A kind of fast-growing vibrio and its application

A vibrio, rapid technology, applied in the field of bioengineering, can solve the problems of vibrio's industrial application potential, which is rarely studied and cannot be applied

Active Publication Date: 2022-07-26
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, most of the research on Vibrio is on the strain Vibrio natriegens ATCC 14048, and there are few studies on the industrial application potential of Vibrio
In addition, most studies have used rich media such as LB, which cannot be applied in large-scale fermentation

Method used

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  • A kind of fast-growing vibrio and its application
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1. Test of growth temperature of Vibrio sp.FA1, Vibrio sp.FA2, Vibrio sp.FA3

[0030] The steps are:

[0031]Step 1: Take the glycerol tubes of the three strains of Vibrio sp.FA1, Vibrio sp.FA2 and Vibrio sp.FA3 out of the -80°C refrigerator and inoculate them into 5ml LB3 medium (3% sodium chloride) with 1% inoculum each. , 1% peptone, 0.5% yeast powder), 37 ° C, 200 rpm overnight activation culture;

[0032] Step 2: The bacteria liquids of the three activated bacteria were respectively inoculated into a new 5ml LB3 medium that had been sterilized beforehand at an inoculation amount of 1%. Cultures were carried out at 200 rpm under three temperature conditions of 30°C, 37°C and 42°C, respectively.

[0033] After 5 hours, observe the degree of turbidity of the bacteria in the shake tube at this time, and measure the OD of the culture at this time. 600 It was found that the three strains could grow normally under the three temperature conditions.

Embodiment 2

[0034] Embodiment 2, carry out growth test to Vibrio sp.FA1 strain with MKO and VN two kinds of chemical synthesis medium respectively

[0035] The steps are:

[0036] Step 1. Take the glycerol tube of Vibrio sp.FA1 out of the -80°C refrigerator and inoculate 5ml LB3 medium (3% sodium chloride, 1% peptone, 0.5% yeast powder) with 1% inoculum at 37°C, 200rpm overnight activation culture;

[0037] Step 2: Transfer the activated bacteria to sterilized 100ml MKO and VN medium with 1% inoculum respectively, and cultivate at 37°C and 200rpm;

[0038] Step 3. Measure OD with a spectrophotometer every two hours 600 The value of SBA-40D biosensing analyzer (purchased from Shandong Academy of Sciences, Jinan, China) was used to measure the amount of remaining glucose in the medium to record the growth and sugar consumption of bacteria.

[0039] The formulations of the MKO and VN culture medium used in this example are as follows:

[0040] MKO medium: 20g glucose, 15g NaCl, 10g K 2 ...

Embodiment 3

[0043] Embodiment three, carry out growth test to Vibrio sp.FA2 strain with MKO and VN two kinds of chemical synthesis medium respectively

[0044] The steps are:

[0045] Step 1. Take the glycerol tube of Vibrio sp.FA2 out of the -80°C refrigerator and inoculate 5ml LB3 medium (3% sodium chloride, 1% peptone, 0.5% yeast powder) with 1% inoculum at 37°C, 200rpm overnight culture;

[0046] In step 2, the activated bacteria were transferred to sterilized 100ml MKO and VN medium with 1% inoculum respectively, and cultivated at 37°C and 200rpm;

[0047] Step 3. Measure OD with a spectrophotometer every two hours 600 The value of SBA-40D biosensing analyzer (purchased from Shandong Academy of Sciences, Jinan, China) was used to measure the amount of remaining glucose in the medium to record the growth and sugar consumption of bacteria.

[0048] The formulations of the MKO and VN culture medium used in this example are as follows:

[0049] MKO medium: 20g glucose, 15g NaCl, 10g ...

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Abstract

The invention discloses three newly isolated fast-growing Vibrio strains and applications thereof, and relates to the field of bioengineering. The three Vibrio strains are named Vibrio sp.FA1, Vibrio sp.FA2 and Vibrio sp.FA3 respectively, and were published on August 2019. It was deposited on the 5th in the China Center for Type Culture Collection, address: Wuhan University, China, and the deposit numbers are CCTCC NO: M 2019603, CCTCC NO: M 2019604, CCTCC NO: M 2019605. Compared with the known fastest growing Vibrio natriegens ATCC 14048, these three Vibrio strains have obvious growth advantages. Through genome-wide comparative analysis, it is found that they also have more DNA replication-related genes, amino acid synthesis-related genes and resistance Inversion-related genes; these three Vibrio strains can grow rapidly using a variety of carbon sources under the condition of chemically synthesized medium, which is more conducive to industrial application. In addition, the transformation plasmids and gene knockout by electrotransformation proved the genetic operability of these strains, which provided an important basis and basis for the application of these three strains of Vibrio in the field of industrial technology.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a fast-growing vibrio and its application. Background technique [0002] Vibrio is a class of rod-shaped, gram-negative bacteria that can move through polar flagella and undergo facultative anaerobic metabolism. At present, the Vibrio family contains 8 effective genera, namely Vibrio, Enterovibrio, Halovibrio, Altvibrio, Streptococcus, Grimmonella and Echinomonas. Vibrio is widely distributed in sea waters such as estuaries and bays and is abundant. Among them, there are more than ten species that may cause human and various aquatic animal infections and diseases, such as Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus. . [0003] Because of the fast growth rate of Vibrio, the wide range of pH growth, and the ability to grow in seawater of various salinities, seawater can be directly used for fermentation. At the same time, Vibrio has rich physiological and biochem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/74C12R1/63
CPCC12N15/74C12R2001/63C12N1/205
Inventor 陶飞彭源许平唐鸿志
Owner SHANGHAI JIAO TONG UNIV
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