激发子蛋白SbES高密度重组表达和粉剂研发  

High-cell-density Recombinant Expression and Powder Development of the Elicitor Protein SbES

作　　者：杨扬[1] 陈奕鹏[1] 王茂存 章淑艳 刘先宝[1] 黄贵修[1] YANG Yang;CHEN Yipeng;WANG Maocun;ZHANG Shuyan;LIU Xianbao;HUANG Guixiu(Environment and Plant Protection Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Comprehensive Management of Tropical Crop Pests,Ministry of Agriculture and Rural Affairs/Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests/Hainan Engineering Research Center for Biological Control of Tropical Crops Diseases and Insect Pests,Haikou,Hainan 571101,China;Hebei Research Institute of Microbiology Company Limited,Baoding,Hebei 071051,China)

机构地区：[1]中国热带农业科学院环境与植物保护研究所/农业农村部热带作物有害生物综合治理重点实验室/海南省热带农业有害生物监测与控制重点实验室/海南省热带作物病虫害生物防治工程技术研究中心,海南海口571101 [2]河北省微生物研究所有限公司,河北保定071051

出　　处：《热带农业科学》2025年第2期64-71,共8页Chinese Journal of Tropical Agriculture

基　　金：海南省重点研发项目(No.ZDYF2021XDNY192)。

摘　　要：激发子蛋白SbES是帚枝霉属内生真菌HND5产生的一个外泌丝氨酸蛋白酶,可有效诱导多种植物产生抗病性,具有可商品化开发为植物蛋白农药的潜力。为建立该蛋白的高密度发酵及粉剂制备工艺,利用已构建好的SbES蛋白毕赤酵母异源表达菌株Pichia pastoris X-33(pRICZA::SbES),对该蛋白的高密度发酵条件、菌体破碎条件及适宜喷雾干燥条件进行优化。结果表明,在测试范围内,SbES蛋白毕赤酵母异源表达菌株在pH 6.5、温度28℃和菌体浓度180 g/L条件下诱导108 h,可获得最大表达量;目标异源表达菌株15%菌体浓度,在300 W功率下超声40 min,可获得最大的SbES蛋白量;SbES蛋白最适的助干剂为麦芽糊精,在5%麦芽糊精,干燥塔出风口温度为140℃的条件下干燥,SbES蛋白可保存最大的酶活性。本研究结果为激发子蛋白SbES的商品化开发提供了研究基础。The elicitor protein SbES is a secreted serine protease found in the endophytic fungus Sarocladium sp.HND5.This protein can effectively induce disease resistance in a variety of plants and has the potential to be commercialized as a plant protein pesticide.In this study,we used the heterologous expression strain Pichia pastoris X-33(pRICZA::SbES)to establish a high-density fermentation and powder preparation process for the protein.The results revealed that Pichia pastoris X-33(pRICZA::SbES)had the maximum SbES expression amount under the conditions at pH 6.5,28℃,and a cell concentration of 180 g/L.The heterologous expression strain could be disrupted well via 300 W ultrasonic disruption for 40 min,with a cell density of 15%.The most suitable drying agent for the SbES protein was maltodextrin.The maximum enzyme activity of the SbES protein was preserved after drying at 140℃with 5%maltodextrin.The results of this study provide a research basis for the commercial development of the elicitor protein SbES.

关 键 词：激发子蛋白 异源表达 高密度发酵 粉剂制备 植物蛋白农药 

分 类 号：S432.2[农业科学—植物病理学]