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Method for preparing bacterium-algae cells through two-wheel coculturing and flocculation method

A technology of co-cultivation and chrysalis, which is applied in the field of preparing bacteria and algae cells by two rounds of co-cultivation and flocculation, can solve the problems of increasing the dry weight of plants, so as to increase growth rate and biomass, improve harvesting efficiency, and improve The effect of flocculation performance

Inactive Publication Date: 2016-06-22
ZHEJIANG GONGSHANG UNIVERSITY
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  • Abstract
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Problems solved by technology

Rice plants contain many endophytic microorganisms, which are important endophytic bacteria resources. Among them, Pantoea is a common endophyte in rice, which can promote plant growth and increase plant dry weight [Sánchez-Matamoros RC, et al.Structure of the O-antigen of the lipopolysaccharide isolated from Pantoea ananatis AEP17, arhizobacterium associated with rice.CarbohydrateResearch,2013,369:25-30.], the main mechanism of rice endophytic Pantoea to promote the growth of host rice includes secretion of auxin, promotion of nitrogen fixation and phosphorus dissolution, etc. (RuizaD, et al.. 49(6):902-912), because microalgae have both the photosynthesis of plant cells and the rapid growth of microbial cells, theoretically endophytes that have symbiotic functions on plants may also exert similar biological effects on microalgal cells In this regard, a small amount of research evidence has shown that microorganisms that have a growth-promoting effect on plants can also increase the biomass of microalgae (KimBH, et al. However, so far, there has been no research on the development of new technologies for the production of chlorella using the symbiosis of rice endophytes and microalgae

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  • Method for preparing bacterium-algae cells through two-wheel coculturing and flocculation method

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Embodiment 1

[0029] 1) The rice endophytic Pantoea strains were purchased from the China Agricultural Microbiology Collection Center (ACCC), and the code is 10454. After the strains were activated by conventional methods, they were inoculated into a Erlenmeyer flask containing 10mL of LB liquid medium for expansion. For expansion culture, the bacteria liquid in the logarithmic growth phase after the expansion culture for 16 hours is inoculated according to the volume ratio of bacteria liquid: liquid LB medium = 1:10, and then the second expansion culture is carried out. The culture conditions are 37°C, 200rpm constant temperature shaker dark culture. The second expansion culture 10h to OD 600 =1 is used as the seed cell of the co-cultivation of bacteria and algae. According to the growth curve, the growth of the bacteria is in the logarithmic phase. The effective number of viable bacteria (CFU, Colony-FormingUnits) in the culture solution is measured by the conventional plate counting meth...

Embodiment 2

[0035] 1) The rice endophytic Pantoea strains were purchased from the China Agricultural Microbiology Collection Center (ACCC), and the code is 10454. After the strains were activated by conventional methods, they were inoculated into a Erlenmeyer flask containing 10mL of LB liquid medium for expansion. For expansion culture, the bacteria liquid in the logarithmic growth phase after the expansion culture for 16 hours is inoculated according to the volume ratio of bacteria liquid: liquid LB medium = 1:10, and then the second expansion culture is carried out. The culture conditions are 37°C, 200rpm constant temperature shaker dark culture. The second expansion culture 10h to OD 600 =1 is used as the seed cell of the co-cultivation of bacteria and algae. According to the growth curve, the growth of the bacteria is in the logarithmic phase. The effective number of viable bacteria (CFU, Colony-FormingUnits) in the culture solution is measured by the conventional plate counting meth...

Embodiment 3

[0041] 1) The rice endophytic Pantoea strains were purchased from the China Agricultural Microbiology Collection Center (ACCC), and the code is 10454. After the strains were activated by conventional methods, they were inoculated into a Erlenmeyer flask containing 10mL of LB liquid medium for expansion. For expansion culture, the bacteria liquid in the logarithmic growth phase after the expansion culture for 16 hours is inoculated according to the volume ratio of bacteria liquid: liquid LB medium = 1:10, and then the second expansion culture is carried out. The culture conditions are 37°C, 200rpm constant temperature shaker dark culture. Continuous expansion was carried out three times, and the culture conditions were 37°C, 200rpm constant temperature shaker and dark culture. The third expansion culture 10h to OD 600 =1 is used as the seed cell of the co-cultivation of bacteria and algae. According to the growth curve, the growth of the bacteria is in the logarithmic phase. T...

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Abstract

The invention discloses a method for preparing bacterium-algae cells through a two-wheel coculturing and flocculation method. The method comprises the steps of 1, conducting the first cycle of coculturing on rice endogenous pantoea which is added for the first time and chlorella under illumination, so that after culturing is ended, a bacterium-algae coculturing solution of the first cycle is obtained; 2, supplementing rice endogenous pantoea which is added for the second time into the bacterium-algae coculturing solution of the first cycle, conducting the second cycle of bacterium-algae coculturing under illumination, so that after culturing is ended, a bacterium-algae coculturing solution of the second cycle is obtained; 3, conducting flocculation collection treatment on the bacterium-algae coculturing solution of the second cycle, so that the bacterium-algae cells are obtained. According to the method, bacterium-algae coculturing is conducted through the rice endogenous pantoea and the chlorella, the growth rate of the algae cells is raised, biomass of the algae cells is increased, meanwhile, flocculation performance of the algae cells is improved, application of chemical flocculating agents and toxic harm to the algae cells are reduced, algae cell collection efficiency is improved, algae cell collection cost is lowered, and thereby industrial production and development and application cost of chlorella are remarkably lowered.

Description

technical field [0001] The invention relates to the technical field of co-cultivation of bacteria and algae, in particular to a method for preparing bacteria and algae cells by two rounds of co-cultivation and flocculation. Background technique [0002] Chlorella is a single-celled green algae, which has strong environmental adaptability and is easy to cultivate on a large scale. The cells contain rich nutrients and are widely used in health food, feed, food additives, finely processed products and as raw materials for pharmaceutical preparations . Chlorella is rich in short-chain fatty acids and is a high-quality biodiesel raw material (Xu Jin, Xu Xudong, Fang Xiantao, Hu Hanhua. Screening of high oil-yielding chlorella and oil analysis. Acta Hydrobiology, 2012,36(3):426 -432.). Chlorella pyrenoidosa is a common chlorella species in my country. It has high protein content and rich essential amino acids. It is an excellent protein source for feed (Li Guoping. Analysis of a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/12C12R1/89
CPCC12N1/12C12N1/20
Inventor 赵艳
Owner ZHEJIANG GONGSHANG UNIVERSITY
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