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Cyanobacteria strain capable of being used for glycosylglycerol production and application thereof

A technology of glucoside and cyanobacteria, which is applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of field environment tolerance and scale cultivation technology limitations of industrial application production cost strains, and achieve strong environmental adaptability, The effect of strong synthetic ability

Active Publication Date: 2018-12-11
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In summary, the current research on the production of glycerol glucoside by model cyanobacteria has proved the feasibility of the technical route, but its industrial application is still limited by factors such as production cost, strain field environmental tolerance, and large-scale cultivation technology.

Method used

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  • Cyanobacteria strain capable of being used for glycosylglycerol production and application thereof
  • Cyanobacteria strain capable of being used for glycosylglycerol production and application thereof
  • Cyanobacteria strain capable of being used for glycosylglycerol production and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Morphological identification of algal strains

[0027] 1. Optical microscope observation method

[0028] 1) Turn on the power of the optical microscope (BX51, olympus, Japan), and adjust the condenser to the appropriate brightness of the field of view.

[0029] 2) Will grow to OD 730 =1.0 (about 4 days of culture) took 1 drop of the algae solution, dropped it on a clean glass slide, and covered it with a cover glass.

[0030] 3) First find the cells of the algae strain under the 40 times objective lens, then drop 1 drop of cedar oil on the cover glass, observe the cell morphology of the algae strain under the 100 times objective lens, and take microscopic photos of the cells (see figure 1 ).

[0031] 2. SEM sample preparation method

[0032]1) Sampling: take 15mL1B1 algae liquid (OD 730 =1.0-1.5) centrifuge at 4000rpm for 1min, remove the supernatant, add 1mL of PBS phosphate buffered saline (8g L -1 NaCl, 0.24gL -1 K H 2 PO 4 , 0.2g L -1 KCl, 1.44...

Embodiment 2

[0051] Embodiment 2: Algae strain species identification based on 16S ribosomal DNA sequence

[0052] 1. Amplification and sequencing of 16S ribosomal rDNA sequence of 1B1 strain

[0053] Take 1mL 1B1 algae cell culture (OD 730 =1.0), the cells were collected by centrifugation; resuspended in 20 μL of sterile water, and repeatedly frozen and thawed in liquid nitrogen and 65° C. for 6 times. Centrifuge at 8000 rpm for 1 minute. 1.5 μL of the supernatant was used as a template for the PCR reaction, and the 16S rDNA fragment was amplified by PCR using sgF / sgR as the primer pair (the components and procedures of the PCR reaction are shown in Tables 1 and 2, respectively). The obtained PCR fragments were directly sent to the sequencing company for determination, and the DNA sequences shown in Sequence Table 1 were obtained.

[0054] Table 1 PCR reaction system

[0055]

[0056] Table 2 PCR reaction program

[0057]

[0058] Primers used in Table 3

[0059]

[0060] 2...

Embodiment 3

[0079] Example 3: Evaluation of the salt tolerance, growth rate, and synthetic ability of compatible substances of 1B1 algae strain

[0080] 1. Evaluation of the salt tolerance of algal strains

[0081] Inoculate 1B1 strain and Synechocystis PCC6803 strain into BG11 medium containing 0mM, 300mM, 600mM and 900mM sodium chloride, respectively, at 30°C, 30μE / m 2 / s light intensity, cultivated on a shaker at 150rpm for one week, and observed the growth of algal strains. The composition of BG11 medium is 1.5g L -1 NaNO 3 ,40mgL -1 K 2 HPO 4 ·3H 2 O, 36 mg L -1 CaCl 2 2H 2 O, 6 mg L -1 Citric acid, 6mg L -1 Ammonium ferric citrate, 1mg L -1 EDTA disodium salt, 20mg L -1 NaCO 3 ,2.9mg L -1 h 3 BO 3 ,1.8mgL - 1 MnCl 2 4H 2 O, 0.22 mg L -1 ZnSO 4 ·7H 2 O, 0.39 mg L -1 NaMoO 4 2H 2 O, 0.079 mg L -1 CuSO 4 ·5H 2 O and 0.01 mg L -1 CoCl 2 ·6H 2 O.

[0082] 2. Evaluation of growth rate of algal strains

[0083] Insert 1B1 strain and Synechocystis PCC6803 ...

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Abstract

The invention relates to the field of synthesis of high additional value chemicals by microorganisms, in particular to a cyanobacteria strain capable of being used for glycosylglycerol production andan application thereof. The strain is cyanobacterium aponinum 1B1 which is preserved in CGMCC on the 2nd, March, 2017, the preservation number of CGMCC No. 13785. The alga strain screened is separatedfrom a natural ecological environment of a salt lake in Yuncheng, Shanxi. Compared with a mode cyanobacteria synechocystis PCC6803, the strain has higher GG synthetic ability and adaptive capacity toenvironment. A compatible matter synthesized by the alga strain under salt stress is glycosylglycerol, so that the product is separated favorably. Therefore, the strain is quite suitable for being used as a production strain for glycosylglycerol and is applied to industrial production.

Description

technical field [0001] The invention relates to the field of microbial synthesis of high value-added chemicals. Specifically, a cyanobacterial strain producing glycerol glucoside and its application are provided. Background technique [0002] Cyanobacteria (cyanobacteria) are prokaryotic microorganisms capable of plant-type oxygen-evolving photosynthesis and are important primary producers in marine ecosystems (Hernandez-Prieto, Semeniuk et al. 2014). Compared with higher plants, cyanobacteria have the advantages of fast growth, high photosynthetic efficiency, and easy genetic modification (Zhou and Li 2010). Alternatively, aquatic cyanobacteria can be grown in incubators placed on inhospitable land or even floating in the ocean. Based on these advantages, cyanobacteria have demonstrated their great potential in the production of biofuels, bio-based chemicals and carbohydrates, and have broad application prospects (Gupta, Ratha et al.2013, Angermayr, Rovira et al.2015, Ha...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/44C12P19/04C12R1/01
CPCC12P19/04C12P19/44C12N1/205C12R2001/01
Inventor 吕雪峰钟成伟谈晓明
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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