中国水产科学2025，Vol.32Issue(6)：729-741,13.DOI:10.12264/JFSC2025-0085
南极磷虾胰蛋白酶基因克隆、异源表达及其降解塑化剂研究
Cloning and heterologous expression of Antarctic krill trypsin gene and its degradation of plasticizer
摘要
Abstract
The objective of this study was to obtain the tryptic albuminase gene EsTRY from Antarctic krill(Euphausia superba)by screening and cloning,construct the recombinant expression vector EsTRY-pCold-SUMO,realize the high efficiency of heterologous expression in Escherichia coli,and reveal the degradation activity of this enzyme on phthalate esters(PAEs).The gene of trypsin EsTRY of Antarctic krill is 1350 bp in length,encoding 449 amino acids,with a theoretical molecular weight(MW)of 47040.35 Da and a theoretical isoelectric point(pI)of 4.90.Structural analysis revealed the presence of a catalytic triad(Asp/Glu-Ser-His)and a substrate-specific binding site,classifying EsTRY as a member of the serine protease family.The esterase activity of EsTRY was determined to be 18973 U/mL,with a specific enzyme activity of 5802.15 U/mg.The optimal reaction temperature was found to be 25 ℃,and the optimal pH was determined to be 7.0.Enzyme-catalyzed degradation experiments demonstrated that EsTRYpossessed the capacity to degrade phthalic acid esters(PAEs),with 13.2％,34.4％,and 63.5％of the 1 mmol/L of PAEs being degraded within 15 min,respectively.Furthermore,48.9％of 1 mmol/L DMP,DEP,DPRP,and DBP were degraded within 15 min,and the degradation rates reached 39.4％,62.7％,86.8％,and 98.7％within 5 h,respectively.In this paper,we investigated the degradation activity of EsTRY on PAEs,which provides a new potential tool for the biological management of plasticizer pollution,as well as a scientific basis for the efficient utilization and high-value exploitation of Antarctic krill.关键词
南极磷虾/胰蛋白酶/异源表达/PAEs/降解/塑化剂Key words
Antarctic krill/trypsin/heterologous expression/degradation of plasticisers分类
农业科技引用本文复制引用
邹华英,谭佳豪,王芳,宋佳,陈梦瑶,徐甲坤..南极磷虾胰蛋白酶基因克隆、异源表达及其降解塑化剂研究[J].中国水产科学,2025,32(6):729-741,13.基金项目
国家重点研发计划项目(2022YFC2807500). （2022YFC2807500）
