以透明圈法、Azocoll法和16S r DNA,分离及鉴定耐高温、嗜中性和产胶原蛋白酶的菌株,并利用单因素和正交实验进行产酶条件优化,以提高其胶原蛋白酶活力。结果表明,8株耐高温、嗜中性和产胶原蛋白酶的菌株被选出。其中,胶原蛋白酶活力最...以透明圈法、Azocoll法和16S r DNA,分离及鉴定耐高温、嗜中性和产胶原蛋白酶的菌株,并利用单因素和正交实验进行产酶条件优化,以提高其胶原蛋白酶活力。结果表明,8株耐高温、嗜中性和产胶原蛋白酶的菌株被选出。其中,胶原蛋白酶活力最高的菌株鉴定为Bacillus glycinifermentans。对B.glycinifermentans的产酶条件进行优化,确定了其最佳产酶条件为培养温度50℃、p H8.0、培养时间16 h、种龄10 h、装液量100 m L和接种量5%。条件优化后使其胶原蛋白酶活力由1.0215 U/m L提高至(5.97±0.52)U/m L。成功筛选到的目标菌,经过条件优化使其最大酶活力提高了4.8倍,为后续鱼骨的高效发酵提供了基础。展开更多
The aim of this study was to find a way to efficiently separate neuronal cells from the cerebral cortex of adult rats,providing a reference method for rapid acquisition of neuronal cells from the adult rat brain.Fifte...The aim of this study was to find a way to efficiently separate neuronal cells from the cerebral cortex of adult rats,providing a reference method for rapid acquisition of neuronal cells from the adult rat brain.Fifteen SD rats were randomly divided into three groups,with five SD rats in each group.Then,neuron cells were isolated from the adult rat cerebral cortex by the grinding method,the trypsin method,and the collagenase II method,respectively.The expression of anti-NeuN in the neurons of each group was analyzed by flow cytometry.The acquisition rates and morphology of neurons of each group were observed by immunofluorescence staining.The grinding or collagenase II method is more suitable for rapid acquisition of neuronal cells from an adult rat’s cerebral cortex.The number of neuron cells obtained by the trypsin method were very few,so it is not convenient for later experiments.展开更多
基金the National Natural Science Foundation of China(No.81960129)Qinghai basic Research Plan Project(No.2019-ZJ-922)Middle-aged and Youth Foundation of Qinghai university affiliated hospital(No.2018-QYY-13).
文摘The aim of this study was to find a way to efficiently separate neuronal cells from the cerebral cortex of adult rats,providing a reference method for rapid acquisition of neuronal cells from the adult rat brain.Fifteen SD rats were randomly divided into three groups,with five SD rats in each group.Then,neuron cells were isolated from the adult rat cerebral cortex by the grinding method,the trypsin method,and the collagenase II method,respectively.The expression of anti-NeuN in the neurons of each group was analyzed by flow cytometry.The acquisition rates and morphology of neurons of each group were observed by immunofluorescence staining.The grinding or collagenase II method is more suitable for rapid acquisition of neuronal cells from an adult rat’s cerebral cortex.The number of neuron cells obtained by the trypsin method were very few,so it is not convenient for later experiments.
