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Method for increasing yield of monascus pigment

A technology of monascus pigment and yield, which is applied in the field of industrial microorganisms, can solve the problems of difficulty in directional accumulation of alcohol-soluble monascus pigment and low yield of monascus pigment

Active Publication Date: 2019-02-15
JIANGXI SCI & TECH NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the technical defects of the prior art, and provides a method for increasing the yield of monascus pigment, so as to solve the technical problem of low yield when conventionally cultivating Monascus red to produce monascus pigment in the prior art
[0007] Another technical problem to be solved by the present invention is that it is difficult to directionally accumulate alcohol-soluble monascus pigments when conventionally cultivating Monascus ruberus to produce monascus pigments in the prior art

Method used

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  • Method for increasing yield of monascus pigment
  • Method for increasing yield of monascus pigment
  • Method for increasing yield of monascus pigment

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Experimental program
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Effect test

Embodiment 1

[0053] 1. Construction of the binary plasmid knockout vector pHph0380.

[0054] According to the commercial plant binary plasmid pCambia0380 is the original vector. Design a pair of oligonucleotide sequence F&R, the sequence was synthesized by Shanghai Shenggong Biological Engineering Co., Ltd. This sequence contains the following restriction endonuclease sites (Hind III, Kpn I, Sac I, Pac I, Pme I, Xho I, Xba I, Bgl II) in sequence. The binary plasmid vector pCambia0380 was digested with restriction endonucleases Hind III and Bgl II. The oligonucleotide sequence was ligated with the digestion vector pCambia0380 by T4DNA ligase to obtain the binary plasmid vector pCambia0380G.

[0055] Using the plasmid pMD19-PgpdA-hph-TtrpC preserved in the laboratory as a template, the hph expression cassette was amplified by primers PgpdA-Sac I-F&TtrpC-Xho I-R. The restriction endonucleases Sac I and Xho I were used to simultaneously digest the hph expression cassette fragment and the binary ...

Embodiment 2

[0108] 1. Construction of binary plasmid knockout vector pHph0380

[0109] According to the commercial plant binary plasmid pCambia0380 is the original vector. Design a pair of oligonucleotide sequence F&R, which contains the following restriction endonuclease sites (Hind III, Kpn I, Sac I, Pac I, Pme I, Xho I, XbaI, Bgl II) in sequence. The binary plasmid vector pCambia0380 was digested with restriction endonucleases Hind III and Bgl II. The oligonucleotide sequence was ligated with the vector by T4DNA ligase to obtain the binary plasmid vector pCambia0380G.

[0110] Using the plasmid pMD19-PgpdA-hph-TtrpC preserved in the laboratory as a template, the hph expression cassette fragment was amplified by primers PgpdA-Sac I-F&TtrpC-Xho I-R. The restriction endonucleases Sac I and Xho I were used to simultaneously digest the promoter fragment and the binary plasmid vector pCambia0380G. The oligonucleotide sequence was ligated with the vector by T4DNA ligase to obtain the binary pla...

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Abstract

The invention provides a method for increasing yield of monascus pigment. According to the technical scheme, starting from a plant dual-element plasmid pCambia0380 vector, a dual-element plasmid vector pHph0380 suitable for knockout of flamentous fungus genes is modified and constructed. On the basis, upstream and downstream homologous arm fragments of glucose transporter gltp1 genes are cloned from monascus ruber CICC41233 and connected with the plasmid vector pHph0380, the fragments are transformed into parent monascus ruber through agrobacterium tumefaciens EHA105 mediation, and a recombinant strain is constructed. Monascus pigment fermentation results prove that the recombinant strain constructed with the method has monascus pigment accumulation time advanced significantly, the alcohol-soluble pigment yield is obviously increased, and when fermentation is performed for 144 h, the alcohol-soluble pigment yield is 74% higher than that of parent strains. The fermentation time of monascus for producing monascus pigment is advanced by use of a gene engineering means, the utilization rate of starch is increased, and the yield of the monascus pigment is increased.

Description

Technical field [0001] The present invention relates to the technical field of industrial microorganisms, and further relates to genetic engineering technology and mold fermentation technology, in particular to a method for increasing the production of monascus pigment. Background technique [0002] Monascus pigment is a natural pigment fermented from rice by the microorganism Monasus spp., and has a history of more than a thousand years in my country. As a food additive, monascus pigment is widely used in food processing and cosmetics manufacturing and other fields; because it also has a wide range of biological activities such as regulating blood lipids, lowering blood pressure, preventing vascular sclerosis, anti-diabetics, inhibiting obesity, anti-inflammatory, anti-allergic, and preventing Peroxidation, anti-cancer, anti-bacterial, anti-fungal, etc., its application in the development of probiotics and health care products and medical fields has also received more and more a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P1/02C12N15/80C12N1/15C12R1/645
CPCC07K14/37C12N15/80C12P1/02C12N1/145C12R2001/645
Inventor 龙传南曾斌陶琴琴刘心怡刘梦梦彭玲程芳婷王淑琴吾蔚蔚
Owner JIANGXI SCI & TECH NORMAL UNIV
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