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Method for preparing astaxanthin by adopting lignocellulose

A lignocellulose and astaxanthin technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as insufficient efficiency and cost, and difficulty in large-scale application of lignocellulose bioconversion, and achieve The effect of reducing chemical use, lowering the cost of enzymes, and saving water

Active Publication Date: 2018-12-11
青岛中科潮生生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the biotransformation of lignocellulose generally uses cellulase derived from fungi, which has insufficient efficiency and cost. It is difficult to achieve large-scale application of lignocellulose biotransformation based on cellulase preparations derived from fungi.

Method used

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  • Method for preparing astaxanthin by adopting lignocellulose
  • Method for preparing astaxanthin by adopting lignocellulose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: By the method of indirect connection, construct the cellulase preparation based on Clostridium thermocellum cellulite

[0050] 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.

[0051] The cellulase Cel9K (exocellulase, encoded by the nucleic acid sequence 2113813 to 2111293 in the genome CP002416.1) in the celluloso...

Embodiment 2

[0057] Embodiment 2: By the method of indirect connection, construct the cellulase preparation based on Clostridium thermocellum cellulite

[0058] The difference from Example 1 is that the polypeptide fragment II or polypeptide fragment I was 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

[0059] Embodiment 3: By the method of direct connection, construct the cellulase preparation based on Clostridium thermocellum cellulosome

[0060] 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 (SEQ ID 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.

[0061] 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...

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Abstract

The invention provides a technology for producing astaxanthin through efficient saccharifying of lignocellulose type agricultural and forest wastes, aiming at the problems in the aspects of producingthe astaxanthin by fermenting through phaffia rhodozyma and carrying out enzymolysis on lignocellulose. The technology comprises the steps of pre-treating, saccharifying, carrying out phaffia rhodozyma fermentation, extracting the astaxanthin and the like. According to the technology provided by the invention, a lignocellulose biomass is used as a raw material, so that on one hand, the enzyme utilization cost of a saccharifying phase and a production cost of microorganism fermentation are remarkably improved; on the other hand, the comprehensive utilization problems of the agricultural and forest wastes are also solved. By combining biological saccharifying of the lignocellulose and a high-density fermentation phase of the phaffia rhodozyma, ash obtained by saccharifying can be further used as a solid matrix of yeast fermentation and the fermentation density is improved. In the technology provided by the invention, the ratio of the astaxanthin to the dry weight of thalli can reach 2.6percent and the biomass of yeast thalli can reach 65g / L. The purity of the obtained refined astaxanthin is greater than 90 percent through technologies including molecular distillation and the like.

Description

technical field [0001] The invention relates to the field of microbial fermentation engineering, in particular to a method for producing astaxanthin by using lignocellulose raw materials. Background technique [0002] Astaxanthin, also known as astaxanthin, lobster shell pigment, chemical name: 3,3′-dihydroxy-4,4′-diketone β-carotene, pigment Aj067-69CAS No:472-61 -7, molecular formula C40H52O4, molecular weight 596.86. Astaxanthin is a carotenoid with the strongest antioxidant properties, which can effectively remove oxygen free radicals in cells, enhance cell regeneration, maintain body balance and reduce the accumulation of aging cells, thereby protecting skin health and promoting hair growth , anti-aging, relieve sports fatigue, enhance vitality. In addition, astaxanthin has the functions of improving immunity, preventing tumors, cardiovascular diseases and eye diseases, so it has broad application prospects in the fields of food, medicine and cosmetics. [0003] Alth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P23/00C12P19/14C12P19/02C12R1/645
CPCC12P19/02C12P19/14C12P23/00C12P2201/00C12P2203/00
Inventor 崔球刘亚君宋晓金
Owner 青岛中科潮生生物技术有限公司
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