摘要
脂肪作为条形柄锈菌小麦专化型夏孢子萌发及侵染初期的主要营养物质,酶解产物通过乙醛酸循环转化为糖类成分供病菌生长发育所需。脂肪酶是脂肪代谢的关键酶。基于已测序的基因组序列,利用RT-PCR方法克隆了该病菌脂肪酶基因Ps LIP1序列(1302bp),编码433个氨基酸的蛋白。在毕赤酵母细胞(GS115)中成功表达Ps LIP1后,酶学活性分析显示其最适pH为8.0,最适温度为60℃,此外,发现Zn^(2+)、Cu^(2+)和Ca^(2+)对脂肪酶Ps LIP1活力有一定激活作用,Fe^(2+)、Mn^(2+)则对酶活力有抑制作用。研究结果为进一步解析脂肪酶作用机理奠定基础,同时也为从能量代谢角度制定小麦条锈病防控策略提供理论依据。
Fat is the main nutrient in the early stage of germination and infection of Puccinia striiformis f.sp.tritici(Pst)urediniospores.The enzymatic hydrolysate of fat is transformed into sugar through glyoxylate cycle for growth and development of the pathogen.Lipase is the key enzyme of fat metabolism.Based on the published genome sequence,the lipase gene Ps LIP1(1302 bp)from Pst was cloned using RT-PCR,and 433 amino acids were encoded.Ps LIP1 was successfully transformed into Pichia pastoris cells(GS115)and expressed with high activity.Enzyme activity analysis showed that the optimum pH was 8.0 and the optimum temperature was 60°C.Zn2+,Cu2+and Ca2+promoted the lipase activity of PsLIP1,while Fe2+and Mn2+inhibited the activity.The results provide a foundation for further analysis of the mechanism of lipase action and formulation of control strategies of wheat stripe rust based on perspective of energy metabolism.
作者
赵琦
李曼
康振生
汪自庆
刘杰
ZHAO Qi;LI Man;KANG Zhen-Sheng;WANG Zi-Qing;LIU Jie(State Key Laboratory of Crop Stress Biology for Arid Areas and College of Sciences,Northwest Agriculture&Forestry University,Yangling,Shaanxi 712100,China;State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences,Northwest Agriculture&Forestry University,Yangling,Shaanxi 712100,China;State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection,Northwest Agriculture&Forestry University,Yangling,Shaanxi 712100,China)
出处
《菌物学报》
CAS
CSCD
北大核心
2021年第1期152-160,共9页
Mycosystema
基金
陕西省自然科学基础研究计划(2020JZ-12)
中央高校基本科研业务费(2452019184)。
关键词
小麦条锈菌
脂肪酶
蛋白表达
理化性质
wheat stripe rust
lipase
protein expression
physical and chemical properties
