M-01解淀粉芽孢杆菌的分离纯化鉴定及其生态培养生长规律的研究
发布时间：2019-02-10 19:45
【摘要】：本研究对从东北地区自行采集的菌株进行了分离、纯化、培育,并进行了相关的分子生物学技术鉴定和相关酶活力变化规律的研究。利用自行研制的智控益生菌液体发酵器和智控固体生态培养.干燥一体机对菌株进行了液体发酵与固体生态培养及干燥一体化研究,探寻了其相关指标动态变化规律,测试智控固体生态培养.干燥一体机的运行效果,为其工厂规模化生产应用提供了必要的技术参考,具有较好的理论应用价值。研究结果如下：(1)对自行采集分离纯化的M-01菌株进行了形态学研究和生理生化测定,并采用16SrDNA测序以及gyrA基因序列系统发育树分析技术对菌株进行了分子生物学鉴定,鉴定结果：M-01菌株为解淀粉芽孢杆菌(M-01, Bacillus amyloliquefaciens subsp.plantarum),属于好氧菌。(2)在18℃、pH 7.2条件下,对M-01菌株进行分泌淀粉酶、蛋白酶、脂肪酶酶活力动态变化规律的研究,结果表明：M-01菌淀粉酶活力68h时,达到较高水平13.02U/dL；蛋白酶活力在56h后趋于稳定,至74h时为8.04U/dL：脂肪酶活力在56h时达到峰值,为1.159U/L.(3)利用自行研制的智控益生菌液体发酵器对M-01菌进行液体发酵培养,在适宜的液体发酵营养底物组成和接种量条件下,合理有效控制发酵液温度35±0.2℃,初始pH值7.2±0.1以及气体搅拌方式等发酵条件,结果表明：M-01菌经24h液体发酵,活菌数为146.675±4.05×108CFU/m L,芽孢率为91.76±1.50%；在整个发酵过程中,发酵液pH值呈下降趋势,发酵液中还原糖含量呈下降趋势,至24h时,还原糖含量为5.857±0.314mg/mL;在发酵前期0-4h,发酵液中氨基氮含量增高,4-24h期间,发酵液中氨基氮含量逐渐降低,与此同时,在整个发酵过程中微生物总蛋白(MCP)含量呈现持续上升趋势。(4)通过L9(33)正交试验设计,研究培养温度、菌料湿度、作用时间3因素对固体生态培养28h后的M-01菌芽孢形成率的影响,结果表明：各因素对M-01菌芽孢形成率影响的主次顺序为：培养温度菌料湿度培养时间。故在M-01菌固体发酵培养28h后,促进芽孢率形成的影响因素的最佳组合为A1B3C2,即培养温度45℃,菌料湿度55%,培养时间2h。(5)采用自行研制的智控固体生态培养.干燥一体机对M-01菌进行固体生态培养及温敏干燥效果的研究,自动控制发酵温度35℃±1.0℃,菌料湿度55%±5%,并调节菌料初始pH 7.2±0.1,根据正交设计结果,采用分阶段固体发酵培养,第一阶段0-28h固体培养,控制发酵温度35℃±1.0℃,菌料湿度55%±5%,发酵培养28h,以获得较高的活菌数；第二阶段28-30h固体培养,提高发酵温度至45℃,维持培养2h,以刺激芽孢的形成,以获得较高芽孢数。结果表明：至30h时,菌质中活菌数为224.25±4.47×108CFU/g,芽孢数为203.63±4.51×108CFU/g,芽孢形成率为90.8±0.73%。在自行研制的智控固体生态培养.干燥一体机中,培养终止后即刻进行43℃±2℃温敏脱水干燥,能有效保持菌体活性并能同时达到干燥效果,检测到干燥菌料中芽孢数210.05±4.96×1O8CFU/g,含水率5.53±0.12%,杂菌率0.681±0.003%。
[Abstract]:In this study, the self-collected strains of the Northeast China were isolated, purified and cultivated, and the related molecular biological identification and the change of the activity of the related enzymes were carried out. The self-developed intelligent control probiotics liquid fermentation device and the intelligent control solid ecological culture are utilized. The dry all-in-one machine has carried on the research of liquid fermentation and solid ecological culture and dry integration of the strain, and the dynamic changes of relevant indexes of the strain are explored, and the intelligent control solid ecological culture is tested. the operation effect of the drying all-in-one machine provides the necessary technical reference for the large-scale production and application of the factory, and has better theoretical application value. The results of the study were as follows: (1) The M-01 strain isolated and purified by self-collection was determined by morphological study and physiological and biochemical test, and the sequence of 16SrDNA and the phylogenetic tree of gyrA gene were used to identify and identify the strain. The M-01 strain is Bacillus amyloliquefaciens (M-01, Bacillus amyloliquefaciens subsp.planarum), and belongs to aerobic bacteria. (2) The activity of amylase, protease and lipase in M-01 strain was studied under the conditions of 18 鈩，
本文编号：2419483
[Abstract]:In this study, the self-collected strains of the Northeast China were isolated, purified and cultivated, and the related molecular biological identification and the change of the activity of the related enzymes were carried out. The self-developed intelligent control probiotics liquid fermentation device and the intelligent control solid ecological culture are utilized. The dry all-in-one machine has carried on the research of liquid fermentation and solid ecological culture and dry integration of the strain, and the dynamic changes of relevant indexes of the strain are explored, and the intelligent control solid ecological culture is tested. the operation effect of the drying all-in-one machine provides the necessary technical reference for the large-scale production and application of the factory, and has better theoretical application value. The results of the study were as follows: (1) The M-01 strain isolated and purified by self-collection was determined by morphological study and physiological and biochemical test, and the sequence of 16SrDNA and the phylogenetic tree of gyrA gene were used to identify and identify the strain. The M-01 strain is Bacillus amyloliquefaciens (M-01, Bacillus amyloliquefaciens subsp.planarum), and belongs to aerobic bacteria. (2) The activity of amylase, protease and lipase in M-01 strain was studied under the conditions of 18 鈩，
本文编号：2419483
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