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Method for culturing microalgae by adopting lignocellulose

A lignocellulose and lignin technology, applied in the direction of using microorganisms, single-cell algae, bacteria, etc., can solve the problems of high cost of carbon sources, achieve cheap raw material sources, wide range of raw material sources, and reduce the cost of enzymes

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

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Problems solved by technology

[0009] Aiming at the problem of high cost of carbon source in heterotrophic fermentation of microalgae in the prior art, the present invention provides a process using lignocellulosic agricultural and forestry wastes as raw materials. Cellulase preparations, thereby reducing the carbon source cost of microalgae heterotrophic fermentation

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  • Method for culturing microalgae by adopting lignocellulose
  • Method for culturing microalgae by adopting lignocellulose

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Embodiment 1: By the method of indirect connection, construct the cellulase preparation based on Clostridium thermocellum cellulite

[0051] Through seamless cloning, the tdk expression cassette (including the promoter of the gapDH gene) and the pyrF expression cassette (including the pyrF self-promoter) were cloned into the plasmid pHK (Mohr, G., Hong, W., Zhang, J., Cui, G.-Z., Yang, Y., Cui, Q., et al. (2013) A targetron system for gene targeting in thermophiles and its application in Clostridium thermocellum, PLoS One 8:e69032.) downstream of the antibiotic gene cat, And through the design of primers, NheI and XbaI restriction sites were added between tdk and pyrF expression cassettes, and EagI and MluI restriction sites were added downstream of pyrF for cloning of homology arm fragments, thereby constructing pHK-HR plasmid.

[0052] The cellulase Cel9K (exocellulase, encoded by the nucleic acid sequence 2113813 to 2111293 in the genome CP002416.1) in the celluloso...

Embodiment 2

[0058] Embodiment 2: By the method of indirect connection, construct the cellulase preparation based on Clostridium thermocellum cellulite

[0059] The difference from Example 1 is that the polypeptide fragment II or polypeptide fragment I is linked to the 3' end of cellulosome endonuclease CelZ (SEQ ID NO: 15). The constructed recombinant strain is cultured to the mid-logarithmic phase in GS-2 medium with 5 grams per liter of cellulose or cellobiose as the carbon source, and can be used as a whole-bacteria enzyme preparation for biosaccharification of lignocellulose.

Embodiment 3

[0060] Embodiment 3: By the method of direct connection, construct the cellulase preparation based on Clostridium thermocellum cellulosome

[0061] Using the overlap extension polymerase chain reaction method, cellulosic exonuclease Cel9-48 (SEQ ID NO: 16) was combined with the sequence of type II adhesion module CohIIct (SEQ ID NO: 7) or type I of Clostridium thermocellum The sequence of the docking module DocIct (SEQID NO:6) was directly connected, wherein the sequence of CohIIct or DocIct was connected to the 3' end of the Cel9-48 sequence to obtain the sequence of Cel9-48-DocIct or Cel9-48-CohIIct.

[0062] Using the Cel9-48-DocIct or Cel9-48-CohIIct sequence as the target sequence, using the MluI and EagI restriction sites to clone into the homologous recombination plasmid pHK-HR, using the lactate dehydrogenase gene clo1313_1160 as the targeted replacement sequence, The homologous recombination plasmid pHK-HR-cel9-48 was constructed. The upstream homology arm HR-up is t...

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Abstract

The invention provides a technology taking lignocellulose type agricultural and forest wastes as raw materials, aiming at the problem in the prior art that the cost of a carbon source in a microalgaeheterotrophic fermentation process. The technology comprises the steps of pre-treating lignocellulose raw materials, saccharifying, carrying out microalgae heterotrophic fermentation, carrying out post-treatment and the like. The technology adopts a strategy combining biological saccharifying of lignocellulose and the microalgae heterotrophic fermentation, so that the production cost of the microalgae heterotrophic fermentation is remarkably reduced, and the comprehensive utilization problem of the agricultural and forest wastes is also solved. Meanwhile, a cellulose preparation for catalyzingthe lignocellulose to be saccharified is adopted, so that the enzyme utilization cost of a saccharifying phase is remarkably reduced. Furthermore, a culture medium at a lignocellulose saccharifying phase and a fermentation culture medium in the technology can be circularly utilized, so that water can be remarkably reduced and the dosage of chemicals also can be remarkably reduced; the technologyhas great significance on industrialization.

Description

technical field [0001] The invention relates to the field of microbial fermentation engineering, in particular to a method for fermenting and cultivating microalgae using lignocellulose raw materials. Background technique [0002] As a new type of aquatic biomass resource, microalgae has the characteristics of short growth cycle, high biomass yield and high oil content. Its biomass production capacity can reach 30 times that of terrestrial plants. The great advantage of competing for food. Microalgae has very attractive and huge development potential in the field of renewable energy. For example, microalgae has a strong oil production capacity, and its oil content can be as high as 40-80% of the dry cell weight. Therefore, microalgae is very likely to provide an effective solution to the shortage of biomass energy in my country. The development of energy microalgae resources and the development of energy microalgae biorefinery industry are not only in line with the energy ...

Claims

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Application Information

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IPC IPC(8): C12N1/22C12N1/12C12N1/20
CPCC12N1/12C12N1/20C12N1/22
Inventor 崔球刘亚君祁宽宋晓金
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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