达量是对照组的2.33倍,基因序列与Paraburkholderia fungorum的相应序列亲缘关系较近,含有354个氨基酸残基,属于中等分子量蛋白质,理论等电点为10.84,亲水性平均系数为0.295,含有9个开放阅读框。该P460有较多区域表现为疏水性,有4个跨膜区,缺乏信号肽,其二级结构成分包括α螺旋(49.44%)、延伸链(12.15%)、β-转角(9.04%)和无规卷曲(29.37%)。三级结构与细胞色素P450区别明显。结论:该细胞色素P460在含氨氮培养基中受到诱导表达,生物学分析结果为后续深入研究细胞色素P460的功能及其应用奠定了基础。Objective:To detect gene expression of cytochrome P460 in Paraburkholderia fungorum Gan-35 in the ammonia nitrogen culture-medium and to analyze its biological characteristics.Methods:Real-time PCR is employed to detect the gene expression.Bioinformatics methods are adopted to analyze the biological characteristics.The analysis contents include phylogenetic tree,physical and chemical parameters,open reading frame,hydrophilicity and hydrophobicity,transmembrane domain,signal peptide,secondary structure,tertiary structure and functional domain.Results:The gene expression of cytochrome P460 in the ammonia nitrogen culture-medium is 2.33 times that of the control group.Its sequence is closely related to the corresponding sequences of Paraburkholderia fungorum.It contains 354 amino acid residues and belongs to medium molecular weight proteins.Its theoretical isoelectric point and grand average of hydropathicity are 10.84 and 0.295,respectively.It contains 9 open reading frames.This P460 has more areas showing hydrophobicity,has 4 transmembrane domains,and lacks signal peptide.Its secondary structure composition includesα-helix(49.44%),extended strand(12.15%),beta turn(9.04%),and random coil(29.37%).Its tertiary structure is different from that of cytochrome P450. Conclusion: The expression of this cytochrome P460 is induced inthe ammonia nitrogen culture-medium, and the results of biological analysis lay the foundation for further studying thefunctions of cytochrome P460 and its application.

关 键 词：细胞色素P460 生物信息学 跨膜区 信号肽 三级结构 

分 类 号：Q71[生物学—分子生物学]