中国农业科技导报2026，Vol.28Issue(2)：145-152,8.DOI:10.13304/j.nykjdb.2025.0245
维氏气单胞菌噬菌体裂解酶在毕赤酵母中的表达及抑菌活性分析
Expression and Inhibitory Activity of Aeromonas veronii Endolysin in Pichia pastoris
摘要
Abstract
Aeromonas veronii is a common Gram-negative pathogen in aquaculture that is difficult to kill by a variety of antibiotics and antimicrobial agents due to the shielding effect of the self-produced extracellular membrane.In this study,an amino acid sequence with the N-terminal catalytic structural domain and C-terminal amphiphilic α-helical structural motif unique to the structure of Gram-negative bacterial phage endolysin was screened from the PhageScope database by bioinformatic analysis.To determine whether this amino acid sequence has antimicrobial activity,the encoding gene was optimized and synthesized according to the yeast codon and then transformed into Pichia pastoris GS115 for constitutive secretion expression,yielding recombinant A.veronii phage endolysin(LYS2)with a secretion expression level of 140 mg·L-1.The supernatant of the fermentation broth containing this endolysin exhibited significant bacteriostatic activity against A.veronii Hm091.The minimum in vitro inhibitory concentration of LYS2 against A.veronii Hm091 was 3 µg·mL-1;the optimal in vitro inhibitory conditions were 42℃and pH 4～10;the activity was stable at 4～50℃and pH 4～7;and the endolysin showed good tolerance to common metal ions and proteases.An Aeromonas veronii phage-derived endolysin with significant in vitro antimicrobial activity was experimentally verified,which provided a new antimicrobial protein with potential application for the prevention and treatment of A.veronii experimentally verified in aquaculture,and also laid a theoretical foundation for the production and application of this antimicrobial protein.关键词
维氏气单胞菌/噬菌体裂解酶/毕赤酵母/重组表达/抑菌活性Key words
Aeromonas veronii/endolysin/Pichia pastoris/recombinant expression/antimicrobial activity分类
农业科技引用本文复制引用
张晓宇,周志刚,王楠,徐时长,夏锐,药园园,冉超,张震,丁倩雯,杨雅麟..维氏气单胞菌噬菌体裂解酶在毕赤酵母中的表达及抑菌活性分析[J].中国农业科技导报,2026,28(2):145-152,8.基金项目
国家重点研发计划项目(2024YFD1300801) （2024YFD1300801）
国家自然科学基金项目(NSFC32172958,U21A20267,32330110和32172991) （NSFC32172958,U21A20267,32330110和32172991）
中国农业科学院农业科技创新工程项目(CAAS-ZDRW202305和CAAS-ASTIP-2023-IFR-05). （CAAS-ZDRW202305和CAAS-ASTIP-2023-IFR-05）
