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Mechanism of morphological development of mycelia and its application on production of microbial product

菌丝形态发育机制及其在微生物产品生产中的应用

基本信息

批准号：
13650854
负责人：
PARK Enoch y.
金额：
$ 2.3万
依托单位：
SHIZUOKA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13650854/
关键词：
Image analysis Flow through chamber Mycelial morphology Arachidonic acid Mortierella alpina Fungus

项目摘要

The intrastructure of mycelial pellets of Mortierella alpina and the morphological parameters, such as hyphal growth rate, tip formation rate, tip extension rate and branch formation rate of Mortierella alpina have been investigated in this research. The intrastructure of mycelial pellets were visualized following labeling with fluorescein isothiocyanate (FITC) and Nile red using fluorescence microscopy. The pellet was an ellipse shape, but its intrastructure was shaped as doughnut and a cave was present inside, which was clear by judging 3-D image of intrastructure of pellet. Using image analysis, the fatty acid production was shown on the verge of pellet, which was corresponded on the place of dense mycelia. The cavity ratio was calculated from area based on the fluorescent intensity and remained at 20% in a 50-L fermentor but finally increased to 35%. Mycelial pellet volume was paralleled to the cavity ratio.Using a flow-through chamber under 25 different combinations of carbon and nitrogen concentrations, morphological parameters were measured and found that they were influenced not by C/N ratio but by carbon concentration in medium. Specific rates of hyphal growth and tip formation remained at constant at lower carbon concentration of 5 g/l, respectively. Tip extension rate from one tip was 60 (μm/tip/h) in a carbon concentration below 15 g/l, and branching formation rate was independent upon carbon concentration. Tip extension rate was a function of specific hyphal growth rate, and the specific hyphal growth rate was linearly proportional to the specific tip formation rate, which demonstrated that tip extension rate was exponentially proportional to the specific tip formation rate. Branch formation rate per hyphal element remained unchanged even at lower tip extension rate than 60 (μm/tip/h) and at lower specific hyphal growth rate than 0.83 h^<-1>, respectively.

本研究对高山孢霉菌丝球的基本结构和菌丝生长速率、尖端形成速率、尖端延伸速率和分支形成速率等形态学参数进行了研究。用荧光显微镜观察异硫氰酸荧光素（FITC）和尼罗红标记菌丝球的结构。颗粒为椭圆形，但其基础结构呈甜甜圈状，内部存在洞穴，从颗粒基础结构的三维图像可以看出这一点。通过图像分析，脂肪酸的产生在球团的边缘，与致密菌丝体的位置相对应。空腔比根据荧光强度从面积计算，在50-L发酵罐中保持20%，最终增加到35%。菌丝球体积与空腔比平行。在25种不同的碳氮浓度组合下，利用流动室测量了形态参数，发现它们不受碳氮比的影响，而是受介质中碳浓度的影响。在较低碳浓度为5 g/l时，菌丝生长速率和尖端形成速率保持不变。碳浓度低于15 g/l时，单根枝尖延伸速率为60 μm/枝尖/h，分支形成速率与碳浓度无关。叶尖延伸率是菌丝比生长率的函数，菌丝比生长率与叶尖比形成率成线性关系，说明叶尖延伸率与叶尖比形成率成指数关系。即使在茎尖延伸速率低于60 μm/茎尖/h和菌丝特定生长率低于0.83 h^<-1>时，每个菌丝元素的分枝形成速率保持不变。