Three newly-separated vibrio fast-growing strains and application thereof

A vibrio, rapid technology, applied in the direction of bacteria, microorganisms, microorganisms, etc., can solve the problems that cannot be applied, and there are few researches on the industrial application potential of vibrio

Active Publication Date: 2020-08-07
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 a

Method used

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  • Three newly-separated vibrio fast-growing strains and application thereof
  • Three newly-separated vibrio fast-growing strains and application thereof
  • Three newly-separated vibrio fast-growing strains and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0030] The steps are:

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

[0032] Step 2: Inoculate the bacterium solutions of the three activated strains into the new 5ml LB3 medium that has been sterilized in advance with an inoculation amount of 1%. Under the three temperature conditions of 30° C., 37° C. and 42° C., the cultivation was carried out at 200 rpm respectively.

[0033] After 5 hours, observe the turbidity of the bacteria in the shaking 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. Vibrio sp.FA1 strains were tested for growth with MKO and VN chemically synthesized media respectively

[0035] The steps are:

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

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

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

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

[0040] MKO medium: 20g gl...

Embodiment 3

[0043] Example 3: Vibrio sp.FA2 strains were tested for growth with MKO and VN chemically synthesized media

[0044] The steps are:

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

[0046] Step 2. The activated bacteria were transferred to sterilized 100ml MKO and VN culture medium with 1% inoculum, cultured at 37°C and 200rpm;

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

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

[0049] MKO medium: 20g glucose, 15g NaCl, 10g K 2 HPO 4 , 2g KH...

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Abstract

The invention discloses three newly-separated vibrio fast-growing strains and the application of the vibrio fast-growing strains and relates to the field of bioengineering. The three strains of vibrios are respectively named as Vibrio sp. FA1, Vibrio sp. FA2 and Vibrio sp. FA3, and the three strains of vibrio sp. FA2 and Vibrio sp. FA3 are respectively named as Vibrio sp. The strain is preserved in China Center for Type Culture Collection on August 5, 2019, the address is Wuhan University, China, and the preservation numbers are CCTCC NO: M 2019603, CCTCC NO: M 2019604 and CCTCC NO: M 2019605in sequence. Compared with the known Vibrio natriensis ATCC 14048 with the fastest growth speed, the three strains of vibrio natriensis ATCC 14048 have the obvious growth advantages that the vibrio natriensis ATCC 14048 has the highest growth speed; whole genome comparative analysis finds that more DNA replication-related genes, amino acid synthesis-related genes and stress resistance-related genes exist; the three vibrios can quickly grow by adopting various carbon sources under the condition of a chemical synthesis culture medium, and industrial application is facilitated. In addition, electro-transformation operation transformation plasmids and gene knockout prove the genetic operability of the strains, and important foundation and basis are provided for application of the three vibriosin the technical field of industry.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to three newly isolated fast-growing Vibrio strains and applications thereof. Background technique [0002] Vibrio is a rod-shaped, Gram-negative bacteria that can move through polar flagella and can carry out facultative anaerobic metabolism. At present, there are 8 effective genera in the family Vibrio, which are Vibrio, Enterovibrio, Halovibrio, Alternative Vibrio, Streptococcus, Glimonella and Echinomonas. Vibrio is widely distributed in the seawater of estuaries, bays and other sea areas and is abundant in seawater. Among them, there are more than ten species that may cause infection and disease in humans and various aquatic animals, such as Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus . [0003] Because Vibrio has a fast growth rate, a wide range of pH and can grow in seawater of various salinities, seawater can be directly used for fermentation. At the same ...

Claims

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Application Information

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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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