Fermentative carotenoid production

A technology of carotene and lycopene, applied in fermentation, microorganisms, sugar derivatives, etc., can solve the problem of undiscovered crtW gene functional activity combination

Inactive Publication Date: 2005-04-20
DSM IP ASSETS BV
View PDF11 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, so far, the carotenoid biosynthesis gene of the present invention has not been found to have a functionally active combination with the known crtW gene. More importantly, it is necessary to continue to explore a more optimized fermentation system for industrial production applications

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
  • Fermentative carotenoid production
  • Fermentative carotenoid production
  • Fermentative carotenoid production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Materials used and general methods

[0127] Bacterial Strains and Plasmids: Flavobacterium R 1534 wild type (ATCC 21588) was the DNA source for the cloned genes. The partial genome library of Flavobacterium R 1534 wild-type DNA was constructed into pBluescriptII+ (KS) or (SK) vector (Stratagene, La Jolla, USA), and transformed into E.coli XL-1 blue (stratagene) or JM 109.

[0128] Media and growth conditions: Transformed E. coli were grown at 37°C in Luria broth supplemented with 100 μg ampicillin (Amp) / ml for selection. Flavobacterium genus R1534 wild-type at 27 ° C, in the presence of 1% glucose, 1% tryptone (Difco Laboratories), 1% yeast extract (Difco), 0.5% MgSO 4 ·7H 2 Grow in O and 3% NaCl medium.

[0129] Colony screening: using the following primers, E. Coli transformants were screened by PCR method basically according to the method of Zon et al. [Zon et al., Biotechniques7, 696-698 (1989)]:

[0130] Primer # 7: 5′-CCTGGATGACGTGCTGGAATATTCC-3′

[0131] ...

Embodiment 2

[0142] Cloning of the Carotenoid Biosynthesis Gene of Flavobacterium R 1534

[0143] To identify and isolate DNA fragments carrying genes of the carotenoid biosynthetic pathway, the inventors used DNA fragment 46F (see Methods section) to probe the chromosomal DNA of Flavobacterium R 1534 species digested with different restriction enzymes Southern blot (Figure 2). The 2.4 kb XhoI / PstI fragment that hybridized to this probe appeared to be the most suitable fragment to start with. Genomic Flavobacterium R1534 DNA was digested with XhoI / PstI and electrophoresed on a 1% agarose gel. A region of approximately 2.4 kb on the gel was excised based on comigrating DNA markers and the DNA was isolated. The XhoI / PstI minigene library of Flavobacterium R 1534 genomic DNA was constructed into the XhoI-PstI site of pBluescriptIISK(+). Then 100 E.Coli XL1 transformants were used with primers # 7 and primers # 8 (the same primers used to obtain the 119bp fragment (46F) above), were scr...

Embodiment 3

[0175] Materials and methods for the expression of carotenoid synthases

[0176] Bacterial strains and plasmids: Use vectors pBluescriptIIKS (+) or (-) (Stratagene, La Jolla, USA) and pUC18 [Vieira and Messing, Gene 19 , 259-268 (1982); Norrander et al., Gene 26 , 101-106 (1983)] in different E. coli strains, such as XL-1 blue (Stratagene), TG1 or JM 109 for cloning. In all Bacillus subtilis transformations, strain 1012 was used. Plasmid pHP13 [Haima et al., Mol. Gen. Genet. 209 , 335-342(1987)], and plasmid p602 / 22 [LeGrice, S.F.J.in Gene Expression Technology, Go-eddel, D.V.Editot, 201-214(1990)] are capable of replicating in Bacillus subtilis and E.Coli cells Gram(+) / (-) shuttle vector. Plasmid p205 contains the VegI promoter cloned into the SmaI site of pUC18. Plasmid pXI12 is an integrating vector for constitutive expression of genes in Bacillus subtilis [Haiker et al., in 7th Int. Symposium on the Genetics of Industrial Microorganisms, June 26-July 1, 1994, Mongre...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
Login to view more

Abstract

The present invention is directed to a DNA sequence comprising one or more DNA sequences selected from the group consisting of a DNA sequence which encodes the GGPP synthase of Flavobacterium sp. R1534 (crtE), a DNA sequence which encodes the prephytoene synthase of Flavobacterium sp. R1534 (crtB), a DNA sequence which encodes the phytoene desaturase of Flavobacterium sp. R1534 (crtI), a DNA sequence which encodes the lycopene cyclase of Flavobacterium sp. R1534 (crtY) or a DNA sequence which encodes the beta -carotene hydroxylase of Flavobacterium sp. R1534 (crtZ) or DNA sequences which are substantially homolgous, vectors comprising such DNA sequences and / or a DNA sequence which encodes the beta -carotene beta 4-oxygenase of Alcaligenes strain PC-1 (crt W) or a DNA sequence which is substantially homologous, cells which are transformed by such DNA sequences and / or vectors, a process for the preparation of a desired carotenoid or a mixture of carotenoids by cultering such transformed cells and a process for the preparation of a food or feed composition. <IMAGE>

Description

[0001] This application is a divisional application of the Chinese patent application 96108114.7 filed on June 7, 1996 with the title of "Carotenoid Fermentation Production Method". technical field [0002] This application relates to the fermentative production of carotenoids. Background technique [0003] So far, more than 600 different carotenoids have been reported to be found in bacteria, yeast, fungi and plants. Organisms that can produce carotene, but there are only two of them. β-carotene and astaxanthin have been industrially produced by microorganisms and used in the food and feed industry. Beta-carotene is obtained from algae, while astaxanthin is produced by the Pfaffia strain produced using traditional mutagenesis techniques. However, Pfaffia fermentation has the disadvantage of long fermentation period, and recovery from algae is very troublesome. Therefore, people hope to develop various production systems with good industrial application value, for example...

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
Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/16A23L1/30A23L33/155C07H21/04C07K14/195C12N1/21C12N15/09C12N15/52C12N15/53C12P7/04C12P23/00C12R1/05C12R1/125C12R1/19C12R1/20
CPCC07K14/195C12N15/52C12P23/00
Inventor 汉斯-彼得·霍曼卢斯·帕萨蒙特斯米歇尔·特西尔阿道弗斯·冯卢恩
Owner DSM IP ASSETS BV
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