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Use of bacteria, proteins or peptides derived from clones of Bacillus subtilis, deposited at american type culture collection, as stimulators of entire natural plant, against pathogen of fungal, bacterial and/or viral type cultures
Use of bacteria, proteins or peptides derived from clones of Bacillus subtilis, deposited at american type culture collection, as stimulators of entire natural plant, against pathogen of fungal, bacterial and/or viral type cultures
Use of bacteria, proteins or peptides derived from: clones of Bacillus subtilis, deposited at ATCC (american type culture collection) under number of 55614, and 6633; clones of Bacillus cereus, deposited at ATCC under number of 21928, 55609, 21929, 53522 and 55609; and clones of Bacillus pumilus, deposited at ATCC under numbers of 31132 and 31340, and at northern regional research laboratory collection under number of B-30087, as stimulators of entire natural plant, against pathogen of fungal, bacterial and/or viral type cultures, is claimed. An independent claim is included for a process for obtaining a composition containing bacteria, proteins or peptides, comprising: culturing the bacteria using a culture medium, which is relatively simple (MS), where the composition comprises potassium dihydrogen phosphate (2.5 g/l), dipotassium dihydrogen phosphate (2.5 g/l), diammonium hydrogen phosphate ((NH 4) 2HPO 4) (1 g/l), magnesium sulfate heptahydrate (0.2 g/l), manganese sulfate heptahydrate (7 mg/l), ferrous sulfate heptahydrate (0.01 g/l), and calcium chloride (10 mg/l), adding standard saline solution of 1% (p/v) D-glucose and adjusting the pH 7, sterilizing the medium by autoclaving at 120[deg] C for 20 minutes, inoculating a preculture of bacteria in MS medium for 15 hours at 30[deg] C under agitation, and placing a flask containing the bacteria in culture at 37[deg] C under gentle rotative agitation (150 rpm) for 5-7 days; and (b) separating bacteria and proteins, performed at 4[deg] C comprising centrifuging the contents of each flask at 4500 rpm for 30 minutes, washing the pellet in 1M sodium chloride solution, twice with distilled water containing 1 mM phenylmethylsulfonyl fluoride and 10 mM EDTA and then resuspended in a volume of demineralized water and purifying the protein part of a discontinuous sucrose gradient by method of Thomas and Ellar (1983) [ultracentrifugation at 25000 rpm and 48[deg] C for 16 hours], concentrating into protein, which is determined by the Bradford method after alkaline solubilization of the extract and electrophoresis (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) denaturing conditions for detecting proteins using Thomas and Ellar (1983) method. ACTIVITY : Fungicide; Virucide; Antibacterial. MECHANISM OF ACTION : None given.
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机译：来源于以下细菌，蛋白质或肽的用途：枯草芽孢杆菌的克隆，保藏在ATCC（美国典型培养物保藏中心），编号为55614和6633;蜡样芽胞杆菌的克隆，保藏在ATCC，编号为21928、55609、21929、53522和55609;和短小芽孢杆菌的克隆，它们作为抗氧化剂，细菌和/或病毒型培养物的病原体，作为完整的天然植物的刺激物，保藏在ATCC，编号为31132和31340，并存放在北部区域研究实验室，编号为B-30087，被要求。包括获得包含细菌，蛋白质或肽的组合物的方法的独立权利要求，所述方法包括：使用相对简单的培养基（MS）培养细菌，其中所述组合物包含磷酸二氢钾（2.5 g / l） ，磷酸二氢二钾（2.5 g / l），磷酸氢二铵（（NH 4）2HPO 4）（1 g / l），七水硫酸镁（0.2 g / l），七水硫酸锰（7 mg / l），亚铁硫酸七水合物（0.01 g / l）和氯化钙（10 mg / l），添加1％（p / v）D-葡萄糖的标准盐溶液并调节pH 7，通过在120°高压灭菌对培养基进行灭菌在70℃下搅拌20分钟，在搅拌下于30℃下在MS培养基中将细菌预培养15小时，然后将含有细菌的烧瓶在37℃下在缓慢旋转搅拌（150rpm）下培养5分钟。 -7天; （b）分离细菌和蛋白质，在4℃下进行，包括将每个烧瓶中的内容物在4500 rpm下离心30分钟，在1M氯化钠溶液中洗涤沉淀，用含有1 mM苯基甲基磺酰氟和10的蒸馏水洗涤两次。 mM EDTA，然后重悬于一定量的软化水中，并通过Thomas和Ellar（1983）[在25000 rpm和48°C下超速离心16小时]的方法纯化不连续蔗糖梯度的蛋白质部分，浓缩为蛋白质，提取液经碱溶和电泳（十二烷基硫酸钠-聚丙烯酰胺凝胶电泳）变性条件后用Bradford方法测定，使用Thomas和Ellar（1983）方法检测蛋白质的变性条件。活动：杀菌剂;杀病毒剂;抗菌。作用机理：未给出。
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