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Heat-resistant halomonas bacterium and application thereof

A technology of halophilic bacteria and moderate halophilic bacteria, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., can solve problems such as pollution, non-degradable, and environmental damage

Inactive Publication Date: 2019-10-18
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] With the continuous development of social productivity and the improvement of people's living standards, the amount of plastics has greatly increased, but as far as the currently used plastics are concerned, they are all non-degradable, and many of them are disposable, causing serious damage to the environment and Pollution

Method used

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  • Heat-resistant halomonas bacterium and application thereof

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Experimental program
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Effect test

Embodiment 1

[0021] Embodiment one: Halomonas Halomonas Screening and isolation of sp. LCB169

[0022] The samples were collected from saline-alkali soil collected from Yumen Pass in Dunhuang, Gansu Province. Add 1 g of soil sample into a test tube filled with 9 mL of sterile water, vortex, then pipette 1 mL of the suspension into a test tube filled with 9 mL of sterile water, and prepare 10 -1 ~10 -6 Multiple times of the soil dilution suspension, pipette 100 μL and spread it on the screening medium plate, place it upside down at 37 °C and incubate for 72 hours, observe the growth status and morphological characteristics of the colony, pick a single colony with a sterile bamboo stick and transfer To fresh LB+10 % (w / v)NaCl (pH=8.0) solid medium for streak purification, cultivate under the same conditions, pick out a single colony and purify again, repeat several times, and finally pick a single colony for transfer To the slant of the purified medium, store at 4 °C for short-term stora...

Embodiment 2

[0026] Embodiment two: Halomonas Halomonas growth characteristics of sp. LCB169

[0027] Prepare LB liquid medium (pH=8.0) with NaCl concentration of 0%, 3%, 5%, 7%, 10%, 12%, 15%, 17% and 20% (w / v) respectively, each gradient Set up three parallels, quantitatively inoculate each test tube with fresh seed solution, place it on a shaker at 30°C and 180 rpm for 48 h, take it out and measure its OD 600 value, so as to determine the tolerance range and optimal growth concentration of its NaCl concentration; similarly, use the buffer system (pH 4.0–5.0: 0.1 M citric acid / 0.1 M sodiumcitrate; pH 6.0–8.0: 0.1 M KH 2 PO 4 / 0.1 M NaOH; pH 9.0–10.0: 0.1 M NaHCO 3 / 0.1 MNa 2 CO 3 ; pH 11.0: 0.05 M Na 2 HPO 4 / 0.1 M NaOH) to prepare LB+10 % (w / v) NaCl liquid medium with pH values ​​of 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, and analyze the pH tolerance range after inoculation and optimal growth pH; combined with the existing conditions of this experiment and comprehensively c...

Embodiment 3

[0028] Embodiment three: Halomonas Halomonas Analysis of enzymatic characteristics of sp. LCB169

[0029] Oxidase test: Add fresh reagent of 1 % (w / v) tetramethyl-p-phenylenediamine hydrochloride directly to a single colony in the exponential growth phase, if it turns purple within 10 s, it is oxidase Positive; on the contrary, if it changes color or does not change color for more than 60 s, it is oxidase negative.

[0030] Catalase experiment: the experimental strain was cultured in the solid medium of LB+10% (w / v) NaCl to the exponential growth phase, a little bacterium was picked and applied to a petri dish, and 5% hydrogen peroxide solution was added dropwise, Observe whether there are bubbles, if a large number of bubbles are produced within 30 s, it is catalase positive; otherwise, if no bubbles are produced, it is catalase negative.

[0031] Urease test: culture the experimental strains on LB+10% (w / v) NaCl medium plates for urease determination on the 3rd and 7th da...

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Abstract

The invention discloses a heat-resistant moderately halophilic bacterium halomonas sp. LCB169 of which the preservation number is KCTC 52618. The strain is determined as a new species of halomonas bya polyphasic classification method. The halomonas bacterium disclosed by the invention has enzyme activities of oxidase, catalase, esterase, lipoesterase, leucine aminopeptidase, valine aminopeptidase, alkaline phosphatase and the like, and can be widely applied in related production fields of various bioengineering enzyme preparations. Moreover, the strain can grow normally at an NaCl concentration up to 17% and high temperature of 52 DEG C, indicating that the strain has high resistance to the environment. At the same time, the strain can secrete extracellular polysaccharides and accumulatepolyhydroxybutyrate abundantly under high NaCl concentration of up to 10%, so the strain can be further developed and applied to biomedicine and environmentally-friendly bioplastics related industriessuitable for extreme high-salt environments.

Description

technical field [0001] The invention relates to the field of microbial strains and applications thereof, in particular, the invention relates to the technical field of thermotolerant halophilic monas and its applications. Background technique [0002] Extremophile microorganisms have become a hotspot of scientific research because of their special living environment and physiological mechanism. Halophilic bacteria are a special type of microorganisms that can only obtain the optimum growth state in a relatively high concentration of salt environment. However, the high-salt environment has obvious toxic effects on common microorganisms. The main reason is that the cell membrane and enzymes are destroyed under the stress of high osmotic pressure, thereby inhibiting the physiological activities of microorganisms. Due to their special physiological structure and metabolic regulation mechanism, halophilic bacteria can inhabit and reproduce in high-salt extreme environments. [...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/04C12R1/01
CPCC12P19/04C12N1/205C12R2001/01
Inventor 田永强甘龙站李晓广
Owner SICHUAN UNIV
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