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Culture medium for microvesicle bacteria BS03 and preparation method thereof

A technology of microvesicles and culture medium, applied in the field of culture medium and its preparation of microvesicles BS03, which can solve the problems of long test period, failure to display the interaction relationship of various factors, and heavy workload

Inactive Publication Date: 2011-08-17
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Traditional optimization methods such as the single-factor method to optimize the medium can only consider the influence of one factor level at a time, which requires a large workload, multiple experiments and a long test cycle, and cannot show the interaction between the various factors.

Method used

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  • Culture medium for microvesicle bacteria BS03 and preparation method thereof
  • Culture medium for microvesicle bacteria BS03 and preparation method thereof
  • Culture medium for microvesicle bacteria BS03 and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Using 2216E as the basal medium, the single factor method was used to optimize the culture conditions.

[0030] The influence of temperature: Set the culture temperature as 20, 25, 28, 30 and 35°C respectively, pH 7.0-7.2, 1% inoculum size, 50ml liquid volume in 250ml triangular shaker flask, 180r / min shaking table after 24h cultivation, Bacterial solution was taken to measure OD 600 and LD 50 .

[0031] Influence of rotation speed: Set the shaking speed of the shaker to 120, 150, 180 and 210r / min respectively, and the rest of the conditions are the same as above. After culturing for 24 hours, take the fermentation broth to measure the OD 600 and LD 50 .

[0032] The influence of initial pH: choose 2.0mol / l HCl and 2.0mol / l NaOH respectively to adjust the pH of the medium to 5.0, 6.0, 7.0, 8.0 and 9.0, the temperature is 28°C, the inoculum size is 1%, and the liquid in the 250ml Erlenmeyer shaker flask The volume is 50ml, after 24 hours of culture on a shaker at 18...

Embodiment 2

[0037] In this embodiment, several composite nutrient sources are firstly selected, that is, those nutrients that can be used as both carbon source and nitrogen source (such as peptone, yeast powder, etc.) and two kinds of inorganic nitrogen sources. Then on their basis, add different micro-components to investigate the effects of each component on the growth of microvesicles and the production of active substances (see Figure 1~5 ).

[0038] With 2216E as the basic medium (without carbon and nitrogen sources), 5.0g / L soybean peptone, peptone, beef extract, yeast powder were used as the compound nutrient source and NaNO 3 and KNO 3 as an inorganic nitrogen source. The content of other components is the same as that of the basal medium. The culture conditions are: 1% inoculum size, 28°C, 180r / min, pH 7, salinity 30‰, measure OD after shaking the flask for 24 hours 600 and LD 50 . The results showed that the BS03 strain was more suitable for growing in organic nitrogen. ...

Embodiment 3

[0044] According to the previous experiments, peptone, sucrose, pH, inoculum size, and culture time were selected as the influencing factors for a uniform design of 5 factors and 15 levels, and the rest of the components were the same as the initial medium. The five factors are independent variables X1, X2, X3, X4 and X5, dry weight of bacteria and OD 600 is the dependent variable Y 1 , Y 2 . Selection of culture conditions: 28°C, 180r / min, salinity 30‰. Each factor is set at 15 levels, and the experimental results are subjected to the quadratic polynomial stepwise regression analysis of the DPS software data processing system, and the model is tested for significance (see Table 1, Table 2)

[0045] Table 1

[0046]

[0047] The regression equation is:

[0048] Y=5.38134906+113.60189202X1×X1-3.793014914X2×X2+2.5854730603X3×X3-28.422411492X4×X4-0.15092230997X5×X5-65.98236457X1×X3-270.25223629X1×X5-10.298424271X2×X3+20.758363788X2×X4+49.38660676 X2×X5+28.185002912X3×X4+...

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Abstract

The invention relates to a culture medium for microvesicle bacteria BS03 and a preparation method thereof, relating to a microbial culture medium and providing a culture medium, which is suitable for efficient culture of the microvesicle bacteria BS03 and can obviously increase cell live weight of the microvesicle bacteria BS03 and yield of algae-killing active substances, and a preparation method of the culture medium. The culture medium comprises 1L of distilled water, 1-20g / L of peptone and 1-15g / L of cane sugar. The preparation method of the culture medium comprises the steps of: adding the peptone and the cane sugar into the distilled water, enabling a constant volume to be 1L, and sterilizing to obtain the culture medium of the microvesicle bacteria BS03. The composite culture medium suitable for high-dense culture of the microvesicle bacteria BS03 is developed by taking 2216E as a basic culture medium and adopting various methods for optimizing culturing conditions.

Description

technical field [0001] The invention relates to a microorganism culture medium, in particular to a culture medium of Microvesicle BS03 and a preparation method thereof. Background technique [0002] In recent years, due to the severe pollution and eutrophication of coastal waters, the frequency of red tides has increased sharply, and the scale has continued to expand. New red tide algae species continue to appear, and the proportion of toxic red tide algae species has increased. It occurs frequently in more than 30 countries and regions including the United States, Japan, China, Canada, France, Sweden, Norway, the Philippines, India, Indonesia, Malaysia, South Korea, and Hong Kong. To reduce the degree of eutrophication of water bodies, it is imperative to seek effective ways to prevent and control red tide and its toxin pollution. Red tide control has become an urgent problem to be solved at home and abroad. [0003] At present, algae control techniques can be classified i...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
Inventor 郑天凌傅丽君田蕴李东安新丽陈勇景晓明
Owner XIAMEN UNIV
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