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Mutant strain of synechocystis PCC6803 and application thereof

A technology of PCC6803, mutant strains, applied in application, microorganism-based methods, bacterial peptides, etc., can solve problems such as low tolerance, inhibition of cyanobacterial growth, and biomass decline, and achieve the effect of improving tolerance

Active Publication Date: 2017-11-28
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because ethanol is highly toxic to cyanobacteria, it will inhibit the growth of cyanobacteria, resulting in a decrease in their biomass, and ultimately lead to low ethanol production
It can be seen that the low tolerance of cyanobacteria to ethanol is the key factor affecting the use of cyanobacteria as a chassis to limit ethanol production

Method used

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  • Mutant strain of synechocystis PCC6803 and application thereof
  • Mutant strain of synechocystis PCC6803 and application thereof
  • Mutant strain of synechocystis PCC6803 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Construction of PCC 6803△slr0599 mutant strain:

[0020] 1) In vitro amplification of the target gene

[0021] Gene knockout is achieved through homologous recombination. In the present invention, the slr0599 gene is replaced with the kanamycin resistance gene, and the upstream and downstream gene fragments of slr0599 are used as upstream and downstream homologous arms for homologous recombination.

[0022] Using the CTAB method to extract the Synechocystis 6803 genome as a template, SEQ ID No.1 and SEQ ID No.2 are the upstream and downstream primers of the resistance gene respectively; SEQ ID No.3 and SEQ ID No.4 are respectively the primers to be knocked out The upstream and downstream primers of the upstream fragment of the gene slr0599; SEQ ID No.5 and SEQ ID No.6 are the upstream and downstream primers of the downstream fragment of the gene slr0599 to be knocked out respectively, and PCR amplification is carried out to obtain the Kana resistance gene, which is to b...

Embodiment 2

[0077] Analysis of Growth State of PCC 6803 △slr0599 Mutant Strain Under Ethanol Stress

[0078] Measure the whole-cell absorption spectrum of the mutant algal strain obtained in Synechocystis sp. PCC6803 wild-type and Example 1 under 1.5% (v / v) ethanol stress: the above-mentioned two kinds of algae grown to the logarithmic phase are respectively used as seed liquid, were inoculated into 250mL conical flasks containing 50mL of BG11 medium, and the initial OD of each bottle of algae 730 = 0.1. The culture conditions were 30°C, 150rpm, and 50μE·m for continuous culture for 4 days. -2 ·s -1 Continuous lighting. At the beginning of the culture, ethanol was added to the culture medium of the wild strain and the mutant strain respectively to a final concentration of 1.5% (v / v), and the total cell uptake was measured every 24 hours. The experimental group and the control group each had three parallels.

[0079] Take 3mL of the algae liquid cultured to the 4th day in the growth c...

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Abstract

Belonging to the field of industrial microorganisms, the invention in particular relates to a synechocystis PCC6803 mutant strain with significantly improved ethanol tolerance and application thereof. The ethanol tolerance related gene is slr0599, and the base sequence is shown as SEQ ID No.7. The mutant strain PCC 6803Deltaslr0599 is preserved on August 4, 2017 in China General Microbiological Culture Collection Center (CGMCC) located at No.3, 1st yard, Beichen west road, Chaoyang District, Beijing, the preservation number is CGMCC14333, and the strain is named as Synechocystis sp. by taxonomy. The synechocystis mutant strain is an ethanol tolerant mutant strain, and under 1.5% (v / v) ethanol stress, the growth state of the algal strain is obviously superior to that of wild algal strains. The ethanol tolerant algal strain obtained by the invention has important application value and referential significance for production of biofuel ethanol with synechocystis, and has wide application prospect.

Description

technical field [0001] The invention belongs to the field of industrial microorganisms, and in particular relates to a Synechocystis PCC6803 mutant strain with significantly improved ethanol tolerance and application thereof. Background technique [0002] With the emergence of problems such as the depletion of traditional energy sources, bioenergy has attracted more and more attention. Bioethanol is an excellent renewable energy source, which has the characteristics of less pollution and low price, and has broad development prospects. At present, the production of ethanol basically uses crops as raw materials directly. With the continuous increase of ethanol production and consumption, more and more crops are consumed, which will exacerbate the shortage of food and land resources in the world, and then trigger a series of economic and ethical issues. question. Second, the process of cultivating food crops for biofuel production requires large amounts of water, fertilizers ...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N1/21C12P7/06C12R1/01
CPCC07K14/195C12P7/065Y02E50/10
Inventor 赵佳琳崔玉琳王肖肖秦松
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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