Rhodosporidium toruloides strain and application thereof to production of torularhodin

A technology of Rhodosporidium toruloides and zymoerythrin, applied in the field of bioengineering, can solve problems such as low yield, and achieve the effect of good application prospect and economic value

Active Publication Date: 2019-03-01
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem of low yield in the production of rhodotorulin in existing red yeast, and improve the ratio and output of rhodorubin in rhodotorula strains

Method used

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  • Rhodosporidium toruloides strain and application thereof to production of torularhodin
  • Rhodosporidium toruloides strain and application thereof to production of torularhodin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Using the Agrobacterium transformation method, the wild-type Rhodosporidium toruloides np11 was mutated to obtain Rhodosporidium toruloides A1-15-BRQ

[0023] 1. Agrobacterium-mediated transformation:

[0024] (a), Agrobacterium dilution preparation

[0025] Pick a single colony of Agrobacterium engineering bacteria AGL1-HYG1 and inoculate it into 5 mL LB broth medium (containing 25 μg / mL rifampicin and 50 μg / mL kanamycin), and culture overnight at 28°C and 200 rpm / min for 15-16 hours , collect the cells, and resuspend the cells to OD with sterilized distilled water 600nm =0.6, get Agrobacterium dilution.

[0026] Among them, using Agrobacterium AGL1 competent cells (purchased from Beijing Huayueyang Biology GX9131-100), according to the method in the following literature to prepare Agrobacterium engineering bacteria AGL1-HYG1: Lin X, Wang Y, Zhang S, et al .Functional integration of multiple genes into the genome of the oleaginous yeast Rhodosporidium toruloides[J]....

Embodiment 2

[0039] Use Rhodosporidium toruloides A1-15-BRQ to produce the method in rhodotorulin, comprising the following steps:

[0040] S1. Preparation of seed liquid: Pick the Rhodosporidium toruloides A1-15-BRQ strain frozen at -80°C, that is, pick an appropriate amount of strain ice, streak it on the YEPDH plate, and culture it at 28°C for 2 days to obtain the first On the first-generation plate, pick a single colony and streak it on the YEPDH plate, and culture it at 28°C for 4 days to obtain the second-generation plate; pick a single colony on the second-generation plate and inoculate it into 50mL YEPD liquid medium (20g / L glucose , 10g / L yeast powder, 10g / L peptone, pH5.9), cultivated at 28°C and 180rpm / min for 24h to obtain seed liquid;

[0041] S2. Fermentation: Inoculate 5% of Rhodosporidium toruloides A1-15-BRQ seed liquid into 70mL liquid medium for producing rhodophyroerythrin, and cultivate in 250mL shake flask at 29°C and 180rpm / min 48h, obtain the fermented liquid that ...

Embodiment 3

[0045] Use Rhodosporidium toruloides A1-15-BRQ to produce the method in rhodotorulin, comprising the following steps:

[0046] S1. Strain activation: Pick the Rhodosporidium toruloides A1-15-BRQ strain frozen at -80°C, that is, pick an appropriate amount of strain ice, streak it on the YEPDH plate, and culture it at 28°C for 2 days to obtain the first Pick a single colony on the YEPDH plate and culture it at 28°C for 4 days to obtain the second generation plate; pick a single colony on the second generation plate and inoculate into 50mL YEPD liquid medium (20g / L glucose, 10g / L yeast powder, 10g / L peptone, pH6.2), cultured in a 250mL shake flask at 30°C and 200rpm / min for 24h to obtain Rhodosporidium toruloides A1-15-BRQ seed liquid;

[0047] S2. Fermentation: Inoculate 20% of Rhodosporidium toruloides A1-15-BRQ seed solution into 100mL liquid medium for producing rhodophyroerythrin, and cultivate in 250mL shake flask at 30°C and 190rpm / min 96h, obtain the fermented liquid tha...

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Abstract

The invention discloses a rhodosporidium toruloides strain and application thereof to production of torularhodin. According to the rhodosporidium toruloides strain, an agrobacterium-mediated transformation method is applied to leading to mutation of wild rhodosporidium toruloides np11 to obtain the rhodosporidium toruloides strain A1-15-BRQ with a preservation number of CGMCC (China General Microbiological Culture Collection Center) NO. 16638. Application of the rhodosporidium toruloides strain A1-15-BRQ to production of torularhodin comprises S1, activating bacterial seeds to obtain a seed solution; S2, inoculating the seed solution into a liquid culture medium for cultivation at 28-30 DEG C and 180-200 rpm / min for 48-96 h to obtain a fermentation solution; during application, the fermentation solution can improve the production proportion and the yield of torularhodin by 1.5-2.5 times and 4.0-9.6 times respectively compared with the wild rhodosporidium toruloides np11, thereby beingapplicable to industrial production of torularhodin and having a good application prospect and high economic values.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a strain of Rhodosporidium torulosporidium and its application in producing rhodotorulin. Background technique [0002] Rhodotorula is a class of microorganisms belonging to Basidiomycota, Rustomycetes, Basidiomycetes, Saccharomyces family, and Rhodotorula genus. Rhodotorula is multilateral budding and has the ability to produce oil and carotenoids. Rhodotorula thallus is rich in natural substances such as protein, glycogen, higher unsaturated fatty acids, astaxanthin and vitamins, so it can be used as an excellent feed and additive. Rhodosporidium toruloides is one of the common red yeasts. It can use wastewater and factory waste as carbon and nitrogen sources to produce oils and carotenoids, which can treat wastewater and fertilizers and protect the environment. [0003] Rhodotorulin is one of the carotenoids and an important precursor of vitamin A and hormones. It has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12P23/00C12R1/645
CPCC12P23/00C12R2001/645C12N1/145
Inventor 林心萍包汭琪谭明乾纪超凡梁会朋李胜杰孙泽平孙建飞
Owner DALIAN POLYTECHNIC UNIVERSITY
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