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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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 li

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

Examples

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Example Embodiment

[0050] Example 1: Construction of cellulase preparation based on Clostridium thermocellum cellosome by indirect connection method

[0051] Through seamless cloning, the tdk expression cassette (containing the promoter of the gapDH gene) and the pyrF expression cassette (containing the pyrF 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 the tdk and pyrF expression cassette, and EagI and MluI restriction sites were added downstream of pyrF for the cloning of homologous arm fragments, thereby constructing pHK-HR Plasmid.

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

Example Embodiment

[0058] Example 2: Construction of a cellulase preparation based on Clostridium thermocellum cellulosomes through indirect connection

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

Example Embodiment

[0060] Example 3: Construction of cellulase preparations based on Clostridium thermocellum cellulosomes by means of direct connection

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

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

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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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