Method for preparing squalene by adopting lignocellulose

A technology of lignocellulose and squalene, which is applied in chemical instruments and methods, purification/separation of hydrocarbons, waste fuels, etc., can solve the problems of inability to interact with cellulosic bodies, complicated processes, and reduced synergy, etc. Comprehensive utilization problems, wide source of raw materials, and the effect of strong market competitiveness

Active Publication Date: 2018-11-23
青岛中科潮生生物技术有限公司
View PDF7 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the activity and stability of exogenously added proteins will decrease with the progress of the saccharification process, the consumption of enzymes requires an increase in the amount or frequency of enzyme addition
In addition, these non-cellulosomal proteins are added to the saccharification system as free enzymes, which cannot interact with cellulosomes, and will significantly reduce the s...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for preparing squalene by adopting lignocellulose
  • Method for preparing squalene by adopting lignocellulose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: By the method of indirect connection, construct the cellulase preparation based on Clostridium thermocellum cellulite

[0049] 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.

[0050] The cellulase Cel9K (exocellulase, encoded by the nucleic acid sequence 2113813 to 2111293 in the genome CP002416.1) in the celluloso...

Embodiment 2

[0056] Embodiment 2: By the method of indirect connection, construct the cellulase preparation based on Clostridium thermocellum cellulite

[0057] 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

[0058] Embodiment 3: By the method of direct connection, construct the cellulase preparation based on Clostridium thermocellum cellulosome

[0059] 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.

[0060] 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...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a process for preparing squalene by taking lignocellulosic agricultural and forestry wastes as raw materials, which aims at problems in the aspect of the production of squaleneby microbial fermentation in the prior art. The process comprises the steps of pre-treatment, saccharification, yeast-like fungi fermentation and squalene extraction and the like. According to the process provided by the invention, lignocellulose biological saccharification is combined with yeast-like fungi high-density fermentation; by adopting lignocellulose biomass as raw material, on the one hand, the cost of enzyme usage and the production cost of squalene microbial fermentation in a saccharification stage are significantly reduced; on the other hand, the problem of comprehensive utilization of the agricultural and forestry wastes is solved. In addition, medium and fermentation medium in the lignocellulose saccharification stage can be recycled, water can be saved and usage of chemicals can be reduced, and has significant effect on reducing the discharge of wastewater and reducing the cost. In the process provided by the invention, the squalene yield can reach 13 g/L, accounting for 5% of the dry weight of thallus, and the purity of refined squalene is higher than 93%.

Description

technical field [0001] The invention relates to the field of microbial fermentation engineering, in particular to a method for producing squalene by using lignocellulose raw materials. Background technique [0002] Squalene is a non-toxic marine biologically active substance with the effect of preventing and curing diseases. Its chemical name is 2,6,10,15,19,23-hexamethyl-2,6,10,14 ,18,22-Tetracosahexaene has many physiological functions such as increasing the activity of superoxide dismutase in the body, enhancing the body's immunity, improving sexual function, anti-aging, anti-fatigue, anti-tumor, etc. It is widely used in medicine, Beauty, cosmetics, food and other fields. [0003] Currently, the main source of squalene is fish oil, especially shark liver oil. However, with the depletion of fishery resources and the improvement of relevant regulations, people are trying to find other sources of squalene; such as from olive oil, palm oil and other oil crops, or from amar...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12P5/02C12P19/14C12P19/02C07C7/00C07C7/04C07C7/10C07C11/21
CPCC07C7/005C07C7/04C07C7/10C12P5/026C12P19/02C12P19/14C07C11/21Y02E50/30
Inventor 崔球刘亚君宋晓金
Owner 青岛中科潮生生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap